Search results for: recombinant E. coli

Authors: Jou-An Chen, Julio Castillo, Errol Duncan Cason, Gabre Kemp, Leana Esterhuizen, Angel Valverde Portal, Esta Van Heerden

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Emerging contaminants (EC) such as pharmaceutical and personal care products have been accumulating for years in water bodies all over the world. However, very little is known about the occurrences, levels, and effects of ECs in South African water resources. This study provides an initial assessment of the distribution of eight ECs (Acetaminophen, Atrazine, Terbuthlyazine, Carbamazepine, Phenyton, Sulfmethoxazole, Nevirapine and Fluconozole) in fifteen water sources from the Free State and Easter Cape provinces of South Africa. Overall, the physiochemical conditions were different in surface and groundwater samples, with concentrations of several elements such as B, Ca, Mg, Na, NO3, and TDS been statistically higher in groundwater. In contrast, ECs levels, quantified at ng/mL using the LC/MS/ESI, were much lower in groundwater samples. The ECs with higher contamination levels were Carbamazepine, Sulfmethoxazole, Nevirapine, and Terbuthlyazine, while the most widespread were Sulfmethoxazole and Fluconozole, detected in all surface and groundwater samples. Fecal and E. coli tests indicated that surface water was more contaminated than groundwater. Microbial communities, assessed using NGS, were dominated by the phyla Proteobacteria and Bacteroidetes, in both surface and groundwater. Actinobacteria, Planctomycetes, and Cyanobacteria, were more dominant in surface water, while Verrucomicrobia were overrepresented in groundwater. In conclusion, ECs contamination is closely associated with human activities (human wastes). The microbial diversity identified can suggest possible biodegradation processes.

Keywords: emerging contaminants, EC, personal care products, pharmaceuticals, natural attenuation process

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Authors: Tesfay Gebremicheal Reda, K. Samatha, Paul Douglas Sanasi, D. Parajuli

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Nanoparticle technology is fast progressing and is being employed in innumerable medical applications. At this time, the public's health is seriously threatened by the rise of bacterial strains resistant to several medications. Metal nanoparticles are a potential alternate approach for tackling this global concern, and this is the main focus of this study. The citrate precursor sol-gel synthesis method was used to synthesize the Niₓ Co₁₋ₓ Fe₂ O₄, (where x = 0.0:0.2:1.0) nanoparticle. XRD identified the development of the cubic crystal structure to have a preferential orientation along (311), and the average particle size was found to be 29-38 nm. The average crystallizes assessed with ImageJ software and origin 22 of the SEM are nearly identical to the XRD results. In the created NCF NPs, the FT-IR spectroscopy reveals structural examinations and the redistribution of cations between octahedral (505-428 cm⁻¹) and tetrahedral (653-603 cm⁻¹) locales. As the Co²⁺ cation is substituted with Ni²⁺, the coercive fields HC decrease from 2384 Oe to 241.93 Oe. Band gap energy rises as Ni concentration increases, which may be attributed to the fact that the ionic radii of Ni²⁺ ions are smaller than that of Co²⁺ ions, which results in a strong electrostatic interaction. On the contrary, except at x = 0.4, the dielectric constant decreases as the nickel concentration increases. According to the findings of this research work, nanoparticles are composed of Ni₀.₄ Co₀.₆ Fe₂ O₄ have demonstrated a promising value against S. aureus and E. coli, and it suggests a proposed model for their potential use as a source of antibacterial agent.

Keywords: antimicrobial, band gap, citrate precursor, dielectric, nanoparticle

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Authors: Sifi Ibrahim, Benaddou Fatima Zohra, Yousfi Mohamed

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The Marrubium deserti and Ammodaucus leucotrichus L. an Algerian endemic species, has several applications in traditional medicine for example as a remedy for asthma and diabetes, and was found to have antibacterial properties. In this work, an antioxidant and antimicrobial activities was performed on phenolic extracts of Marrubium deserti, Ammodaucus leucotrichus plants. The yield of methanol maceration of these plants is 12.4% and 20.4% respectively. The content of total polyphenols, flavonoids and anthocyanin in methanolic extracts, are varied between 19.52±1.88 and 59.24±3.45 mg/g gallic acid equivalent, and 2.08±0.29 to 1.46±0.39 mg/g quercetin equivalent, and 0.395 to 1.934µmol/g respectively. The total chlorophylls and carotenoids were be ranged from 0.149±0.20 to 1.537±0.20 g/ml and 1.537±0.20 to 0.149 ± 0.20 g/ml, respectively. According to DPPH and FRAP test, the values of EC50 was shows a higher activity of Marrubium deserti than Ammodaucus leucotrichus with EC50 values (DPPH) were 34.53±0.71 μg/mL and 258.60±15.67 mg/ml respectively. The TEAC values of FRAP test was a highly superior for Marrubium deserti 209.66±0.26 mg Equivalent Trolox/g dry residue than Ammodaucus leucotrichus 45.88±2.93 mg Trolox Equivalent/g dry residue. The antimicrobial activity against nine strains of bacteria (Staphylococcus aureus(+), Staphylococcus aureus (-), Bacillus cereus, Enterococcus faecalis, Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae and Salmonella typhi), was showed that the tested extracts are a significant antibacterial activity with inhibition zones ranging from 10 to 50 mm. the value of CMI were ranging from 0.89 to 14.29 mg/ml.

Keywords: phenolic extract, antioxidant activity, antimicrobial activity, Marrubium deserti, Ammodaucus leucotrichus

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Authors: Aulanni’am Aulanni’am

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Incidence of Diabetes Mellitus (DM) progressively increasing it became a serious problem in Indonesia and it is a disease that government is priority to be addressed. The longer a person is suffering from diabetes the more likely to develop complications particularly diabetic patients who are not well maintained. Therefore, Incidence of Diabetes Mellitus needs to be done in the early diagnosis of pre-phase of the disease. In this pre-phase disease, already happening destruction of pancreatic beta cells and declining in beta cell function and the sign autoimmunity reactions associated with beta cell destruction. Type 1 DM is a multifactorial disease triggered by genetic and environmental factors, which leads to the destruction of pancreatic beta cells. Early marker of "beta cell autoreactivity" is the synthesis of autoantibodies against 65-kDa protein, which can be a molecule that can be detected early in the disease pathomechanism. The importance of early diagnosis of diabetic patients held in the phase of pre-disease is to determine the progression towards the onset of pancreatic beta cell destruction and take precautions. However, the price for this examination is very expensive ($ 150/ test), the anti-GAD65 abs examination cannot be carried out routinely in most or even in all laboratories in Indonesia. Therefore, production-based Rapid Test Recombinant Human Protein GAD65 with "Reverse Flow Immunchromatography Technique" in Indonesia is believed to reduce costs and improve the quality of care of patients with diabetes in Indonesia. Rapid Test Product innovation is very simple and suitable for screening and routine inspection of GAD65 autoantibodies. In the blood serum of patients with diabetes caused by autoimmunity, autoantibody-GAD65 is a major serologic marker to detect autoimmune reaction because their concentration level of stability.GAD65 autoantibodies can be found 10 years before clinical symptoms of diabetes. Early diagnosis is more focused to detect the presence autontibodi-GAD65 given specification and high sensitivity. Autoantibodies- GAD65 that circulates in the blood is a major indicator of the destruction of the islet cells of the pancreas. Results of research in collaboration with Biofarma has produced GAD65 autoantibodies based Rapid Test had conducted the soft launch of products and has been tested with the results of a sensitivity of 100 percent and a specificity between 90 and 96% compared with the gold standard (import product) which worked based on ELISA method.

Keywords: diabetes mellitus, GAD65 autoantibodies, rapid test, sensitivity, specificity

Procedia PDF Downloads 233

Authors: S. Mantzoukis, M. Gerasimou, P. Christodoulou, N. Varsamis, G. Kolliopoulou, N. Zotos

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Purpose: Patients in Intensive Care Units (ICU) are exposed to a different spectrum of microorganisms relative to the hospital. Due to the fact that the majority of these patients are intubated, bronchial secretions should be examined. Material and Method: Bronchial secretions should be taken with care so as not to be mixed with sputum or saliva. The bronchial secretions are placed in a sterile container and then inoculated into blood, Mac Conkey No2, Chocolate, Mueller Hinton, Chapman and Saboureaud agar. After this period, if any number of microbial colonies are detected, gram staining is performed and then the isolated organisms are identified by biochemical techniques in the automated Microscan system (Siemens) followed by a sensitivity test in the same system using the minimum inhibitory concentration MIC technique. The sensitivity test is verified by a Kirby Bauer test. Results: In 2017 the Laboratory of Microbiology received 365 samples of bronchial secretions from the Intensive Care Unit. 237 were found positive. S. epidermidis was identified in 1 specimen, A. baumannii in 60, K. pneumoniae in 42, P. aeruginosa in 50, C. albicans in 40, P. mirabilis in 4, E. coli in 4, S. maltophilia in 6, S. marcescens in 6, S. aureus in 12, S. pneumoniae in 1, S. haemolyticus in 4, P. fluorescens in 1, E. aerogenes in 1, E. cloacae in 5. Conclusions: The majority of ICU patients appear to be a fertile ground for the development of infections. The nature of the findings suggests that a significant part of the bacteria found comes from the unit (nosocomial infection).

Keywords: bronchial secretions, cultures, infections, intensive care units

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Authors: Arfan Ali, Idrees Ahmad Nasir

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Potato (Solanum tuberosum) is an important cash crop and popular vegetable in Pakistan and throughout the world. Cold storage of potatoes accelerates the conversion of starch into reduced sugars (glucose and fructose). This process causes dry mass and bitter taste in the potatoes that are not acceptable to end consumers. In the current study, the phosphofructokinase B gene was cloned into the pET-30 vector for protein expression and the pCambia-1301 vector for plant expression. Amplification of a 930bp product from an E. coli strain determined the successful isolation of the phosphofructokinase B gene. Restriction digestion using NcoI and BglII along with the amplification of the 930bp product using gene specific primers confirmed the successful cloning of the PfkB gene in both vectors. The protein was expressed as a His-PfkB fusion protein. Western blot analysis confirmed the presence of the 35 Kda PfkB protein when hybridized with anti-His antibodies. The construct Fani-01 was evaluated transiently using a histochemical gus assay. The appearance of blue color in the agroinfiltrated area of potato leaves confirmed the successful expression of construct Fani-01. Further, the area displaying gus expression was evaluated for PfkB expression using ELISA. Moreover, PfkB gene expression evaluated through transient expression determined successful gene expression and highlighted its potential utilization for stable expression in potato to reduce sweetening due to long-term storage.

Keywords: potato, Solanum tuberosum, transformation, PfkB, anti-sweetening

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Authors: Adrian Ionut Nicoara, Denisa Ionela Ene, Alina Maria Holban, Cristina Busuioc

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At present, the development of materials with biomedical applications is a domain of interest that will produce a full series of benefits in engineering and medicine. In this sense, it is required to use a natural material, and this paper is focused on the development of a composite material based on bacterial cellulose – hydroxyapatite and silver nanoparticles with applications in hard tissue. Bacterial cellulose own features like biocompatibility, non-toxicity character and flexibility. Moreover, the bacterial cellulose can be conjugated with different forms of active silver to possess antimicrobial activity. Hydroxyapatite is well known that can mimic at a significant level the activity of the initial bone. The material was synthesized by using an ultrasound probe and finally characterized by several methods. Thereby, the morphological properties were analyzed by using Scanning Electron Microscopy (SEM) and Transmission Electron Microscopy (TEM). Because the synthesized material has medical application in restore the tissue and to fight against microbial invasion, the samples were tested from the biological point of view by evaluating the biodegradability in phosphate-buffered saline (PBS) and simulated body fluid (SBF) and moreover the antimicrobial effect was performed on Gram-positive bacterium Staphylococcus aureus, Gram-negative bacterium Escherichia coli, and fungi Candida albicans. The results reveal that the obtained material has specific characteristics for bone regeneration.

Keywords: bacterial cellulose, biomaterials, hydroxyapatite, scaffolds materials

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Authors: Rafael Estrada, Cristian Ruiz Rueda

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Carbapenems are last-resort antibiotics used in clinical settings to treat antibiotic-resistant bacterial infections. Thus, the emergence and spread of resistance to carbapenems is a major public health concern. Here, we have studied a carbapenem-resistant Aeromonas veronii strain previously isolated from a water sample from Sam Simeon Creek (Hearst San Simeon State Park, CA). Analysis of this isolate using disk-diffusion, CarbaNP, eCIM and mCIM assays revealed that it was resistant to amoxicillin-clavulanic acid and all carbapenems tested and that this isolate produced a potentially novel carbapenemase of the Metallo-β-lactamase family. Whole genome sequencing analysis revealed that this A. veronii isolate carries a novel variant of the blacₚₕₐ class β-carbapenemase gene that was closely related to the blacₚₕₐ₇ gene of Aeromonas jandaei. This isolate also carried a novel variant of the blaₒₓₐ class D carbapenemase gene that was most closely related to the blaₒₓₐ-₉₁₂ gene found in other Aeromonas veronii isolates. Finally, we also identified a novel class C β-lactamase gene moderately related to the blaFₒₓ-₁₇ gene of Providencia stuartii and other blaFₒₓ variants identified in Klebsiella pneumoniae, Escherichia coli and other Enterobacteriaceae. Overall, our findings reveal that environmental isolates are an important reservoir of multiple carbapenemases and other β-lactamases of clinical significance.

Keywords: β-lactamases, carbapenem, antibiotic-resistant, aeromonas veronii

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Authors: Hafsa Tahir, Sanaullah Iqbal

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Glactooligosaccharide (GOS) is a type of prebiotic which is mainly found in human milk. GOS belongs to those bacteria which stimulates the growth of beneficial bacteria in human intestines. The aim of the present study was to develop a value-added product by producing prebiotic (GOS) in buttermilk through trans galactosylation. Buttermilk is considered as an industrial waste which is discarded after the production of butter and cream. It contains protein, minerals, vitamins and a smaller amount of fat. Raw milk was pasteurized at 100º C for butter production and then trans galactosylation process was induced in the butter milk thus obtained to produce prebiotic GOS. Results showed that the enzyme (which was obtained from bacterial strain of Esecrshia coli and has a gene of Lactobacillus reuteri L103) concentration between 400-600µl/5ml can produce GOS in 30 minutes. Chemical analysis and sensory evaluation of plain and GOS containing buttermilk showed no remarkable difference in their composition. Furthermore, the shelf-life study showed that there was non-significant (P>0.05) difference in glass and pouch packaging of buttermilk. Buttermilk in pouch packaging maintained its stability for 6 days without the addition of preservatives. Therefore it is recommended that GOS enriched buttermilk which is generally considered as a processing waste in dairy manufacturing can be turned into a cost-effective nutritional functional food product. This will not only enhance the production efficiency of butter processing but also will create a new market opportunity for dairy manufacturers all over the world.

Keywords: buttermilk, galactooligosaccharide, shelf Life, transgalactosylation

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Authors: K. P. Ganesh, T. Gomathi, L. Arul Pragasan

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Degradation of lake water quality is one of the serious environmental threats for the last few decades, particularly, the lakes situated in and around urban and industrial areas. The present study aimed to analyze the physicochemical and biological parameters, and metal elements to determine the water quality of Krishnampathi, Ukkadam, Kurichi, Sulur and Singanallur Lakes. Of the 23 physicochemical parameters analyzed in the five lakes, except TDS, Chloride and Total hardness values all the 20 parameters were found within the prescribed limit as recommended by World Health Organization (WHO) and Bureau of Indian Standards (BIS). In case of biological parameter, both Total Coliform and Fecal Coliform bacteria (Escherichia coli) were identified. This indicates the contamination of lakes by fecal matter, and warns of potential of disease causing by viruses, bacteria and other organisms. Among the twelve metal elements (Al, Cr, Mn, Fe, Ni, Cu, Zn, As, Se, Mo, Cd and Pb) determined by inductively coupled plasma-mass spectroscopy, except Cd (for all lakes), and Pb (for Ukkadam, Kurichi, Sulur & Singanallur), all the elements were found above the prescribed limits of BIS. The results of the present study revealed that all the five major lakes of Coimbatore were contaminated. It is recommended that proper implementation of the new wetland waste management system and monitoring of water quality be of the urgent need to sustain the water bodies for future generations.

Keywords: heavy metals, inductively coupled plasma-mass spectroscopy, physicochemical and biological parameters, water quality

Procedia PDF Downloads 151

Authors: Alvine Sandrine Ndinchout, D. P. Chattopadhyay, Moundipa Fewou Paul, Nyegue Maximilienne Ascension, Varinder Kaur, Sukhraj Kaur, B. H. Patel

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Dacryodes macrophylla is a species of the Burseraceae family that is widespread in Cameroon, Equatorial Guinea, and Gabon. The only part of D. macrophylla known to use is the pulp contained in the fruit. This very juicy pulp is consumed directly and used in making juices. During consumption, these fruit leaves a dark blackish colour on fingers and garment. This observation means that D. macrophylla fruits must be a good source of natural dye with probably good fastness properties on textile materials. But D. macrophylla has not yet been investigated with reference as a potential source of natural dye to our best knowledge. Natural dye has been extracted using water as solvent by soxhlet extraction method. The extracted color was characterized by spectroscopic studies like UV/Visible and further tested for antimicrobial activity against gram-negative (Vibrio cholerae, Escherichia coli, Salmonella enterica serotype Typhi, Shigella flexneri) and gram-positive (Listeria monocytogenes, Staphylococcus aureus) bacteria. It was observed that the water extract of D. macrophylla showed antimicrobial activities against S. enterica. The results of fastness properties of the dyed fabrics were fair to good. Taken together, these results indicate that D. macrophylla can be used as natural dye not only in textile but also in other domains like food coloring.

Keywords: antimicrobial activity, natural dye, silk, wash fastness, wool

Procedia PDF Downloads 142

Authors: Amina Ben Abla, Sylvie Changotade, Geraldine Rohman, Guilhem Boeuf, Cyrine Dridi, Ahmed Elmarjou, Florence Dufour, Didier Lutomski, Abdellatif Elm’semi

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Understanding cell adhesion and interaction with the extracellular matrix is essential for biomedical and biotechnological applications, including the development of biomaterials. In recent years, numerous biomaterials have emerged and were used in the field of tissue engineering. Nevertheless, the lack of interaction of biomaterials with cells still limits their bio-integration. Thus, the design of bioactive biomaterials to improve cell attachment and proliferation is of growing interest. In this study, bio-functionalized material was developed combining a synthetic polymer scaffold surface with selected domains of type III human fibronectin (FNIII-DOM) to promote cell adhesion and proliferation. Bioadhesive ligand includes cell-binding domains of human fibronectin, a major ECM protein that interacts with a variety of integrins cell-surface receptors, and ECM proteins through specific binding domains were engineered. FNIII-DOM was produced in bacterial system E. coli in 5L fermentor with a high yield level reaching 20mg/L. Bioactivity of the produced fragment was validated by studying cellular adhesion of human cells. The adsorption and immobilization of FNIII-DOM onto the polymer scaffold were evaluated in order to develop an innovative biomaterial.

Keywords: biomaterials, cellular adhesion, fibronectin, tissue engineering

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Authors: Hayet El Kolli, Hocine Laouer

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Bunium in the world comprises about 50 to 100 species, mostly distributed in: Algeria, Italy, Pakistan, Iran, and South Africa. Bunium species have several uses like: Bunium persicum which is commonly used as antispasmodic, carminative, anti-obesity and lactogage. This plant have been widely used as an additive in food stuff such as in bread cooking, rice and yoghurt for its carminative, anti-dyspepsia and antispasmodic effect. The B. paucifolium oil has a wide spectrum of action against moulds, yeast and bacteria. The chemical compositions of Bunium incrassatum and Bunium alpinum essential oils were carry out by GC and GC/MS. Therefore, antibacterial activity of two oils was investigated by disk diffusion method against Pseudomonas aeruginosa ATCC 27853, Escherichia coli ATCC 25922, Salmonella typhimurium ATCC 1331, Staphylococcus aureus ATCC 25923, Klebsiella pneumoniae ATCC 700603, Bacillus cereus ATCC 10876, Enterococcus faecalis ATCC 49452, Lysteria monocytogenes ATCC 15313, Citrobacter freundii ATCC 8090, Proteus mirabilis ATCC 35659. A moderate antibacterial activity was found. In conclusion, it is found that essential oils of the two species are rich in sesquiterpens and other oxygenated compounds. These compounds have been reported to show bactericidal activity and the presence of phenolic compounds makes them useful antioxidants so that results confirm some ethnopharmacologique applications of these two oils of Bunium.

Keywords: Bunium alpinum, Bunium incrassatum, apiaceae, essential oil, sesquiterpens, phenols, antibacterial, antioxidant activities

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Authors: Elmineh Tsegahun Gedif

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Zinc oxide nanoparticles (ZnO NPs) have attracted huge attention due to catalytic, optical, photonic, and antibacterial activity. Zinc oxide nanoparticles were successfully synthesized via a fast, non-toxic, cost-effective, and eco-friendly method by biologically reducing Zn(NO3)2.6H2O solution with Neem (Azadirachta indica) leaf extract under optimum conditions (pH = 9). The presence of active flavonoids, phenolic groups, alkaloids, terpenoids, and tannins, which were in the biomass of the Neem leaf extract before and after reduction, was identified using qualitative screening methods (observing the color changes) and FT-IR Spectroscopy. The formation of ZnO NPs was visually indicated by the color changes from colorless to light yellow color. Biosynthesized nanoparticles were also characterized by UV-visible, FT-IR, and XRD spectroscopies. The reduction process was simple and convenient to handle and was monitored by UV-visible spectroscopy that showed surface plasmon resonance (SPR) of the ZnO NPs at 321 nm. This result clearly revealed the formation of ZnO NPs. X-ray diffraction was used to investigate the crystal structure. The average particle size of ZnO powder and around 20 nm using the line width of the plane, and the refraction peak using Scherrer’s equation. The synthesized zinc oxide nanoparticles were evaluated for antimicrobial activities against Gram-positive and Gram-negative bacteria. Zinc nanoparticles exhibited the maximum zone of inhibition against Escherichia coli (15 mm), while the least activity was seen against Staphylococcus aureus.

Keywords: antimicrobial activity, azadirachta indica, green synthesis, ZnO NPs

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Authors: Muhammad Arshad, Ghulam Hussain Bhatti, Abdul Qayyum

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Zinc-doped silica oxide nanoparticles having size 7.93nm were synthesized by the deposition precipitation method by using different solvents (acetonitrile, n-hexane, isoamylalchol). Biological potential such as antibacterial activities against Bacillussubtilusand Escherichia coli, and antifungal activities against Candida parapsilosis and Aspergilusniger were also investigated by Disc diffusion method. Different characterizations techniques including Fournier Transmission Infrared Spectroscopy (FT-IR), X-ray diffraction (XRD), Scanning Electron Microscope (SEM), Transmission Electron Microscope (TEM), Thermo-gravimeteric Analysis (TGA), Atomic forced microscopy (AFM), and Dynamic Light Scattering (DLS) were used. FT-IR characterization confirmed the presence of metal oxide bond (SiO2) while XRD showed the hexagonal structure. SEM and TEM characterization showed the morphology of nanoparticles. AFM study showed good particle size distribution as depicted by a histogram. DLS study showed the gradual decease in the size of nanoparticles from 24.86nm to 13.24 nm. Highest antibacterial activities revealed by acetonitrile solvents (6%and 4.5%) followed by isoamylalchol (3% and 2.4%) while n-hexane solvent showed the lowest activity (2%and 1%) respectively. Higher antifungal activities exhibited by n-hexane (0.34 % and 0.43%) followed by isoamylalchol (0.27% and 0.19%) solvent while acetonitrile (0.21% and 0.17%) showed least activity respectively. Statistical analysis by using one-way ANOVA also indicated the significant results of both biological activities.

Keywords: nanoparticles, precipitation methods, antibacterial, antifungal, characterizations

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Authors: Justin Ntokamunda Kadima, Christian Ahadi Irenge, Patient Birindwa Mulashe, Félicien Mushagalusa Kasali, Patient Wimba

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Background: Bacterial strains carrying multidrug resistance traits are gaining ground worldwide, especially in countries with limited resources. This study aimed to evaluate the spreading of multidrug-resistant bacteria strains in Bukavu city hospitals in the Democratic Republic of Congo. Methods: We analyzed 758 antibiogram data recorded in files of patients consulted between January 2016 and December 2017 at three reference hospitals selected as sentinel sites, namely the Panzi General Reference Hospital (HGP), BIO -PHARM hospital (HBP), and Saint Luc Clinic (CSL). Results: Of 758 isolates tested, the laboratories identified 12 bacterial strains in 712 isolates, of which 223 (29.42%) presented MDR profile, including Escherichia coli (11.48%), Klebsiella pneumonia (6.07%), Enterobacter (5.8%), Staphylococcus aureus and coagulase-negative Staphylococci (1.58%), Proteus mirabilis (1.85%), Salmonella enterica (1.19%), Pseudomonas aeruginosa (0.53%), Streptococcus pneumonia (0.4%)), Citrobacter (0.13%), Neisseria gonorrhea (0.13%), Enterococcus faecalis (0.13%), and Morganella morganii (0.13%). Infected patients were significantly more adults (73.1% vs. 21.5%) compared to children and mainly women (63.7% vs. 30.9%; p = 0.001). Conclusion: The observed expansion requires that hospital therapeutic committees set up an effective clinical management system and define the right combinations of antibiotics.

Keywords: multidrug resistance, bacteria, antibiogram, Bukavu

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Authors: Roxana Gheorghita, Gheorghe Gutt

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Environmental pollution due to non-degradation of packaging from the food and pharmaceutical industry is reaching increasingly alarming levels. The packaging used for food supplements is usually composed of successive layers of synthetic materials, conventional, glue, and paint. The situation is becoming more and more problematic as the population, according to statistics, uses food supplements more and more often. The solution can be represented by edible packaging, completely biodegradable, and compostable. The tested materials were obtained from biopolymers, agar, carrageenan, and alginate, in well-established quantities and plasticized with glycerol. Rosemary, thyme, and oregano essential oils have been added in varying proportions. The obtained films are completely water-soluble in hot liquids (with a temperature of about 80° C) and can be consumed with the product contained. The films were glossy, pleasant to the touch, thin (thicknesses between 32.8 and 52.8 μm), transparent, and with a pleasant smell, specific to the added essential oil. Tested for microbial evaluation, none of the films indicated the presence of E. coli, S. aureus, enterobacteria, coliform bacteria, yeasts, or molds. This aspect can also be helped by the low values of the water activity index (located between 0.546 and 0.576). The mechanical properties indicated that the material became more resistant with the addition of essential oil, the best values being recorded by the addition of oregano. The results obtained indicate the possibility of using biopolymer-based films with the addition of rosemary, thyme, and oregano essential oil, for wrapping food supplements, thus replacing conventional packaging, multilayer, impossible to sort and recycle.

Keywords: edible films, food supplements, oregano, rosemary, thyme

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Authors: Savitha Janakiraman, Shivakumar M. C

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Anidulafungin, an advanced class of antifungal agent used for the treatment of chronic fungal infections, is derived from Echinocandin B nucleus, an intermediate metabolite of Echinocandin B produced by Aspergillus nidulans. The enzyme acylase derived from the fermentation broth of Actinoplanes utahensis (NRRL 12052) plays a key role in the bioconversion of echinocandin B to echinocandin B nucleus. The membrane-bound nature of acylase and low levels of expression contributes to the rate-limiting process of enzymatic deacylation, hence low yields of ECB nucleus and anidulafungin. In the present study, this is addressed through novel genetic engineering approaches of overexpression and heterologous expression studies, immobilization of whole cells of Actinoplanes utahensis (NRRL 12052) and Co-cultivation studies. Overexpression of the acylase gene in Actinoplanes utahensis (NRRL 12052) was done by increasing the gene copy number to increase the echinocandin B nucleus production. Echinocandin B acylase gene, under the control of a PermE* promoter, was cloned in pSET152 vector and introduced into Actinoplanes utahensis (NRRL12052) by a ɸC31-directed site-specific recombination method. The resultant recombinant strain (C2-18) showed a 3-fold increase in acylase expression, which was confirmed by HPLC analysis. Pichia pastoris is one of the most effective and versatile host systems for the production of heterologous proteins. The ECB acylase gene was cloned into pPIC9K vector with AOX1 promoter and was transformed into Pichia pastoris (GS115). The acylase expression was confirmed by protein expression and bioconversion studies. The heterologous expression of acylase in Pichia pastoris, is a milestone in the development of antifungals. Actively growing cells of Actinoplanes utahensis (NRRL 12052) were immobilized and tested for bioconversion ability which showed >90% conversion in each cycle. The stability of immobilized cell beads retained the deacylation ability up to 60 days and reusability was confirmed up to 4 cycles. The significant findings from the study have revealed that immobilization of whole cells of Actinoplanes utahensis (NRRL 12052) could be an alternative option for bioconversion of echinocandin B to echinocandin B nucleus, which has not been reported to date. The concept of co-cultivation of Aspergillus nidulans and Actinoplanes utahensis strains for the production of the echinocandin B nucleus was also carried out in order to produce echinocandin B nucleus. The process completely reduced the ECB purification step and, therefore, could be recommended as an ingenious method to improve the yield of the ECB nucleus.

Keywords: acylase, anidulafungin, antifungals, Aspergillus nidulans

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Authors: Zainab Abbas Soharwardi, Chunli Su, Fazeelat Tahira, Syed Zahid Aziz

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The study monitors the quality of drinking water consumed by urban population of Lahore. A total of 50 drinking water samples (16 from source and 34 from distribution system) were examined for physical, chemical and bacteriological parameters. The parameters including pH, turbidity, electrical conductivity, total dissolved solids, total hardness, calcium, magnesium, total alkalinity, carbonate, sulphate, chloride, nitrite, fluoride, sodium and potassium were analyzed. Sixteen out of fifty samples showed high values of alkalinity compared to EPA standards and WHO guidelines. Twenty-eight samples were analyzed for heavy metals, chromium, iron, copper, zinc, cadmium and lead. Trace amounts of heavy metals were detected in some samples, however for most of the samples values were within the permissible limits although high concentration of zinc was detected in one sample collected from Mughal Pura area. Fifteen samples were analyzed for arsenic. The results were unsatisfactory; around 73% samples showed exceeding values of As. WHO has suggested permissible limits of arsenic < 0.01 ppm, whereas 27 % of samples have shown 0.05 ppm arsenic, which is five times greater than WHO highest permissible limits. All the samples were examined for E. coli bacteria. On the basis of bacteriological analysis, 42 % samples did not meet WHO guidelines and were unsafe for drinking.

Keywords: arsenic, heavy metals, ground water, Lahore

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Authors: Indrajeet Kumar, Jayanta Bhattacharya

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This study was evaluated for the potential use of nanosilver (nAg) as a disinfectant and antimicrobial growth promoter supplement for the poultry. The experiments were conducted in the Kangsabati river basin region, in West Medinipur district, West Bengal, India for six months. Two poultry farms were adopted for the experiment. The rural economy of this region from Jhargram to Barkola is heavily dependent on contract poultry farming. The water samples were collected from the water source of poultry farm which has been used for poultry drinking purpose. The bacteriological analysis of water sample revealed that the total bacterial count (total coliform and E. coli) were higher than the acceptable standards. The bacterial loads badly affected the growth performance and health of the poultry. For disinfection, a number of chemical compounds (like formaldehyde, calcium hypochloride, sodium hypochloride, and sodium bicarbonate) have been used in typical commercial formulations. However, the effects of all these chemical compounds have not been significant over time. As a part of our research-to-market initiative, we used nanosilver (nAg) formulation as a disinfectant. The nAg formulation was synthesized by hydrothermal technique and characterized by UV-visible, TEM, SEM, and EDX. The obtained results revealed that the mortality rate of poultry was reduced due to nAg formulation compared to the mortality rate of the negative control. Moreover, the income of the farmer family was increased by 10-20% due to less mortality and better health of the poultry.

Keywords: farm water, nanosilver, field application, and poultry performance

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Authors: Kirstin Burger, Paul Watts, Nicole Vorster

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Green chemistry focuses on synthesis which has a low negative impact on the environment. This research focuses on synthesizing novel compounds from an all-natural Eucalyptus citriodora oil. Eight novel plasticizer compounds are synthesized and optimized using flow chemistry technology. A precursor to one novel compound can be synthesized from the lauric acid present in coconut oil. Key parameters, such as catalyst screening and loading, reaction time, temperature, residence time using flow chemistry techniques is investigated. The compounds are characterised using GC-MS, FT-IR, 1H and 13C-NMR techniques, X-ray crystallography. The efficiency of the compounds is compared to two commercial plasticizers, i.e. Dibutyl phthalate and Eastman 168. Several PVC-plasticized film formulations are produced using the bio-based novel compounds. Tensile strength, stress at fracture and percentage elongation are tested. The property of having increasing plasticizer percentage in the film formulations is investigated, ranging from 3, 6, 9 and 12%. The diastereoisomers of each compound are separated and formulated into PVC films, and differences in tensile strength are measured. Leaching tests, flexibility, and change in glass transition temperatures for PVC-plasticized films is recorded. Research objective includes using these novel compounds as a green bio-plasticizer alternative in plastic products for infants. The inhibitory effect of the compounds on six pathogens effecting infants are studied, namely; Escherichia coli, Staphylococcus aureus, Shigella sonnei, Pseudomonas putida, Salmonella choleraesuis and Klebsiella oxytoca.

Keywords: bio-based compounds, plasticizer, tensile strength, microbiological inhibition , synthesis

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Authors: Michal Pravenec, Miroslava Simakova, Jan Silhavy

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Brown adipose tissue (BAT) plays an important role in lipid and glucose metabolism in rodents and possibly also in humans. Recently, using systems genetics approach in the BAT from BXH/HXB recombinant inbred strains, derived from the SHR (spontaneously hypertensive rat) and BN (Brown Norway) progenitors, we identified Cd36 (fatty acid translocase) as the hub gene of co-expression module associated with BAT relative weight and function. An important aspect of BAT biology is to better understand the mechanisms regulating the uptake and utilization of fatty acids and glucose. Accordingly, BAT function in the SHR that harbors mutant nonfunctional Cd36 variant (hereafter referred to as SHR-Cd36⁻/⁻) was compared with SHR transgenic line expressing wild type Cd36 under control of a universal promoter (hereafter referred to as SHR-Cd36⁺/⁺). BAT was incubated in media containing insulin and 14C-U-glucose alone or 14C-U-glucose together with palmitate. Incorporation of glucose into BAT lipids was significantly higher in SHR-Cd36⁺/⁺ versus SHR-Cd36⁻/⁻ rats when incubation media contained glucose alone (SHR-Cd36⁻/⁻ 591 ± 75 vs. SHR-Cd36⁺/⁺ 1036 ± 135 nmol/gl./2h; P < 0.005). Adding palmitate into incubation media had no effect in SHR-Cd36⁻/⁻ rats but significantly reduced glucose incorporation into BAT lipids in SHR-Cd36⁺/⁺ (SHR-Cd36⁻/⁻ 543 ± 55 vs. SHR-Cd36⁺/⁺ 766 ± 75 nmol/gl./2h; P < 0.05 denotes significant Cd36 x palmitate interaction determined by two-way ANOVA). This Cd36-dependent reduced glucose uptake in SHR-Cd36⁺/⁺ BAT was likely secondary to increased palmitate incorporation and utilization due to the presence of wild type Cd36 fatty acid translocase in transgenic rats. This possibility is supported by increased incorporation of 14C-U-palmitate into BAT lipids in the presence of both palmitate and glucose in incubation media (palmitate alone: SHR-Cd36⁻/⁻ 870 ± 21 vs. SHR-Cd36⁺/⁺ 899 ± 42; glucose+palmitate: SHR-Cd36⁻/⁻ 899 ± 47 vs. SHR-Cd36⁺/⁺ 1460 ± 111 nmol/palm./2h; P < 0.05 denotes significant Cd36 x glucose interaction determined by two-way ANOVA). It is possible that addition of glucose into the incubation media increased palmitate incorporation into BAT lipids in SHR-Cd36⁺/⁺ rats because of glucose availability for glycerol phosphate production and increased triglyceride synthesis. These changes in glucose and palmitate incorporation into BAT lipids were associated with significant differential expression of Irs1, Irs2, Slc2a4 and Foxo1 genes involved in insulin signaling and glucose metabolism only in SHR-Cd36⁺/⁺ rats which suggests Cd36-dependent effects on insulin action. In conclusion, these results provide compelling evidence that Cd36 plays an important role in BAT insulin signaling and energy substrate utilization.

Keywords: brown adipose tissue, Cd36, energy substrate utilization, insulin signaling, spontaneously hypertensive rat

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Authors: Yogesh Karunakar, Praveen Kandaswamy

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Mother’s milk provides the most superior source of nutrition to a child. There is no other substitute to the mother’s milk. Postpartum secretions like breast milk can be analyzed on the go for testing the presence of any harmful pathogen before a mother can feed the child or donate the milk for the milk bank. Since breast feeding is one of the main causes for transmission of diseases to the newborn, it is mandatory to test the secretions. In this paper, we describe the detection of pathogens like E-coli, Human Immunodeficiency Virus (HIV), Hepatitis B (HBV), Hepatitis C (HCV), Cytomegalovirus (CMV), Zika and Ebola virus through an innovative method, in which we are developing a unique chip for testing the mother’s milk sample. The chip will contain an antibody specific to the target pathogen that will show a color change if there are enough pathogens present in the fluid that will be considered dangerous. A smart-phone camera will then be acquiring the image of the strip and using various image processing techniques we will detect the color development due to antigen antibody interaction within 5 minutes, thereby not adding to any delay, before the newborn is fed or prior to the collection of the milk for the milk bank. If the target pathogen comes positive through this method, then the health care provider can provide adequate treatment to bring down the number of pathogens. This will reduce the postpartum related mortality and morbidity which arises due to feeding infectious breast milk to own child.

Keywords: postpartum, fluids, camera, HIV, HCV, CMV, Zika, Ebola, smart-phones, breast milk, pathogens, image processing techniques

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Authors: Sujata Sharma, Avinash Singh, Lovely Gautam, Pradeep Sharma, Mau Sinha, Asha Bhushan, Punit Kaur, Tej P. Singh

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Peptidyl-tRNA hydrolase (Pth) Pth is an essential bacterial enzyme that catalyzes the release of free tRNA and peptide moeities from peptidyl tRNAs during stalling of protein synthesis. In order to design inhibitors of Pth from Pseudomonas aeruginosa (PaPth), we have determined the structures of PaPth in its native state and in the bound states with two compounds, amino acylate-tRNA analogue (AAtA) and 5-azacytidine (AZAC). The peptidyl-tRNA hydrolase gene from Pseudomonas aeruginosa was amplified by Phusion High-Fidelity DNA Polymerase using forward and reverse primers, respectively. The E. coliBL21 (λDE3) strain was used for expression of the recombinant peptidyl-tRNA hydrolase from Pseudomonas aeruginosa. The protein was purified using a Ni-NTA superflow column. The crystallization experiments were carried out using hanging drop vapour diffusion method. The crystals diffracted to 1.50 Å resolution. The data were processed using HKL-2000. The polypeptide chain of PaPth consists of 194 amino acid residues from Met1 to Ala194. The centrally located β-structure is surrounded by α-helices from all sides except the side that has entrance to the substrate binding site. The structures of the complexes of PaPth with AAtA and AZAC showed the ligands bound to PaPth in the substrate binding cleft and interacted with protein atoms extensively. The residues that formed intermolecular hydrogen bonds with the atoms of AAtA included Asn12, His22, Asn70, Gly113, Asn116, Ser148, and Glu161 of the symmetry related molecule. The amino acids that were involved in hydrogen bonded interactions in case of AZAC included, His22, Gly113, Asn116, and Ser148. As indicated by fittings of two ligands and the number of interactions made by them with protein atoms, AAtA appears to be a more compatible with the structure of the substrate binding cleft. However, there is a further scope to achieve a better stacking than that of O-tyrosyl moiety because it is not still ideally stacked. These observations about the interactions between the protein and ligands have provided the information about the mode of binding of ligands, nature and number of interactions. This information may be useful for the design of tight inhibitors of Pth enzymes.

Keywords: peptidyl tRNA hydrolase, Acinetobacter baumannii, Pth enzymes, O-tyrosyl

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Authors: Merih Şimşek, Recep Keşli, Özgül Çetinkaya, Cengiz Demir, Adem Aslan

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Bacteremia is a clinical entity with high morbidity and mortality rates when immediate diagnose, or treatment cannot be achieved. Microorganisms which can cause sepsis or bacteremia are easily isolated from blood cultures. Fifty-five positive blood cultures were included in this study. Microorganisms in 55 blood cultures were isolated by conventional microbiological methods; afterwards, microorganisms were defined in terms of the phenotypic aspects by the Vitek-2 system. The same microorganisms in all blood culture samples were defined in terms of genotypic aspects again by Multiplex-PCR DNA Low-Density Microarray System. At the end of the identification process, the DNA microarray system’s success in identification was evaluated based on the Vitek-2 system. The Vitek-2 system and DNA Microarray system were able to identify the same microorganisms in 53 samples; on the other hand, different microorganisms were identified in the 2 blood cultures by DNA Microarray system. The microorganisms identified by Vitek-2 system were found to be identical to 96.4 % of microorganisms identified by DNA Microarrays system. In addition to bacteria identified by Vitek-2, the presence of a second bacterium has been detected in 5 blood cultures by the DNA Microarray system. It was identified 18 of 55 positive blood culture as E.coli strains with both Vitek 2 and DNA microarray systems. The same identification numbers were found 6 and 8 for Acinetobacter baumanii, 10 and 10 for K.pneumoniae, 5 and 5 for S.aureus, 7 and 11 for Enterococcus spp, 5 and 5 for P.aeruginosa, 2 and 2 for C.albicans respectively. According to these results, DNA Microarray system requires both a technical device and experienced staff support; besides, it requires more expensive kits than Vitek-2. However, this method should be used in conjunction with conventional microbiological methods. Thus, large microbiology laboratories will produce faster, more sensitive and more successful results in the identification of cultured microorganisms.

Keywords: microarray, Vitek-2, blood culture, bacteremia

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Authors: Sachin Latiyan, T. S. Sampath Kumar, Mukesh Doble

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Natural polymer based nanofibrous wound dressings have gained increased attention because of their high surface area, bioactivity, biodegradability and resemblance to extracellular matrix. Agarose (a natural polymer) have been used largely for angiogenesis, cartilage formation and wound healing applications. However, electrospinning of agarose is tedious thereby rendering limited studies on fabrication and evaluation of agarose based nanofibrous wound dressings. Thus, present study focuses on the fabrication of agarose (10% w/v)/ polyvinyl alcohol (12% w/v) based multifunctional nanofibrous scaffolds. Zinc citrate (1, 3 and 5% w/w of the polymer) was added as a potential antibacterial agent to combat wound infections. The fabricated scaffolds exhibit ~500% swelling (in phosphate buffer saline) with enhanced mechanical strength which is suitable for most of the wound healing applications. In vitro studies were found to reveal an increased migration and proliferation of L929 mouse fibroblasts with agarose blends w.r.t to the control. The fabricated dressings were found to be effective against both Escherichia coli (Gram-negative) and Staphylococcus aureus (Gram-positive) bacterial strains. Hence, a multifunctional (as provides effective swelling and mechanical support along with antibacterial property), natural product based, eco-friendly scaffold was successfully fabricated to serve as a potential wound dressing material.

Keywords: antibacterial dressings, benign solvent, nanofibrous agarose, biocompatibility, enhanced swelling and mechanical strength, biopolymeric dressings

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Authors: Anupalli Roja Rani, Saraswathi Jaggali

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Rose-scented geranium (Pelargonium graveolens L’Hér.) is an erect, much-branched shrub. It is indigenous to various parts of southern Africa, and it is often called Geranium. Pelargonium species are widely used by traditional healers in the areas of Southern Africa by Sotho, Xhosa, Khoi-San and Zulus for its curative and palliative effects in the treatment of diarrhea, dysentery, fever, respiratory tract infections, liver complaints, wounds, gastroenteritis, haemorrhage, kidney and bladder disorders. We have used Plant materials for extracting active compounds from analytical grades of solvents methanol, ethyl acetate, chloroform and water by a soxhlet apparatus. The phytochemical screening reveals that extracts of Pelargonium graveolens contains alkaloids, glycosides, steroids, tannins, saponins and phenols in ethyl acetate solvent. The antioxidant activity was determined using 1, 1-diphenyl-2-picrylhydrazyl (DPPH) bleaching method and the total phenolic content in the extracts was determined by the Folin–Ciocalteu method. Due to the presence of different phytochemical compounds in Pelargonium the anti-microbial activity against different micro-organisms like E.coli, Streptococcus, Klebsiella and Bacillus. Fractionation of plant extract was performed by column chromatography and was confirmed with HPLC analysis, NMR and FTIR spectroscopy for the compound identification in different organic solvent extracts.

Keywords: Pelargonium graveolens L’Hér, DPPH, micro-organisms, HPLC analysis, NMR, FTIR spectroscopy

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Authors: Saba Atefyekta

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Aim: Control of bacterial bioburden in wounds is an important step for minimizing the risk of wound infection. An antimicrobial hydrogel wound dressing is developed out of soft polymeric hydrogels that contain antimicrobial peptides (AMPs). Such wound dressings can bind and kill all types of bacteria, even the resistance types at the wound site. Methods: AMPs are permanently bonded onto a soft nanostructured polymer via covalent attachment and physical entanglement. This improves stability, rapid antibacterial activity, and, most importantly, prevents the leaching of AMPs. Major Findings: Antimicrobial analysis of antimicrobial hydrogels using in-vitro wound models confirmed >99% killing efficiency against multiple bacterial trains, including MRSA, MDR, E. Coli. Furthermore, the hydrogel retained its antibacterial activity for up to 4 days when exposed to human serum. Tests confirmed no release of AMPs, and it was proven non-toxic to mammalian cells. An in-vivo study on human intact skin showed a significant reduction of bacteria for part of the subject’s skin treated with antibacterial hydrogels. A similar result was detected through a qualitative study in veterinary trials on different types of surgery wounds in cats, dogs, and horses. Conclusions: Antimicrobial hydrogels wound dressings developed by permanent attachment of AMPs can effectively and rapidly kill bacteria in contact. Such antibacterial hydrogel wound dressings are non-toxic and do not release any substances into the wound.

Keywords: antibacterial wound dressing, antimicrobial peptides, post-surgical wounds, infection

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Authors: Sarita Puri, Tapan K. Chaudhuri

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Chaperonin GroEL is a tetradecameric Escherichia coli protein having identical subunits of 57 kDa. The elucidation of thermodynamic parameters related to stability for the native GroEL is not feasible as it undergoes irreversible unfolding because of its large size (800kDa) and multimeric nature. Nevertheless, it is important to determine the thermodynamic stability parameters for the highly stable GroEL protein as it helps in folding and holding of many substrate proteins during many cellular stresses. Properly folded monomers work as building-block for the formation of native tetradecameric GroEL. Spontaneous refolding behavior of monomeric GroEL makes it suitable for protein-denaturant interactions and thermodynamic stability based studies. The urea mediated unfolding is a three state process which means there is the formation of one intermediate state along with native and unfolded states. The heat mediated denaturation is a two-state process. The unfolding process is reversible as observed by the spontaneous refolding of denatured protein in both urea and head mediated refolding processes. Analysis of folding/unfolding data provides a measure of various thermodynamic stability parameters for the monomeric GroEL. The proposed mechanism of unfolding of monomeric GroEL is a three state process which involves formation of one stable intermediate having folded apical domain and unfolded equatorial, intermediate domains. Research in progress is to demonstrate the importance of specific residues in stability and oligomerization of GroEL protein. Several mutant versions of GroEL are under investigation to resolve the above mentioned issue.

Keywords: equilibrium unfolding, monomeric GroEl, spontaneous refolding, thermodynamic stability

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Authors: E. Bartkiene, M. Ruzauskas, V. Lele, P. Zavistanaviciute, J. Bernatoniene, V. Jakstas, L. Ivanauskas, D. Zadeike, D. Klupsaite, P. Viskelis, J. Bendoraitiene, V. Navikaite-Snipaitiene, G. Juodeikiene

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In this study, antimicrobial nutraceuticals; gummy candies (GC) from bovine colostrum (BC), essential oils (EOs), probiotic lactic acid bacteria (PLAB), and their combinations, were developed. For antimicrobial GC preparation, heteropolysaccharide (agar) was used. The antimicrobial properties of EOs (Eugenia caryophyllata, Thymus vulgaris, Citrus reticulata L., Citrus paradisi L.), BC, L. paracasei LUHS244, L. plantarum LUHS135, and their combinations against pathogenic bacteria strains (Streptococcus mutans, Enterococcus faecalis, Staphylococcus aureus, Salmonella enterica, Escherichia coli, Proteus mirabilis, and Pseudomonas aeruginosa) were evaluated. The highest antimicrobial properties by EO’s (Eugenia caryophyllata and Thymus vulgaris) were established. The optimal ingredients composition for antimicrobial GC preparation was established, which incorporate the BC fermented with L. paracasei LUHS244 in combination with Thymus vulgaris or Eugenia caryophyllata. These ingredients showed high inhibition properties of all tested pathogenic strains (except Pseudomonas aeruginosa). Antimicrobial GC formula consisting of thyme EO (up to 0.2%) and fermented BC (up to 3%), and for taste masking, mandarin or grapefruit EOs (up to 0.2%) was used. Developed GC high overall acceptability and antimicrobial properties, thus, antimicrobial GC could be a preferred form of nutraceuticals. This study was fulfilled with the support of the LSMU-KTU joint project.

Keywords: antimicrobial activity, bovine colostrum, essential oil, gummy candy, probiotic

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