Search results for: recombinant protein purification

Authors: Debananda Mohapatra, Subramanya Badrayyana, Smrutiranjan Parida

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Recognizing the upcoming era of carbon nanostructures and their revolutionary applications, we investigated the formation and supercapacitor application of highly pure and hydrophilic carbon nano-onions (CNOs) by economical one-step flame-synthesis procedure. The facile and scalable method uses easily available organic carbon source such as clarified butter, restricting the use of any catalyst, sophisticated instrumentation, high vacuum and post processing purification procedure. The active material was conformally coated onto a locally available cotton wipe by “sonicating and drying” process to obtain novel, lightweight, inexpensive, flexible, binder-free electrodes with strong adhesion between nanoparticles and porous wipe. This interesting electrode with CNO as the active material delivers a specific capacitance of 102.16 F/g, the energy density of 14.18 Wh/kg and power density of 2448 W/kg which are the highest values reported so far in symmetrical two electrode cell configuration with 1M Na2SO4 as an electrolyte. Incorporation of CuO nanoparticles to these functionalized CNOs by one-step hydrothermal method add up to a significant specific capacitance of 420 F/g with deliverable energy and power density at 58.33 Wh/kg and 4228 W/kg, respectively. The free standing CNOs, as well as CNO-CuO composite electrode, showed an excellent cyclic performance and stability retaining 95 and 90% initial capacitance even after 5000 charge-discharge cycles at a current density of 5 A/g. This work presents a new platform for high performance supercapacitors for next generation wearable electronic devices.

Keywords: binder-free, flame synthesis, flexible, carbon nano-onion

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Authors: Mojdeh Chelemal Dezfoul Nejad, Masomeh Moradi, Mehrzad Mesbah, Mehran Javaheri Babouli

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This study was conducted in order to characterize the different levels of dietary vitamin C on some of biochemical parameters of Barbuas grypus. For this purpose 300 Barbuas grypus were divided into 15 groups. five levels of vitamin C (0, 200 ,400,800,1600 mg kg-1 diet) and their combination were used to prepare five experimental diets. The fish were fed 3% of their wet b.wt. per day for a 60 days period. Blood samples were obtained from six fish of each tank at the end of experiment. The results reveal that fish fed diets containing 1600 mg kg^-1 vitamin C had the significant decreased in the mean amount of cholesterol, glucose and triglyceride (p<0.05). Also, there was no significant difference in the mean amount of total protein with the different diets designed for this experiment (p>0.05).

Keywords: Barbuas, grypus, vitamin C, biochemical parameters

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Authors: Tarnveer Kaur

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Amino acids, as fundamental structural units of peptides and proteins, have an important role in biological systems by affecting solubility, denaturation, and activity of biomolecules. A study of these effects on thermophysical properties of model compounds in the presence of electrolytes solutions provides information about solute-solvent and solute-solute interactions on biomolecules. Ionic liquids (ILs) as organic electrolytes and green solvents are composed of an organic cation and an inorganic anion, which are liquid at ambient conditions. In the past decade, extensive investigations showed that the use of ILs as reaction media for processes involving biologically relevant compounds is promising in view of their successful application in kinetic resolution, biocatalysis, biosynthesis, separation, and purification processes. The scope of this information is valuable to explore the interactions of amino acids in ILs. To reach this purpose, apparent molar volumes of glycine/L-alanine in aqueous solutions of 1-butyl-3-methylimidazolium bromide/chloride were determined from precise density measurements at temperatures T = (288.15-318.15) K and at atmospheric pressure. Positive values for all the studied amino acids indicate the dominance of hydrophilic-ionic interactions between amino acids and Ionic liquids. The effect of temperature on volumetric properties of glycine/L-alanine in solutions has been determined from the partial molar expansibility and second-order partial molar expansibility. Further, volumetric interaction parameters and hydration number have been calculated, which have been interpreted in terms of possible solute-solvent interactions.

Keywords: ILs, amino acids, volumetric properties, hydration numbers

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Authors: Deepanjali Gurav, Kun Qian

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In-vitro metabolic diagnosis relies on designed materials-based analytical platforms for detection of selected metabolites in biological samples, which has a key role in disease detection and therapeutic evaluation in clinics. However, the basic challenge deals with developing a simple approach for metabolic analysis in bio-samples with high sample complexity and low molecular abundance. In this work, we report a designer plasmonic nanoshells based platform for direct detection of small metabolites in clinical samples for in-vitro metabolic diagnostics. We first synthesized a series of plasmonic core-shell particles with tunable nanoshell structures. The optimized plasmonic nanoshells as new matrices allowed fast, multiplex, sensitive, and selective LDI MS (Laser desorption/ionization mass spectrometry) detection of small metabolites in 0.5 μL of bio-fluids without enrichment or purification. Furthermore, coupling with isotopic quantification of selected metabolites, we demonstrated the use of these plasmonic nanoshells for disease detection and therapeutic evaluation in clinics. For disease detection, we identified patients with postoperative brain infection through glucose quantitation and daily monitoring by cerebrospinal fluid (CSF) analysis. For therapeutic evaluation, we investigated drug distribution in blood and CSF systems and validated the function and permeability of blood-brain/CSF-barriers, during therapeutic treatment of patients with cerebral edema for pharmacokinetic study. Our work sheds light on the design of materials for high-performance metabolic analysis and precision diagnostics in real cases.

Keywords: plasmonic nanoparticles, metabolites, fingerprinting, mass spectrometry, in-vitro diagnostics

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Authors: Somayeh Fani, Mahmood Ameen Abdulla

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Costus speciosus is an important Malaysian medicinal plant commonly used traditionally in the treatment of many aliments. The present investigation is designed to elucidate preventive effects of ethanolic extracts of C. speciosus rhizome against absolute ethanol-induced gastric mucosal injury in Sprague-Dawley rats. Five groups of rats were orally pre-treated with vehicle, carboxymethylcellulose (CMC) as normal control group (Group 1), ethanol as ulcer control group (Group 2), omeprazole 20 mg/kg (reference group) (Group 3), and 250 and 500 mg/kg of C. speciosus extract (experimental groups) (Group 4 and 5), respectively. An hour later, CMC was given orally to Group 1 rats and absolute ethanol was given orally to Group 2-5 rats to generate gastric mucosal injury. After an additional hour, the rats were sacrificed. Grossly, ulcer control group exhibited severe of gastric mucosal hemorrhagic injury and increased in ulcer area, whereas groups pre-treated with omeprazole or plant’s rhizomes exhibited the significant reduction of gastric mucosal injury. Significant increase in the pH and mucous of gastric content was observed in rats re-treated with C. speciosus rhizome. Histology, ulcer control rats, demonstrated remarkable disruption of gastric mucosa, increased in edema and inflammatory cells infiltration of submucosal layer compared to rats pre-treated with rhizomes extract. Periodic acid Schiff staining for glycoprotein, rats pre-fed with C. speciosus C. displayed remarkably intense uptake of magenta color by glandular gastric mucosa compared with ulcer control rats. Immunostaining of gastric epithelium, rats pre-treatment with rhizome extract provide evidence of up-regulation of HSP70 and down-regulation of Bax proteins compared to ulcer control animals. Gastric tissue homogenate, C. speciosus significantly increased the activity of superoxide dismutase (SOD), and catalase (CAT), increased the level of non-protein sulfhydryl (NP-SH) and decreased the level of lipid peroxidation after ethanol administration. Acute toxicity test did not show any signs of toxicity. The mechanisms implicated the gasrtoprotective property of C. speciosus depend upon the antisecretory activity, increased in gastric mucus glycoprotein, up-regulation of HSP70 protein and down-regulation of Bax proteins, reduction in the lipid peroxidation and increase in the level of NP-SH and antioxidant enzymes activity in gastic homogenate.

Keywords: antioxidant, Costus speciosus, gastric ulcer, histology, omeprazole

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Authors: A. Ferasat, S. Rostampour Yasouri

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Seasonal flu is a highly contagious infection caused by influenza viruses. These viruses undergo genetic changes that result in new epidemics across the globe. Medical attention is crucial in severe cases, particularly for the elderly, frail, and those with chronic illnesses, as their immune systems are often weaker. The purpose of this study was to detect new subtypes of the influenza A virus rapidly using a specific RT-PCR method based on the HA gene (hemagglutinin). In the winter and spring of 2022_2023, 120 embryonated egg samples were cultured, suspected of seasonal influenza. RNA synthesis, followed by cDNA synthesis, was performed. Finally, the PCR technique was applied using a pair of specific primers designed based on the HA gene. The PCR product was identified after purification, and the nucleotide sequence of purified PCR products was compared with the sequences in the gene bank. The results showed a high similarity between the sequence of the positive samples isolated from the patients and the sequence of the new strains isolated in recent years. This RT-PCR technique is entirely specific in this study, enabling the detection and multiplication of influenza and its subspecies from clinical samples. The RT-PCR technique based on the HA gene, along with sequencing, is a fast, specific, and sensitive diagnostic method for those infected with influenza viruses and its new subtypes. Rapid molecular diagnosis of influenza is essential for suspected people to control and prevent the spread of the disease to others. It also prevents the occurrence of secondary (sometimes fatal) pneumonia that results from influenza and pathogenic bacteria. The critical role of rapid diagnosis of new strains of influenza is to prepare a drug vaccine against the latest viruses that did not exist in the community last year and are entirely new viruses.

Keywords: influenza, molecular diagnosis, patients, RT-PCR technique

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Authors: Shu-Jen Chen, Yi-Chien Wan, Ruey-Chi Wang

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Hydroxyapatite (Ca10(PO4)6(OH)2, HAp) can be applied to the fabrication of bone replacement materials, the composite of dental filling, and the adsorption of biomolecules and dyes. The integration of HAp and magnetic materials would offer several advantages for bio-separation process because the magnetic adsorbents is capable of recovered by applied magnetic field. C-phycocyanin (C-PC) and Allophycocyanin (APC), isolated from Spirulina platensis, can be used in fluorescent labeling probes, health care foods and clinical diagnostic reagents. Although the purification of C-PC and APC are reported by HAp adsorption, the adsorption of C-PC and APC by magnetic HAp composites was not reported yet. Therefore, the fabrication of HAp with magnetic silica nanoparticles for proteins adsorption was investigated in this work. First, the magnetic silica particles were prepared by covering silica layer on Fe3O4 nanoparticles with a reverse micelle method. Then, the Fe3O4@SiO2 nanoparticles were mixed with calcium carbonate to obtain magnetic silica/calcium carbonate composites (Fe3O4@SiO2/CaCO3). The Fe3O4@SiO2/CaCO3 was further reacted with K2HPO4 for preparing the magnetic silica/hydroxyapatite composites (Fe3O4@SiO2/HAp). The adsorption experiments indicated that the adsorption capacity of Fe3O4@SiO2/HAp toward C-PC and APC were highest at pH 6. The adsorption of C-PC and APC by Fe3O4@SiO2/HAp could be correlated by the pseudo-second-order model, indicating chemical adsorption dominating the adsorption process. Furthermore, the adsorption data showed that the adsorption of Fe3O4@SiO2/HAp toward C-PC and APC followed the Langmuir isotherm. The isoelectric points of C-PC and APC were around 5.0. Additionally, the zeta potential data showed the Fe3O4@SiO2/HAp composite was negative charged at pH 6. Accordingly, the adsorption mechanism of Fe3O4@SiO2/HAp toward C-PC and APC should be governed by hydrogen bonding rather than electrostatic interaction. On the other hand, as compared to C-PC, the Fe3O4@SiO2/HAp shows higher adsorption affinity toward APC. Although the Fe3O4@SiO2/HAp cannot recover C-PC and APC from Spirulina platensis homogenate, the Fe3O4@SiO2/HAp can be applied to separate C-PC and APC.

Keywords: hydroxyapatite, magnetic, C-phycocyanin, allophycocyanin

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Authors: Luz Stella Cadavid-Rodriguez, Viviana E. Castro-Lopez

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Fish waste (FW) has a high content of potentially biodegradable components, so it is amenable to be digested anaerobically. In this line, anaerobic digestion (AD) of FW has been studied for biogas production. Nevertheless, intermediate products such as volatile fatty acids (VFA), generated during the acidogenic stage, have been scarce investigated, even though they have a high potential as a renewable source of carbon. In the literature, there are few studies about the Inoculum-Substrate (I/S) ratio on acidogenesis. On the other hand, it is well known that pH is a critical factor in the production of VFA. The optimum pH for the production of VFA seems to change depending on the substrate and can vary in a range between 5.25 and 11. Nonetheless, the literature about VFA production from protein-rich waste, such as FW, is scarce. In this context, it is necessary to deepen on the determination of the optimal operating conditions of acidogenic fermentation for VFA production from protein-rich waste. Therefore, the aim of this research was to optimize the volatile fatty acid production from artisanal fishing waste, studying the effect of pH and the I/S ratio on the acidogenic process. For this research, the inoculum used was a methanogenic sludge (MS) obtained from a UASB reactor treating wastewater of a slaughterhouse plant, and the FW was collected in the port of Tumaco (Colombia) from the local artisanal fishers. The acidogenic fermentation experiments were conducted in batch mode, in 500 mL glass bottles as anaerobic reactors, equipped with rubber stoppers provided with a valve to release biogas. The effective volume used was 300 mL. The experiments were carried out for 15 days at a mesophilic temperature of 37± 2 °C and constant agitation of 200 rpm. The effect of 3 pH levels: 5, 7, 9, coupled with five I/S ratios, corresponding to 0.20, 0.15, 0.10, 0.05, 0.00 was evaluated taking as a response variable the production of VFA. A complete randomized block design was selected for the experiments in a 5x3 factorial arrangement, with two repetitions per treatment. At the beginning and during the process, pH in the experimental reactors was adjusted to the corresponding values of 5, 7, and 9 using 1M NaOH or 1M H2SO4, as was appropriated. In addition, once the optimum I/S ratio was determined, the process was evaluated at this condition without pH control. The results indicated that pH is the main factor in the production of VFA, obtaining the highest concentration with neutral pH. By reducing the I/S ratio, as low as 0.05, it was possible to maximize VFA production. Thus, the optimum conditions found were natural pH (6.6-7.7) and I/S ratio of 0.05, with which it was possible to reach a maximum total VFA concentration of 70.3 g Ac/L, whose major components were acetic acid (35%) and butyric acid (32%). The findings showed that the acidogenic fermentation of FW is an efficient way of producing VFA and that the operating conditions can be simple and economical.

Keywords: acidogenesis, artisanal fishing waste, inoculum to substrate ratio, volatile fatty acids

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Authors: K. V. Ramanaiah, R. Jagan, N. N. Murthy

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Trinuclear oxo-centered carboxylate-bridged iron complexes, [Fe3(µ3-O)(µ2-O2CR)L¬3]+/0 (where R = alkyl or aryl; L = H2O, ROH, Py, solvent) have attracted tremendous attention because of their interesting structural and magnetic properties, exhibit mixed-valent trapped and de-trapped states, and have bioinorganic relevance. The presence of a trinuclear iron binding center has been implicated in the formation of both bacterial and human iron storage protein, Ft. They are used as precursors for the synthesis of models for the active-site structures of non-heme proteins, hemerythrin (Hr), methane monooxygenase (MMO) and polyiron storage protein, ferritin (Ft). Used as important building blocks for the design and synthesis of supramolecules this can exhibit single molecular magnetism (SMM). Such studies have often employed simple and compact carboxylate ligands and the use of bulky carboxylates is scarce. In the present study, we employed two different type of sterically hindered carboxylates and synthesized a series of novel oxo-centered, carboxylate-bridged triiron complexes of general formula [Fe3(O)(O2CCPh3)6L3]X (L = H2O, 1; py, 2; 4-NMe2py, 3; X = ClO4; L = CH3CN, 4; X = FeCl4) and [Fe3(O)(O2C-anth)6L3]X (L = H2O, 5; X = ClO4; L = CH3OH, 6; X = Cl). Along with complex [Fe(OMe)2(O2CCPh3)]10, 7 was prepared by the self-assemble of anhydrous FeCl3, sodium triphenylacetate and sodium methoxide at ratio of 1:1:2 in CH3OH. The Electronic absorption spectra of these complexes 1-6, in CH2Cl2 display weak bands at near FTIR region (970-1135 nm, ε > 15M-1cm-1). For complex 7, one broad band centered at ~670nm and also an additional intense charge transfer (L→M or O→M) bands between 300 to 550nm observed for all the complexes. Paramagnetic 1H NMR is introduced as a good probe for the characterization of trinuclear oxo - cantered iron compounds in solution when the L ligand coordinated to iron varies as: H2O, py, 4-NMe2py, and CH3OH. The solution state magnetic moment values calculated by using Evans method for all the complexes and also solid state magnetic moment value of complex, 7 was calculated by VSM method, which is comparable with solution state value. These all magnetic moment values indicate there is a spin exchange process through oxo and carboxylate bridges in between two irons (d5). The ESI-mass data complement the data obtained from single crystal X-ray structure. Further purity of the compounds was confirmed by elemental analysis. Finally, structural determination of complexes 1, 3, 4, 5, 6 and 7 were unambiguously conformed by single crystal x-ray studies.

Keywords: decanuclear, paramagnetic NMR, trinuclear, uv-visible

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Authors: Mukunda Upreti, Jill Storry, Rafael Björk, Emilie Louvet, Johan Normark, Sven Bergström

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Borrelia duttonii and most other relapsing fever species are Gram-negative bacteria which cause a blood borne infection characterized by the binding of bacterium to erythrocytes. The bacteria associate with two or more erythrocytes to form clusters of cells into rosettes. Rosetting is a major virulence factor and the mechanism is believed to facilitate persistence of bacteria in the circulatory system and the avoidance of host immune cells through masking or steric hindrance effects. However, the molecular mechanisms of rosette formation are still poorly understood. This study aims at determining the molecules involved in the rosette formation phenomenon. Fractionated serum, using different affinity purification methods, was investigated as a rosetting agent and IgG and at least one other serum components were needed for rosettes to form. An IgG titration curve demonstrated that IgG alone is not enough to restore rosette formation level to the level whole serum gives. IgG hydrolysis by IdeS ( Immunoglobulin G-degrading enzyme of Streptococcus pyogenes) and deglycosylation using N-Glycanase proved that the whole IgG molecule regardless of saccharide moieties is critical for Borrelia induced rosetting. Complement components C3 and C4 were also important serum molecules necessary to maintain optimum rosetting rates. The deactivation of complement network and serum depletion with C3 and C4 significantly reduced the rosette formation rate. The dependency of IgG and complement components also implied involvement of the complement receptor (CR1). Rosette formation test with Knops null RBC and sCR1 confirmed that CR1 is also part of Borrelia induced rosette formation.

Keywords: complement components C3 and C4, complement receptor 1, Immunoglobulin G, Knops null, Rosetting

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Authors: Claudia Cabezas-Zabala, Jairo Lindarte-Artunduaga, Carlos Mario Zuluaga-Dominguez

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Amaranth is an emerging pseudocereal rich in such essential nutrients as protein and dietary fiber, which was employed as an ingredient in the formulation of crackers to evaluate the rheological performance and sensory acceptability of the obtained food. A completely randomized factorial design was used with two factors: (A) ratio of wheat and amaranth flour used in the preparation of the dough, in proportion 90:10 and 80:20 (% w/w) and (B) two levels of inulin addition of 8.4% and 16.7 %, having two control doughs made from amaranth and wheat flour, respectively. Initially, the functional properties of the formulations mentioned were measured, showing no significant differences in the water absorption capacity (WAC) and swelling power (SP), having mean values between 1.66 and 1.81 g/g for WAC and between 1.75 and 1.86 g/g for SP, respectively. The amaranth flour had the highest water holding capacity (WHR) of 8.41 ± 0.15 g/g and emulsifying activity (EA) of 74.63 ± 1.89 g/g. Moreover, the rheological behavior, measured through the use of farinograph, extensograph, Mixolab, and falling index, showed that the formulation containing 20% of amaranth flour and 7.16% of inulin had a rheological behavior similar to the control produced exclusively with wheat flour, being the former, the one selected for the preparation of crackers. For this formulation, the farinograph showed a mixing tolerance index of 11 UB, indicating a strong and cohesive dough; likewise, the Mixolab showed dough reaches stability at 6.47 min, indicating a good resistance to mixing. On the other hand, the extensograph exhibited a dough resistance of 637 UB, as well as extensibility of 13.4 mm, which corresponds to a strong dough capable of resisting the laminate. Finally, the falling index was 318 s, which indicates the crumb will retain enough air to enhance the crispness of a characteristic cracker. Finally, a sensory consumer test did not show significant differences in the evaluation of aroma between the control and the selected formulation, while this latter had a significantly lower rating in flavor. However, a purchase intention of 70 % was observed among the population surveyed. The results obtained in this work give perspectives for the industrial use of amaranth in baked goods. Additionally, amaranth has been a product typically linked to indigenous populations in the Andean South American countries; therefore, the search for diversification and alternatives of use for this pseudocereal has an impact on the social and economic conditions of such communities. The technological versatility and nutritional quality of amaranth is an advantage for consumers, favoring the consumption of healthy products with important contributions of dietary fiber and protein.

Keywords: amaranth, crackers, rheology, pseudocereals, kneaded products

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Authors: Marcin Stasiulewicz, Sebastian Filipkowski, Aneta Panuszko

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Background: The hallmark of neurodegenerative diseases, including Alzheimer's and Parkinson's diseases, is an aggregation of the abnormal forms of peptides and proteins. Water is essential to functioning biomolecules, and it is one of the key factors influencing protein folding and misfolding. However, the hydration studies of proteins are complicated due to the complexity of protein systems. The use of model compounds can facilitate the interpretation of results involving larger systems. Objectives: The goal of the research was to characterize the properties of the hydration water surrounding the two three-residue K peptide fragments INS (Isoleucine - Asparagine - Serine) and NSR (Asparagine - Serine - Arginine). Methods: Fourier-transform infrared spectra of aqueous solutions of the tripeptides were recorded on Nicolet 8700 spectrometer (Thermo Electron Co.) Measurements were carried out at 25°C for varying molality of solute. To remove oscillation couplings from water spectra and, consequently, obtain narrow O-D semi-heavy water bands (HDO), the isotopic dilution method of HDO in H₂O was used. The difference spectra method allowed us to isolate the tripeptide-affected HDO spectrum. Results: The structural and energetic properties of water affected by the tripeptides were compared to the properties of pure water. The shift of the values of the gravity center of bands (related to the mean energy of water hydrogen bonds) towards lower values with respect to the ones corresponding to pure water suggests that the energy of hydrogen bonds between water molecules surrounding tripeptides is higher than in pure water. A comparison of the values of the mean oxygen-oxygen distances in water affected by tripeptides and pure water indicates that water-water hydrogen bonds are shorter in the presence of these tripeptides. The analysis of differences in oxygen-oxygen distance distributions between the tripeptide-affected water and pure water indicates that around the tripeptides, the contribution of water molecules with the mean energy of hydrogen bonds decreases, and simultaneously the contribution of strong hydrogen bonds increases. Conclusions: It was found that hydrogen bonds between water molecules in the hydration sphere of tripeptides are shorter and stronger than in pure water. It means that in the presence of the tested tripeptides, the structure of water is strengthened compared to pure water. Moreover, it has been shown that in the vicinity of the Asparagine - Serine - Arginine, water forms stronger and shorter hydrogen bonds. Acknowledgments: This work was funded by the National Science Centre, Poland (grant 2017/26/D/NZ1/00497).

Keywords: amyloids, K-peptide, hydration, FTIR spectroscopy

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Authors: Ž. Vaštag, Lj. Popović, S. Popović

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After cold pressing of pumpkin oil, the defatted oil cake (PUOC) was utilized as raw material for processing of bio-functional hydrolysates. In this study, the in vitro bioactivity of an alcalase (AH) and a pepsin hydrolysate (PH) prepared from the major pumpkin 12S globulin (cucurbitin) are compared. The hydrolysates were produced at optimum reaction conditions (temperature, pH) for the enzymes, during 60min. The bioactivity testing included antioxidant and angiotensin I converting enzyme inhibitory activity assays. The hydrolysates showed high potential as natural antioxidants and possibly antihypertensive agents in functional food or nutraceuticals. Additionally, preliminary studies have shown that both hydrolysates could exhibit modest α-amylase inhibitory activity, which indicates on their hypoglycemic potential.

Keywords: cucurbitin, alcalase, pepsin, protein hydrolysates, in vitro bioactivity

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Authors: Asma Bashir, Neha Farid, Kashif Ali, Kiran Fatima

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Background: Bacteriocins are a subclass of antimicrobial peptides that are becoming extremely important in treatments. It is possible to utilise bacteriocins in place of or in addition to traditional antibiotics. It is possible to treat a variety of infections, including Vancomycin-Resistant Staphylococcus aureus (VRSA) and Methicillin-Resistant Staphylococcus aureus (MRSA), using the targeted spectrum of activity of these microorganisms. Method: This study aimed to examine the efficiency of antibiotics and bacteriocin against VRSA and MRSA. The effects of bacteriocins, such as enterocin KAE01, enterocin KAE03, enterocin KAE05, and enterocin KAE06 isolated from Enterococcus faecium strains, alone and in combination with vancomycin and methicillin antibiotics were examined. The selection technique utilized the minimum inhibitory concentrations (MICs) against Gram-positive indicator strain ATCC 6538 Methicillin-Resistant Staphylococcus aureus (MRSA) and indicator strain KSA 02 Vancomycin-Resistant Staphylococcus aureus (VRSA). Results: We report the isolation and identification of enterocins KAE01, KAE03, KAE05, and KAE06 from food isolates of Enterococcus faecium (KAE01, KAE03, KAE05, and KAE06). After isolating the protein, it was partially purified with ammonium sulphate precipitation and purified with fast protein liquid chromatography (FPLC) procedures. Combinations of enterocin KAE01, 1 citric acid, 1 lactic acid, and microcin J25, 1 reuterin, 1 citric acid, and microcin J25, 1 reuterin, 1 lactic acid shown synergistic benefits (FIC index = 0.5) against Vancomycin-Resistant Staphylococcus aureus (VRSA). In addition, a moderately synergistic (FIC index = 0.75) interaction was seen between pediocin PA-1, 1 citric acid, 1 lactic acid, and reuterin 1 citric acid, 1 lactic acid against L. ivanovii HPB28. In the presence of acids, nisin Z exhibited a modestly synergistic effect (FIC index = 0.625-0.75); however, it exhibited additive effects (FIC index = 1) when combined with reuterin or pediocin PA-1 against L. ivanovii HPB28. The efficacy of synergistic consortiums against Gram-positive bacteria was examined. Conclusion: Combining antimicrobials with various modes of action boosted efficacy and expanded the spectrum of inhibition, particularly against multidrug-resistant pathogens, according to our research.

Keywords: Enterococcus faecium, bacteriocin, antimicrobial resistance, antagonistic activity, vancomycin-resistant Staphylococcus aureus, methicillin-resistant Staphylococcus aureus

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Authors: Maliheh Raji, Hossein Abolghasemi, Jaber Safdari, Ali Kargari

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Dysprosium is a rare earth element which is essential for many growing high-technology applications. Dysprosium along with neodymium plays a significant role in different applications such as metal halide lamps, permanent magnets, and nuclear reactor control rods preparation. The purification and separation of rare earth elements are challenging because of their similar chemical and physical properties. Among the various methods, membrane processes provide many advantages over the conventional separation processes such as ion exchange and solvent extraction. In this work, selective extraction and separation of dysprosium from aqueous solutions containing an equimolar mixture of dysprosium and neodymium by emulsion liquid membrane (ELM) was investigated. The organic membrane phase of the ELM was a nanofluid consisting of multiwalled carbon nanotubes (MWCNT), Span80 as surfactant, Cyanex 272 as carrier, kerosene as base fluid, and nitric acid solution as internal aqueous phase. Factors affecting separation of dysprosium such as carrier concentration, MWCNT concentration, feed phase pH and stripping phase concentration were analyzed using Taguchi method. Optimal experimental condition was obtained using analysis of variance (ANOVA) after 10 min extraction. Based on the results, using MWCNT nanofluid in ELM process leads to increase the extraction due to higher stability of membrane and mass transfer enhancement and separation factor of 6 for dysprosium over neodymium can be achieved under the optimum conditions. Additionally, demulsification process was successfully performed and the membrane phase reused effectively in the optimum condition.

Keywords: emulsion liquid membrane, MWCNT nanofluid, separation, Taguchi method

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Authors: Hatice Akbıyık, Hülya Gökmen Özel, Nagehan Emiralioğlu, Elmas Ebru Güneş Yalçın, Deniz Doğru Ersöz, Hayriye Uğur Özçelik, Nural Kiper

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Background: Dietary recommendations in cystic fibrosis (CF) are based on the need to compensate for the increased energy needs of infection, the increased energy cost of breathing and the losses, incurred from malabsorption. Studies in CF indicate that dietary recommendations for CF patients can be difficult to achieve Aim: The aim of this study was to evaluate the energy and nutrient intakes and to compare in accordance with CF dietary guidelines in CF. Methods: One-hundred sixty patients with CF, aged between 2 to 20 years (mean±SD= 7.4±4.8 years) attending Hacettepe University, Faculty of Medicine, Department of Pediatric Pulmonary Diseases were included. Energy and nutrient intakes from foods and enteral products were calculated using a-24-hour dietary recall method with BEBIS 7.2 programme. Percentages of energy and nutrient intakes were compared in accordance with CF dietary guidelines. Patients or/and parents completed a questionnaire showing mealtime problems, usage of alternative therapies and type of nutrition. Statistical analyses were done using SPSS 16.0 programme. Results: It was obtained that 14.5% and 46.9% of the total energy intake were from proteins and carbohydrates, respectively. The actual contribution of total, saturated, monounsaturated and polyunsaturated fats to the total caloric intake was 37.5%, 14.3%, 14.9%, 9.9%, respectively. It was found that 87.7% of energy, 85% of protein 91.7% of carbohydrate, 81.1% of fat intakes were met, when compared CF recommended intakes of 120% RDA. Additionally 67%, 69.5%, 68.2% and 68.9% of the subjects did not achieve CF recommended intakes of 120% RDA for energy, protein, carbohydrate and fat, respectively. Patients with CF had low intakes for age for almost all vitamins and minerals, although supplementation was given. Especially most patients did not achieve the minimum recommended vitamin K intake of 120% RDA. The percentage meeting 120% RDA was 75.9% for vitamin K. It was shown that 41% of the patients had mealtime problems and they skipped the breakfast. Moreover 25.4% of the patients used alternative products outside the standard treatment (such as omega-3, ginger, turmeric, local honey). It was also showed that 60.8% of patients were using enteral products in addition to normal foods, the remaining patients were on only normal foods. Conclusion: The aims of improving nutritional status in children are to achieve normal weight gain and growth; optimize vitamin and mineral status; and slow the rate of clinical decline. In this study although enteral products were used in patients with CF, it was found that energy and nutrient requirements were unable to meet. Because dietary assessment is essential to identify the need for earlier nutritional intervention, in each visit patients need to be referred to CF specialist dietitian.

Keywords: cystic fibrosis, energy and nutrient intakes, mealtime problems, malabsorbtion

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Authors: Gaweł Magdalena, Lipkowska Anna, Olbert Magdalena, Frąckiewicz Ewelina, Librowski Tadeusz, Nowak Gabriel, Pilc Andrzej

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Heme oxygenase-1 (HO-1), also known as heat shock protein 32 (HSP32), has been shown to be implicated in cytoprotection in various organs. Its activation plays a significant role in acute and chronic inflammation, protecting cells from oxidative injury and apoptosis. This inducible isoform of HO catalyzes the first and rate-limiting step in heme degradation to produce equimolar quantities of biologically active products: carbon monoxide (CO), free iron and biliverdin. CO has been reported to possess anti-apoptotic properties. Moreover, it inhibits the production of proinflammatory cytokines and stimulates the synthesis of the anti-inflammatory interleukin-10 (IL-10), as well as promotes vasodilatation at sites of inflammation. The second product of catalytic HO-1 activity, free cytotoxic iron, is promptly sequestered into the iron storage protein ferritin, which lowers the pro-oxidant state of the cell. The third product, biliverdin, is subsequently converted by biliverdin reductase into the bile pigment bilirubin, the most potent endogenous antioxidant among the constituents of human serum, which modulates immune effector functions and suppresses inflammatory response. Furthermore, being one of the so-called stress proteins, HO-1 adaptively responds to different stressors, such as reactive oxygen species (ROS), inflammatory cytokines and heavy metals and thus protects cells against such conditions as ischemia, hemorrhagic shock, heat shock or hypoxia. It is suggested that pharmacologic modulation of HO-1 may represent an effective strategy for prevention of stress and drug-induced gastrointestinal toxicity. HO-1 is constitutively expressed in normal gastric, intestinal and colonic mucosa and up-regulated during inflammation. It has been proven that HO-1 up-regulated by hemin, heme and cobalt-protoporphyrin ameliorates experimental colitis. In addition, the up-regulation of HO-1 partially explains the mechanism of action of 5-aminosalicylic acid (5-ASA), which is used clinically as an anti-colitis agent. In 2009 Ueda et al. has reported for the first time that mucosal protection by Polaprezinc, a chelate compound of zinc and L-carnosine used as an anti-ulcer drug in Japan, is also attributed to induction of HO-1 in the stomach. Since then, inducers of HO-1 are desired subject of research, as they may constitute therapeutically effective anti-ulcer drugs.

Keywords: heme oxygenase-1, gastric lesions, gastroprotection, Polaprezinc

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Authors: N. C. Okoronkwo, I. E. Mbaeyi-Nwaoha, C. P. Agbata

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Childhood malnutrition is one of the most persistent public health problems throughout developing countries, including Nigeria. Demographic and Health survey data from twenty-one developing countries indicated that poor complementary feeding of children aged 6- 23 months contributes to negative growth trends. To reduce malnutrition among children in the society, formulation of complimentary food rich in essential nutrient for optimum growth and development of infants is essential. This study focused on the evaluation of complementary food produced by solid-state fermentation of Acha and Soybean using Rhizopus oligosporus (2710) and Orange-fleshed sweet potatoes (OFSP) using Lactobacillus planterum (B-41621). The raw materials were soaked separately, each in four volumes of 0.9M acetic acid for 16 hours, rinsed with clean water, steam cooked and cooled. Solid-state fermentation (SSF) was carried out by inoculating Acha and Soybean with spore suspension (1x 10⁶spores/ml) of Rhizopus oligosporus (2710) and OFSP with spore suspension (1x 106spores/ml) of Lactobacillus planterum (B-41621). Fermentation which lasted for 72hours was carried out with 24hours sampling. The samples were blended in the following ratios: Acha and soybean 100: 100 (AS), Acha/soybean and OFSP 50: 50(ASO), made into gruel and compared with a commercial infant formula (Cerelac) which served as the control (CTRL). The samples were analyzed for proximate composition using AOAC methods and sensory attributes using a hedonic scale. Results showed that moisture, crude protein, fibre and ash content increased significantly (p<0.05) as fermentation progressed, while carbohydrate and fat content decreased. The protein, moisture, fibre and ash content ranged from 17.10-19.02%, 54.97-56.27%, 7.08-7.60% and2.09-2.38%, respectively, while carbohydrate and fat content ranged from 12.95-10.21% and 5.81-4.52%, respectively. In sensory scores, there were no significant (p>0.05) difference between the average mean scores of colours, texture and consistency of the samples. The sensory score for the overall acceptability ranged from 6.20-7.80. Sample CTRL had the highest score, while sample ASO had the least score. There was no significant (p>0.05) difference between samples CTRL and AS. Solid-state fermentation improved the nutritional content and flavour of the developed complementary food, which is needed for infant growth and development.

Keywords: Complementary food, malnutrition, proximate composition, solid-state fermentation

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Authors: Abdulnasir Hossen, Alaa Barhoum, Deepali Jaju, V. Gowri, L. Al-Kharusi, M. Hassan, K. Al-Hashmi

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Preeclampsia is a pregnancy disorder associated with increase in blood pressure and excess amount of protein in the urine. HRV analysis has been used by many researchers to identify preeclamptic pregnancy from normal pregnancy. A study in this regard to identify preeclamptic pregnancy in Oman from normal pregnant was conducted on 40 subjects (20 patients and 20 normal). The subjects were collected from two hospitals in Oman. A Fast Fourier transform (FFT) spectral analysis has shown that patients with preeclamptic pregnancy have a reduction in the power of the HF band and an increase in the power of the LF band of HRV compared with subjects with normal pregnancy. The accuracy of identification obtained was 80%.

Keywords: preelampsia, pregnancy hypertension, normal pregnant, FFT, spectral analysis, HRV

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Authors: D. Falliano, D. De Domenico, G. Ricciardi, E. Gugliandolo

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This paper is focused on the mechanical characterization of foamed concrete specimens with protein-based foaming agent. Unlike classic foamed concrete, a peculiar property of the analyzed foamed concrete is the extrudability, which is achieved via a specific additive in the concrete mix that significantly improves the cohesion and viscosity of the fresh cementitious paste. A broad experimental campaign was conducted to evaluate the compressive strength and the indirect tensile strength of the specimens. The study has comprised three different cement types, two water/cement ratios, three curing conditions and three target dry densities. The variability of the strength values upon the above mentioned factors is discussed.

Keywords: cement type, curing conditions, density, extrudable concrete, foamed concrete, mechanical characterization

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Authors: Adebola Ajayi

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Ocimum gratissimum belongs to the Lamiaceae family and is know generally as African Basil. Ocimum gratissimum leaves are widely used as local condiments in diets. The leaves were destalked sorted, washed with potable water to remove dirts, air dried for 14 days under ambient temperature and milled into powder. The proximate composition and mineral contents of Ocimum gratissimum leaves were investigated. The proximate analysis showed the moisture, crude, protein, total ash, crude fiber, crude lipid and total carbohydrate contents were 10.72±0.01%, 12.98±0.10%, 10.95±0.42, 10.21±0.04%, 4.81±0.04% and 49.01±0.25% respectively. The results of the analysis showed that Ocimum gratissimum could be a good source of important food nutrients.

Keywords: African Basil, drying, Ocimum gratissimum, proximate

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Authors: Abhinav Mane, Kedar Karvande, Shubham Patel, Abhayraj Lodha

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Gravity filters are one of the most commonly used, economically viable and moderately efficient water purification systems. Their efficiency is mainly based on the type of filter media installed and its location within the filter mass. Several researchers provide valuable input in decision of the type of filter media. However, the choice is mainly restricted to the chemical combinations of different substances. This makes it very much dependent on the factory made filter media, and no cheap alternatives could be found and used. This paper presents the use of disinfectants and filter medias either available naturally or could be prepared using natural resources in conventional mechanism of gravity filter. A small scale laboratory investigation was made with variation in filter media thickness and its location from the top surface of the filter. A rigid steel frame based custom fabricated test setup was used to facilitate placement of filter media at different height within the filter mass. Finely grinded sun dried Neem (Azadirachta indica) extracts and porous burnt clay pads were used as two distinct filter media and placed in isolation as well as in combination with each other. Ground water available in Marathwada region of Maharashtra, India which mainly consists of harmful materials like Arsenic, Chlorides, Iron, Magnesium and Manganese, etc. was treated in the filters fabricated in the present study. The evaluation was made mainly in terms of the input/output water quality assessment through laboratory tests. The present paper should give a cheap and eco-friendly solution to prepare gravity filter at the merit of household skills and availability.

Keywords: fliter media, gravity filters, natural disinfectants, porous clay pads

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Authors: Mahmoodreza Tahamtan

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Malignant surface epithelial ovarian tumors(SEOT) account for approximately 90% of primary ovarian cancer. We evaluate the immunohistochemical expression of WT1 protein among different histologic subtypes of SEOT. Immunohistochemistry for WT1 was done on 35 serous cystadenocarcinomas, 9 borderline serous tumors. A tumor was considered negative if < 1% of tumor cells were stained.Positive reactions were graded as follows:1+,1%-24%; 2+,25%-49%; 3+,50%-74%; 4+,75%-100%. Of the 35 cases of ovarian serous cystadenocarcinoma 30(85.7%)were diffusely positive(3+,4+),4 showed reactivity of < 50% of the tumor cells(1+,2+) and one were negative. All 9 borderline serous tumors showed immunoreactivity with WT1. WT1 is a good marker to distinguish primary ovarian serous carcinomas from other surface epithelial tumors.

Keywords: WT1, ovary, malignant, epithelial tumors

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Authors: Sara Seddiki, Maazouzi Abdelhak

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The main objective of this modest work is to follow the physicochemical and bacteriological evolution of the wastewater from the town of Bechar subjected to purification by filtration according to various local supports, namely Sable and Terrill by reducing nuisances that undergo the receiving environment (Oued Bechar) and therefore make this water source reusable in different areas. The study first made it possible to characterize the urban wastewater of the Bechar wadi, which presents an environmental threat, thus allowing an estimation of the pollutant load, the chemical oxygen demand COD (145 mg / l) and the biological oxygen demand BOD5 (72 mg / l) revealed that these waters are less biodegradable (COD / BOD5 ratio = 0.62), have a fairly high conductivity (2.76 mS/cm), and high levels of mineral matter presented by chlorides and sulphates 390 and 596.1 mg / l respectively, with a pH of 8.1. The characterization of the sand dune (Beni Abbes) shows that quartz (97%) is the most present mineral. The granular analysis allowed us to determine certain parameters like the uniformity coefficient (CU) and the equivalent diameter, and scanning electron microscope (SEM) observations and X-ray analysis were performed. The study of filtered wastewater shows satisfactory and very encouraging treatment results, with complete elimination of total coliforms and streptococci and a good reduction of total aerobic germs in the sand and clay-sand filter. A good yield has been reported in the sand Terrill filter for the reduction of turbidity. The rates of reduction of organic matter in terms of the biological oxygen demand, in chemical oxygen demand recorded, are of the order of 60%. The elimination of sulphates is 40% for the sand filter.

Keywords: urban wastewater, filtration, bacteriological and physicochemical parameters, sand, Terrill, Oued Bechar

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Authors: Thuy Phuong Nhat Tran, Ashutosh Thakur, Toshiaki Taniike

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Recently, covalently linked graphene oxide frameworks (GOFs) have attracted considerable attention in gas absorbance and water purification as well-defined microporous materials. In spite of their potential advantages such as a controllable pore dimension, adjustable hydrophobicity, and structural stability, these materials have been scarcely employed in heterogeneous catalysis. Here we demonstrate a novel and facile method to synthesize Pd nanoparticles (NPs) confined in a GOF (Pd@GOF). The GOF with uniform interlayer space was obtained by the intercalation of diboronic acid between graphene oxide layers. It was found that Pd NPs were generated inside the graphitic gallery spaces of the GOF, and thus, formed Pd NPs were well-dispersed with a narrow particle size distribution. The synthesized Pd@GOF emerged as an efficient nanocatalyst based on its superior performance (product yield and recyclability) toward Suzuki-Miyaura cross-coupling reaction in both polar and apolar solvents, which has been hardly observed for previously reported graphene-based Pd nanocatalysts. Furthermore, the rational comparison of the catalytic performance between two kinds of Pd@GOF (Pd NPs encapsulated in a diboronic ester-intercalated GOF and in a monoboronic ester-intercalated GOF) firmly confirmed the essential role of a rigid framework design in the stabilization of Pd NPs. Based on these results, the covalently assembled GOF was proposed as a promising scaffold for hosting noble metal NPs to construct desired metal@GOF nanocatalysts with improved activity and durability.

Keywords: graphene oxide framework, palladium nanocatalyst, pore confinement, Suzuki-Miyaura cross-coupling reaction

Procedia PDF Downloads 135

Authors: Norah Al Hokbany, Ibrahim Al Jammaz, Basem Al Otaibi, Yousif Al Malki, Subhani M. Okarvi

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Objective: The folate receptor is over-expressed in a wide variety of human tumors. Conjugates of folate have been shown to be selectively taken up by tumor cells via the folate receptor. In an attempt to develop new folate radiotracers with favorable biochemical properties for detecting folate receptor-positive cancers. Methods: we synthesized ⁶⁴Cu-NOTA- and ⁶⁴Cu-NOTAM-folate conjugates using a straightforward and simple one-step reaction. Radiochemical yields were greater than 95% (decay-corrected) with a total synthesis time of less than 20 min. Results: Radiochemical purities were always greater than 98% without high-performance liquid chromatography (HPLC) purification. These synthetic approaches hold considerable promise as a rapid and simple method for ⁶⁴Cu-folate conjugate preparation with high radiochemical yield in a short synthesis time. In vitro tests on the KB cell line showed that significant amounts of the radio conjugates were associated with cell fractions. Bio-distribution studies in nude mice bearing human KB xenografts demonstrated a significant tumor uptake and favorable bio-distribution profile for ⁶⁴Cu-NOTA- and ⁶⁴Cu-NOTAM-folate conjugate. The uptake in the tumors was blocked by the excess injection of folic acid, suggesting a receptor-mediated process. Conclusion: These results demonstrate that the ⁶⁴Cu-NOTAM-folate conjugate may be useful as a molecular probe for the detection and staging of folate receptor-positive cancers, such as ovarian cancer and their metastasis, as well as monitoring tumor response to treatment.

Keywords: folate, receptor, tumor imaging, ⁶⁴Cu-NOTA-folate, PET

Procedia PDF Downloads 106

Authors: A. U. Adamu, E. D Paul, C. E. Gimba, I. G. Ndukwe

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The phytochemical and proximate composition of Aspillia kotschyi belonging to Compositae family which is commonly used as medicinal plant in Nigeria was determined on both the Methanolic and Petroleum sprit extract of the plant. The Methanolic extract of the plant revealed the presence of carbohydrates, cardiac glyscosides, flavonoids, triterpene, and alkaloids. The Petroleum sprit extract showed the presence of only carbohydrates and alkaloid. Proximate composition analysis shows moisture content of 5.7%, total ash of 4.03%, crude protein 10.94%, fibre 9.06%, fat value 0.83%, and nitrogen free extract of 70.19%. The results of this study suggest some merit in the popular use of Aspillia kotschi in herbal medicine.

Keywords: Aspillia kotschyi, herbal medicine, phytochemical, proximate composition

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Authors: Mads H. Clausen

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Plant cell walls are structurally complex and contain a larger number of diverse carbohydrate polymers. These plant fibers are a highly valuable bio-resource and the focus of food, energy and health research. We are interested in studying the interplay of plant cell wall carbohydrates with proteins such as enzymes, cell surface lectins and antibodies. However, detailed molecular level investigations of such interactions are hampered by the heterogeneity and diversity of the polymers of interest. To circumvent this, we target well-defined oligosaccharides with representative structures that can be used for characterizing protein-carbohydrate binding. The presentation will highlight chemical syntheses of plant cell wall oligosaccharides from our group and provide examples from studies of their interactions with proteins.

Keywords: oligosaccharides, carbohydrate chemistry, plant cell walls, carbohydrate-acting enzymes

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Authors: Henry O. Chibudike, Olubamike A. Adeyoju, Bolanle O. Oluwole, Kayode O. Adebowale, Bamidele I. Olu-Owolabi, Chinedum E. Chibudike

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Studies on the Mineral Content of Solojo Flour and Protein Isolates from the two varieties (DAS and BS) of Nigeria cultivated solojo cowpeas were conducted to determine their nutritional value. These inorganic elements or minerals were classified into 3 categories: the ultra-trace minerals, which are the third category; the microelements, also known as the trace minerals, in the second category; while the first category is the macro elements, also known as major minerals. Some of the macro-elements are Ca, P, Na and Cl; the second category, micro-elements include iron, copper, cobalt, potassium, magnesium, iodine, zinc, manganese, molybdenum, F, Cr, Se and S. Results show that the proportion of Sodium (Na) which is ingested into the body in the form of NaCl through food intake maintenance of body pH and to retain water ranged from 728.97 to 253.37 ppm (72.90 to 25.34 mg/100 g); 715.24 to 235.45 ppm; 735.28 to 270.37 ppm; 726.59 to 264.35ppm, for FFDAS, FFBS, DAS and BS respectively with all values of the germinated samples all bellow the control. While FFDAS iron content ranged from 4.25 to 13.50 mg/100 g; FFBS ranged from 3.15 to 12.56 mg/100 g; DAS ranged from 3.81 to 12.90 mg/100g; BS ranged from 3.42 to 9.40 mg/100 g. The values of the germinated flours were all greater than the ungerminated flour. Iron helps to transport oxygen round the body and also helps in red blood cells building and to convert food into needed energy by the body. While Manganese an element that is needed in micro quantity but necessary to convert food into energy, is also crucial for healthy bone and cartilage creation. Results also show that zinc quantity increased as germination proceeded, and the values ranged from 38.80 ppm to 230.00 ppm (3.880 mg/100 g to 23.00 mg/100 g; 0.003880% to 0.0230%); 40.84 to 250.01 ppm; 32.85 to 93.41 ppm; 37.07 to 115.00 ppm, for FFDAS, FFBS, DAS and BS respectively. The Ca content improved significantly (p<0.05) with sprouting; the value extended from 250.56 ppm to 760.03 ppm (25.056 to 76.00 mg/100g or 0.0251 to 0.0760 %); 400.40 to 998.22 ppm; 116.87 to 195.69 ppm; 113.48 to 220.75 ppm, for FFDAS, FFBS, DAS and BS respectively. Zinc element although needed at the micro level in the body, is essential for a strong immune system to keep the body in good health. It is also crucial for the maintenance of a healthy sense of taste and odor, while Calcium is critical for strong bones and teeth, blood coagulation, and muscle tightening and relaxation. Magnesium is needed to build enzymes and antioxidants and also for healthy bones, while Potassium is needed to maintain water balance, muscle movement, and nerve impulses. It functions in conjunction with Na to regulate blood pressure.

Keywords: Solojo cowpea, underutilized legumes, protein isolates, BS, DAS, ungerminated

Procedia PDF Downloads 56

Authors: Indu Gaur, Neha Bhadauria, Shilpi Shilpi, Susmita Goswami, Prem D. Sharma, Prabir K. Paul

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The concept of organic agriculture has been accepted as novelty in Indian society, but there is no data available on the human pathogens colonizing plant parts due to such practices. Also, the pattern and mechanism of their colonization need to be understood in order to devise possible strategies for their prevention. In the present study, human pathogenic bacteria were isolated from organically grown tomato plants and five of them were identified as Klebsiella pneumoniae, Enterobacter ludwigii, Serratia fonticola, Stenotrophomonas maltophilia and Chryseobacterium jejuense. Tomato plants were grown in controlled aseptic conditions with 25±1˚C, 70% humidity and 12 hour L/D photoperiod. Six weeks old plants were divided into 6 groups of 25 plants each and treated as follows: Group 1: K. pneumonia, Group 2: E. ludwigii, Group 3: S. fonticola, Group 4: S. maltophilia, Group 5: C. jejuense, Group 6: Sterile distilled water (control). The inoculums for all treatments were prepared by overnight growth with uniform concentration of 108 cells/ml. Leaf samples from above groups were collected at 0.5, 2, 4, 6 and 24 hours post inoculation for the colony forming unit counts (CFU/cm2 of leaf area) of individual pathogens using leaf impression method. These CFU counts were used for the in vivo colonization assay and adherence assay of individual pathogens. Also, resistance of these pathogens to at least 12 antibiotics was studied. Based on these findings S. fonticola was found to be most prominently colonizing the phylloplane of tomato and was further studied. Tomato plants grown in controlled aseptic conditions same as mentioned above were divided into 2 groups of 25 plants each and treated as follows: Group 1: S. fonticola, Group 2: Sterile distilled water (control). Leaf samples from above groups were collected at 0, 24, 48, 72 and 96 hours post inoculation and homogenized in suitable buffers for surface and cell wall protein isolation. Protein samples thus obtained were subjected to isocratic SDS-gel electrophoresis and analyzed. It was observed that presence of S. fonticola could induce the expression of at least 3 additional cell wall proteins at different time intervals. Surface proteins also showed variation in the expression pattern at different sampling intervals. Further identification of these proteins by MALDI-MS and bioinformatics tools revealed the gene(s) involved in the interaction of S. fonticola with tomato phylloplane.

Keywords: cell wall proteins, human pathogenic bacteria, phylloplane, solanum lycopersicum

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