Search results for: ferric sulphate
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 290

Search results for: ferric sulphate

20 The Effects of in vitro Digestion on Cheese Bioactivity; Comparing Adult and Elderly Simulated in vitro Gastrointestinal Digestion Models

Authors: A. M. Plante, F. O’Halloran, A. L. McCarthy

Abstract:

By 2050 it is projected that 2 billion of the global population will be more than 60 years old. Older adults have unique dietary requirements and aging is associated with physiological changes that affect appetite, sensory perception, metabolism, and digestion. Therefore, it is essential that foods recommended and designed for older adults promote healthy aging. To assess cheese as a functional food for the elderly, a range of commercial cheese products were selected and compared for their antioxidant properties. Cheese from various milk sources (bovine, goats, sheep) with different textures and fat content, including cheddar, feta, goats, brie, roquefort, halloumi, wensleydale and gouda, were initially digested with two different simulated in vitro gastrointestinal digestion (SGID) models. One SGID model represented a validated in vitro adult digestion system and the second model, an elderly SGID, was designed to consider the physiological changes associated with aging. The antioxidant potential of all cheese digestates was investigated using in vitro chemical-based antioxidant assays, (2,2-Diphenyl-1-picrylhydrazyl (DPPH) radical scavenging, ferric reducing antioxidant power (FRAP) and total phenolic content (TPC)). All adult model digestates had high antioxidant activity across both DPPH ( > 70%) and FRAP ( > 700 µM Fe²⁺/kg.fw) assays. Following in vitro digestion using the elderly SGID model, full-fat red cheddar, low-fat white cheddar, roquefort, halloumi, wensleydale, and gouda digestates had significantly lower (p ≤ 0.05) DPPH radical scavenging properties compared to the adult model digestates. Full-fat white cheddar had higher DPPH radical scavenging activity following elderly SGID digestion compared to the adult model digestate, but the difference was not significant. All other cheese digestates from the elderly model were comparable to the digestates from the adult model in terms of radical scavenging activity. The FRAP of all elderly digestates were significantly lower (p ≤ 0.05) compared to the adult digestates. Goats cheese was significantly higher (p ≤ 0.05) in FRAP (718 µM Fe²/kg.fw) compared to all other digestates in the elderly model. TPC levels in the soft cheeses (feta, goats) and low-fat cheeses (red cheddar, white cheddar) were significantly lower (p ≤ 0.05) in the elderly digestates compared to the adult digestates. There was no significant difference in TPC levels, between the elderly and adult model for full-fat cheddar (red, white), roquefort, wensleydale, gouda, and brie digestates. Halloumi cheese was the only cheese that was significantly higher in TPC levels following elderly digestion compared to adult digestates. Low fat red cheddar had significantly higher (p ≤ 0.05) TPC levels compared to all other digestates for both adult and elderly digestive systems. Findings from this study demonstrate that aging has an impact on the bioactivity of cheese, as antioxidant activity and TPC levels were lower, following in vitro elderly digestion compared to the adult model. For older adults, soft cheese, particularly goats cheese, was associated with high radical scavenging and reducing power, while roquefort cheese had low antioxidant activity. Also, elderly digestates of halloumi and low-fat red cheddar were associated with high TPC levels. Cheese has potential as a functional food for the elderly, however, bioactivity can vary depending on the cheese matrix. Funding for this research was provided by the RISAM Scholarship Scheme, Cork Institute of Technology, Ireland.

Keywords: antioxidants, cheese, in-vitro digestion, older adults

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19 Application of a Submerged Anaerobic Osmotic Membrane Bioreactor Hybrid System for High-Strength Wastewater Treatment and Phosphorus Recovery

Authors: Ming-Yeh Lu, Shiao-Shing Chen, Saikat Sinha Ray, Hung-Te Hsu

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Recently, anaerobic membrane bioreactors (AnMBRs) has been widely utilized, which combines anaerobic biological treatment process and membrane filtration, that can be present an attractive option for wastewater treatment and water reuse. Conventional AnMBR is having several advantages, such as improving effluent quality, compact space usage, lower sludge yield, without aeration and production of energy. However, the removal of nitrogen and phosphorus in the AnMBR permeate was negligible which become the biggest disadvantage. In recent years, forward osmosis (FO) is an emerging technology that utilizes osmotic pressure as driving force to extract clean water without additional external pressure. The pore size of FO membrane is kindly mentioned the pore size, so nitrogen or phosphorus could effectively improve removal of nitrogen or phosphorus. Anaerobic bioreactor with FO membrane (AnOMBR) can retain the concentrate organic matters and nutrients. However, phosphorus is a non-renewable resource. Due to the high rejection property of FO membrane, the high amount of phosphorus could be recovered from the combination of AnMBR and FO. In this study, development of novel submerged anaerobic osmotic membrane bioreactor integrated with periodic microfiltration (MF) extraction for simultaneous phosphorus and clean water recovery from wastewater was evaluated. A laboratory-scale AnOMBR utilizes cellulose triacetate (CTA) membranes with effective membrane area of 130 cm² was fully submerged into a 5.5 L bioreactor at 30-35℃. Active layer-facing feed stream orientation was utilized, for minimizing fouling and scaling. Additionally, a peristaltic pump was used to circulate draw solution (DS) at a cross flow velocity of 0.7 cm/s. Magnesium sulphate (MgSO₄) solution was used as DS. Microfiltration membrane periodically extracted about 1 L solution when the TDS reaches to 5 g/L to recover phosphorus and simultaneous control the salt accumulation in the bioreactor. During experiment progressed, the average water flux was achieved around 1.6 LMH. The AnOMBR process show greater than 95% removal of soluble chemical oxygen demand (sCOD), nearly 100% of total phosphorous whereas only partial removal of ammonia, and finally average methane production of 0.22 L/g sCOD was obtained. Therefore, AnOMBR system periodically utilizes MF membrane extracted for phosphorus recovery with simultaneous pH adjustment. The overall performance demonstrates that a novel submerged AnOMBR system is having potential for simultaneous wastewater treatment and resource recovery from wastewater, and hence, the new concept of this system can be used to replace for conventional AnMBR in the future.

Keywords: anaerobic treatment, forward osmosis, phosphorus recovery, membrane bioreactor

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18 Biodsorption as an Efficient Technology for the Removal of Phosphate, Nitrate and Sulphate Anions in Industrial Wastewater

Authors: Angel Villabona-Ortíz, Candelaria Tejada-Tovar, Andrea Viera-Devoz

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Wastewater treatment is an issue of vital importance in these times where the impacts of human activities are most evident, which have become essential tasks for the normal functioning of society. However, they put entire ecosystems at risk by time destroying the possibility of sustainable development. Various conventional technologies are used to remove pollutants from water. Agroindustrial waste is the product with the potential to be used as a renewable raw material for the production of energy and chemical products, and their use is beneficial since products with added value are generated from materials that were not used before. Considering the benefits that the use of residual biomass brings, this project proposes the use of agro-industrial residues from corn crops for the production of natural adsorbents whose purpose is aimed at the remediation of contaminated water bodies with large loads of nutrients. The adsorption capacity of two biomaterials obtained from the processing of corn stalks was evaluated by batch system tests. Biochar impregnated with sulfuric acid and thermally activated was synthesized. On the other hand, the cellulose was extracted from the corn stalks and chemically modified with cetyltrimethylammonium chloride in order to quaternize the surface of the adsorbent. The adsorbents obtained were characterized by thermogravimetric analysis (TGA), scanning electron microscopy (SEM), infrared spectrometry with Fourier Transform (FTIR), analysis by Brunauer, Emmett and Teller method (BET) and X-ray Diffraction analysis ( XRD), which showed favorable characteristics for the cellulose extraction process. Higher adsorption capacities of the nutrients were obtained with the use of biochar, with phosphate being the anion with the best removal percentages. The effect of the initial adsorbate concentration was evaluated, with which it was shown that the Freundlich isotherm better describes the adsorption process in most systems. The adsorbent-phosphate / nitrate systems fit better to the Pseudo Primer Order kinetic model, while the adsorbent-sulfate systems showed a better fit to the Pseudo second-order model, which indicates that there are both physical and chemical interactions in the process. Multicomponent adsorption tests revealed that phosphate anions have a higher affinity for both adsorbents. On the other hand, the thermodynamic parameters standard enthalpy (ΔH °) and standard entropy (ΔS °) with negative results indicate the exothermic nature of the process, whereas the ascending values of standard Gibbs free energy (ΔG °). The adsorption process of anions with biocarbon and modified cellulose is spontaneous and exothermic. The use of the evaluated biomateriles is recommended for the treatment of industrial effluents contaminated with sulfate, nitrate and phosphate anions.

Keywords: adsorption, biochar, modified cellulose, corn stalks

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17 Toxic Chemicals from Industries into Pacific Biota. Investigation of Polychlorinated Biphenyls (PCBs), Dioxins (PCDD), Furans (PCDF) and Polybrominated Diphenyls (PBDE No. 47) in Tuna and Shellfish in Kiribati, Solomon Islands and the Fiji Islands

Authors: Waisea Votadroka, Bert Van Bavel

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The most commonly consumed shellfish species produced in the Pacific, shellfish and tuna fish, were investigated for the occurrence of a range of brominated and chlorinated contaminants in order to establish current levels. Polychlorinated biphenyls (PCBs), polybrominated diphenyl ethers (PBDEs) and polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs) were analysed in the muscle of tuna species Katsuwonis pelamis, yellow fin tuna, and shellfish species from the Fiji Islands. The investigation of polychlorinated biphenyls (PCBs), furans (PCDFs) and polybrominated diphenylethers (PBDE No.47) in tuna and shellfish in Kiribati, Solomon Islands and Fiji is necessary due to the lack of research data in the Pacific region. The health risks involved in the consumption of marine foods laced with toxic organo-chlorinated and brominated compounds makes in the analyses of these compounds in marine foods important particularly when Pacific communities rely on these resources as their main diet. The samples were homogenized in a motor with anhydrous sodium sulphate in the ratio of 1:3 (muscle) and 1:4-1:5 (roe and butter). The tuna and shellfish samples were homogenized and freeze dried at the sampling location at the Institute of Applied Science, Fiji. All samples were stored in amber glss jars at -18 ° C until extraction at Orebro University. PCDD/Fs, PCBs and pesticides were all analysed using an Autospec Ultina HRGC/HRMS operating at 10,000 resolutions with EI ionization at 35 eV. All the measurements were performed in the selective ion recording mode (SIR), monitoring the two most abundant ions of the molecular cluster (PCDD/Fs and PCBs). Results indicated that the Fiji Composite sample for Batissa violacea range 0.7-238.6 pg/g lipid; Fiji sample composite Anadara antiquate range 1.6 – 808.6 pg/g lipid; Solomon Islands Katsuwonis Pelamis 7.5-3770.7 pg/g lipid; Solomon Islands Yellow Fin tuna 2.1 -778.4 pg/g lipid; Kiribati Katsuwonis Pelamis 4.8-1410 pg/g lipids. The study has demonstrated that these species are good bio-indicators of the presence of these toxic organic pollutants in edible marine foods. Our results suggest that for pesticides levels, p,p-DDE is the most dominant for all the groups and seems to be highest at 565.48 pg/g lipid in composite Batissa violacea from Fiji. For PBDE no.47 in comparing all samples, the composite Batissa violacea from Fiji had the highest level of 118.20 pg/g lipid. Based upon this study, the contamination levels found in the study species were quite lower compared with levels reported in impacted ecosystems around the world

Keywords: polychlorinated biphenyl, polybrominated diphenylethers, pesticides, organoclorinated pesticides, PBDEs

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16 Antioxidant Potency of Ethanolic Extracts from Selected Aromatic Plants by in vitro Spectrophotometric Analysis

Authors: Tatjana Kadifkova Panovska, Svetlana Kulevanova, Blagica Jovanova

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Biological systems possess the ability to neutralize the excess of reactive oxygen species (ROS) and to protect cells from destructive alterations. However, many pathological conditions (cardiovascular diseases, autoimmune disorders, cancer) are associated with inflammatory processes that generate an excessive amount of reactive oxygen species (ROS) that shift the balance between endogenous antioxidant systems and free oxygen radicals in favor of the latter, leading to oxidative stress. Therefore, an additional source of natural compounds with antioxidant properties that will reduce the amount of ROS in cells is much needed despite their broad utilization; many plant species remain largely unexplored. Therefore, the purpose of the present study is to investigate the antioxidant activity of twenty-five selected medicinal and aromatic plant species. The antioxidant activity of the ethanol extracts was evaluated with in vitro assays: 2,2’-diphenyl-1-pycryl-hydrazyl (DPPH), ferric reducing antioxidant power (FRAP), non-site-specific- (NSSOH) and site-specific hydroxyl radical-2-deoxy-D-ribose degradation (SSOH) assays. The Folin-Ciocalteu method and AlCl3 method were performed to determine total phenolic content (TPC) and total flavonoid content (TFC). All examined plant extracts manifested antioxidant activity to a different extent. Cinnamomum verum J.Presl bark and Ocimum basilicum L. Herba demonstrated strong radical scavenging activity and reducing power with the DPPH and FRAP assay, respectively. Additionally, significant hydroxyl scavenging potential and metal chelating properties were observed using the NSSOH and SSOH assays. Furthermore, significant variations were determined in the total polyphenolic content (TPC) and total flavonoid content (TFC), with Cinnamomum verum and Ocimum basilicum showing the highest amount of total polyphenols. The considerably strong radical scavenging activity, hydroxyl scavenging potential and reducing power for the species mentioned above suggest of a presence of highly bioactive phytochemical compounds, predominantly polyphenols. Since flavonoids are the most abundant group of polyphenols that possess a large number of available reactive OH groups in their structure, it is considered that they are the main contributors to the radical scavenging properties of the examined plant extracts. This observation is supported by the positive correlation between the radical scavenging activity and the total polyphenolic and flavonoid content obtained in the current research. The observations from the current research nominate Cinnamomum verum bark and Ocimum basilicum herba as potential sources of bioactive compounds that could be utilized as antioxidative additives in the food and pharmaceutical industries. Moreover, the present study will help the researchers as basic data for future research in exploiting the hidden potential of these important plants that have not been explored so far.

Keywords: ethanol extracts, radical scavenging activity, reducing power, total polyphenols.

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15 Fabrication of Electrospun Green Fluorescent Protein Nano-Fibers for Biomedical Applications

Authors: Yakup Ulusu, Faruk Ozel, Numan Eczacioglu, Abdurrahman Ozen, Sabriye Acikgoz

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GFP discovered in the mid-1970s, has been used as a marker after replicated genetic study by scientists. In biotechnology, cell, molecular biology, the GFP gene is frequently used as a reporter of expression. In modified forms, it has been used to make biosensors. Many animals have been created that express GFP as an evidence that a gene can be expressed throughout a given organism. Proteins labeled with GFP identified locations are determined. And so, cell connections can be monitored, gene expression can be reported, protein-protein interactions can be observed and signals that create events can be detected. Additionally, monitoring GFP is noninvasive; it can be detected by under UV-light because of simply generating fluorescence. Moreover, GFP is a relatively small and inert molecule, that does not seem to treat any biological processes of interest. The synthesis of GFP has some steps like, to construct the plasmid system, transformation in E. coli, production and purification of protein. GFP carrying plasmid vector pBAD–GFPuv was digested using two different restriction endonuclease enzymes (NheI and Eco RI) and DNA fragment of GFP was gel purified before cloning. The GFP-encoding DNA fragment was ligated into pET28a plasmid using NheI and Eco RI restriction sites. The final plasmid was named pETGFP and DNA sequencing of this plasmid indicated that the hexa histidine-tagged GFP was correctly inserted. Histidine-tagged GFP was expressed in an Escherichia coli BL21 DE3 (pLysE) strain. The strain was transformed with pETGFP plasmid and grown on LuiraBertoni (LB) plates with kanamycin and chloramphenicol selection. E. coli cells were grown up to an optical density (OD 600) of 0.8 and induced by the addition of a final concentration of 1mM isopropyl-thiogalactopyranoside (IPTG) and then grown for additional 4 h. The amino-terminal hexa-histidine-tag facilitated purification of the GFP by using a His Bind affinity chromatography resin (Novagen). Purity of GFP protein was analyzed by a 12 % sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). The concentration of protein was determined by UV absorption at 280 nm (Varian Cary 50 Scan UV/VIS spectrophotometer). Synthesis of GFP-Polymer composite nanofibers was produced by using GFP solution (10mg/mL) and polymer precursor Polyvinylpyrrolidone, (PVP, Mw=1300000) as starting materials and template, respectively. For the fabrication of nanofibers with the different fiber diameter; a sol–gel solution comprising of 0.40, 0.60 and 0.80 g PVP (depending upon the desired fiber diameter) and 100 mg GFP in 10 mL water: ethanol (3:2) mixtures were prepared and then the solution was covered on collecting plate via electro spinning at 10 kV with a feed-rate of 0.25 mL h-1 using Spellman electro spinning system. Results show that GFP-based nano-fiber can be used plenty of biomedical applications such as bio-imaging, bio-mechanic, bio-material and tissue engineering.

Keywords: biomaterial, GFP, nano-fibers, protein expression

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14 Magnetic Solid-Phase Separation of Uranium from Aqueous Solution Using High Capacity Diethylenetriamine Tethered Magnetic Adsorbents

Authors: Amesh P, Suneesh A S, Venkatesan K A

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The magnetic solid-phase extraction is a relatively new method among the other solid-phase extraction techniques for the separating of metal ions from aqueous solutions, such as mine water and groundwater, contaminated wastes, etc. However, the bare magnetic particles (Fe3O4) exhibit poor selectivity due to the absence of target-specific functional groups for sequestering the metal ions. The selectivity of these magnetic particles can be remarkably improved by covalently tethering the task-specific ligands on magnetic surfaces. The magnetic particles offer a number of advantages such as quick phase separation aided by the external magnetic field. As a result, the solid adsorbent can be prepared with the particle size ranging from a few micrometers to the nanometer, which again offers the advantages such as enhanced kinetics of extraction, higher extraction capacity, etc. Conventionally, the magnetite (Fe3O4) particles were prepared by the hydrolysis and co-precipitation of ferrous and ferric salts in aqueous ammonia solution. Since the covalent linking of task-specific functionalities on Fe3O4 was difficult, and it is also susceptible to redox reaction in the presence of acid or alkali, it is necessary to modify the surface of Fe3O4 by silica coating. This silica coating is usually carried out by hydrolysis and condensation of tetraethyl orthosilicate over the surface of magnetite to yield a thin layer of silica-coated magnetite particles. Since the silica-coated magnetite particles amenable for further surface modification, it can be reacted with task-specific functional groups to obtain the functionalized magnetic particles. The surface area exhibited by such magnetic particles usually falls in the range of 50 to 150 m2.g-1, which offer advantage such as quick phase separation, as compared to the other solid-phase extraction systems. In addition, the magnetic (Fe3O4) particles covalently linked on mesoporous silica matrix (MCM-41) and task-specific ligands offer further advantages in terms of extraction kinetics, high stability, longer reusable cycles, and metal extraction capacity, due to the large surface area, ample porosity and enhanced number of functional groups per unit area on these adsorbents. In view of this, the present paper deals with the synthesis of uranium specific diethylenetriamine ligand (DETA) ligand anchored on silica-coated magnetite (Fe-DETA) as well as on magnetic mesoporous silica (MCM-Fe-DETA) and studies on the extraction of uranium from aqueous solution spiked with uranium to mimic the mine water or groundwater contaminated with uranium. The synthesized solid-phase adsorbents were characterized by FT-IR, Raman, TG-DTA, XRD, and SEM. The extraction behavior of uranium on the solid-phase was studied under several conditions like the effect of pH, initial concentration of uranium, rate of extraction and its variation with pH and initial concentration of uranium, effect of interference ions like CO32-, Na+, Fe+2, Ni+2, and Cr+3, etc. The maximum extraction capacity of 233 mg.g-1 was obtained for Fe-DETA, and a huge capacity of 1047 mg.g-1 was obtained for MCM-Fe-DETA. The mechanism of extraction, speciation of uranium, extraction studies, reusability, and the other results obtained in the present study suggests Fe-DETA and MCM-Fe-DETA are the potential candidates for the extraction of uranium from mine water, and groundwater.

Keywords: diethylenetriamine, magnetic mesoporous silica, magnetic solid-phase extraction, uranium extraction, wastewater treatment

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13 Case Report: Opioid Sparing Anaesthesia with Dexmedetomidine in General Surgery

Authors: Shang Yee Chong

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Perioperative pain is a complex mechanism activated by various nociceptive, neuropathic, and inflammatory pathways. Opioids have long been a mainstay for analgesia in this period, even as we are continuously moving towards a multimodal model to improve pain control while minimising side effects. Dexmedetomidine, a potent alpha-2 agonist, is a useful sedative and hypnotic agent. Its use in the intensive care unit has been well described, and it is increasingly an adjunct intraoperatively for its opioid sparing effects and to decrease pain scores. We describe a case of a general surgical patient in whom minimal opioids was required with dexmedetomidine use. The patient was a 61-year-old Indian gentleman with a history of hyperlipidaemia and type 2 diabetes mellitus, presenting with rectal adenocarcinoma detected on colonoscopy. He was scheduled for a robotic ultra-low anterior resection. The patient was induced with intravenous fentanyl 75mcg, propofol 160mg and atracurium 40mg. He was intubated conventionally and mechanically ventilated. Anaesthesia was maintained with inhalational desflurane and anaesthetic depth was measured with the Masimo EEG Sedline brain function monitor. An initial intravenous dexmedetomidine dose (bolus) of 1ug/kg for 10 minutes was given prior to anaesthetic induction and thereafter, an infusion of 0.2-0.4ug/kg/hr to the end of surgery. In addition, a bolus dose of intravenous lignocaine 1.5mg/kg followed by an infusion at 1mg/kg/hr throughout the surgery was administered. A total of 10mmol of magnesium sulphate and intravenous paracetamol 1000mg were also given for analgesia. There were no significant episodes of bradycardia or hypotension. A total of intravenous phenylephrine 650mcg was given throughout to maintain the patient’s mean arterial pressure within 10-15mmHg of baseline. The surgical time lasted for 5 hours and 40minutes. Postoperatively the patient was reversed and extubated successfully. He was alert and comfortable and pain scores were minimal in the immediate post op period in the postoperative recovery unit. Time to first analgesia was 4 hours postoperatively – with paracetamol 1g administered. This was given at 6 hourly intervals strictly for 5 days post surgery, along with celecoxib 200mg BD as prescribed by the surgeon regardless of pain scores. Oral oxycodone was prescribed as a rescue analgesic for pain scores > 3/10, but the patient did not require any dose. Neither was there nausea or vomiting. The patient was discharged on postoperative day 5. This case has reinforced the use of dexmedetomidine as an adjunct in general surgery cases, highlighting its excellent opioid-sparing effects. In the entire patient’s hospital stay, the only dose of opioid he received was 75mcg of fentanyl at the time of anaesthetic induction. The patient suffered no opioid adverse effects such as nausea, vomiting or postoperative ileus, and pain scores varied from 0-2/10. However, intravenous lignocaine infusion was also used in this instance, which would have helped improve pain scores. Paracetamol, lignocaine, and dexmedetomidine is thus an effective, opioid-sparing combination of multi-modal analgesia for major abdominal surgery cases.

Keywords: analgesia, dexmedetomidine, general surgery, opioid sparing

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12 Influence of Cryo-Grinding on Antioxidant Activity and Amount of Free Phenolic Acids, Rutin and Tyrosol in Whole Grain Buckwheat and Pumpkin Seed Cake

Authors: B. Voucko, M. Benkovic, N. Cukelj, S. Drakula, D. Novotni, S. Balbino, D. Curic

Abstract:

Oxidative stress is considered as one of the causes leading to metabolic disorders in humans. Therefore, the ability of antioxidants to inhibit free radical production is their primary role in the human organism. Antioxidants originating from cereals, especially flavonoids and polyphenols, are mostly bound and indigestible. Micronization damages the cell wall which consecutively results in bioactive material to be more accessible in vivo. In order to ensure complete fragmentation, micronization is often combined with high temperatures (e.g., for bran 200°C) which can lead to degradation of bioactive compounds. The innovative non-thermal technology of cryo-milling is an ultra-fine micronization method that uses liquid nitrogen (LN2) at a temperature of 195°C to freeze and cool the sample during milling. Freezing at such low temperatures causes the material to become brittle which ensures the generation of fine particles while preserving the bioactive content of the material. The aim of this research was to determine if production of ultra-fine material with cryo-milling will result in the augmentation of available bioactive compounds of buckwheat and pumpkin seed cake. For that reason, buckwheat and pumpkin seed cake were ground in a ball mill (CryoMill, Retch, Germany) with and without the use of LN2 for 8 minutes, in a 50 mL stainless steel jar containing one grinding ball (Ø 25 mm) at an oscillation frequency of 30 Hz. The cryo-milled samples were cooled with LN2 for 2 minutes prior to milling, followed by the first cycle of milling (4 minutes), intermediary cooling (2 minutes), and finally the second cycle of milling (further 4 minutes). A continuous process of milling was applied to the samples ground without freezing with LN2. Particle size distribution was determined using the Scirocco 2000 dry dispersion unit (Malvern Instruments, UK). Antioxidant activity was determined by 2,2-Diphenyl-1-picrylhydrazyl (DPPH) test and ferric reducing antioxidant power (FRAP) assay, while the total phenol content was determined using the Folin Ciocalteu method, using the ultraviolet-visible spectrophotometer (Specord 50 Plus, Germany). The content of the free phenolic acids, rutin in buckwheat, tyrosol in pumpkin seed cake, was determined with an HPLC-PDA method (Agilent 1200 series, Germany). Cryo-milling resulted in 11 times smaller size of buckwheat particles, and 3 times smaller size of pumpkin seed particles than milling without the use of LN2, but also, a lower uniformity of the particle size distribution. Lack of freezing during milling of pumpkin seed cake caused a formation of agglomerates due to its high-fat content (21 %). Cryo-milling caused augmentation of buckwheat flour antioxidant activity measured by DPPH test (23,9%) and an increase in available rutin content (14,5%). Also, it resulted in an augmentation of the total phenol content (36,9%) and available tyrosol content (12,5%) of pumpkin seed cake. Antioxidant activity measured with the FRAP test, as well as the content of phenolic acids remained unchanged independent of the milling process. The results of this study showed the potential of cryo-milling for complete raw material utilization in the food industry, as well as a tool for extraction of aimed bioactive components.

Keywords: bioactive, ball-mill, buckwheat, cryo-milling, pumpkin seed cake

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11 Mechanical and Durability Characteristics of Roller Compacted Geopolymer Concrete Using Recycled Concrete Aggregate

Authors: Syfur Rahman, Mohammad J. Khattak

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Every year a huge quantity of recycling concrete aggregate (RCA) is generated in the United States of America. Utilization of RCA can solve the storage problem, prevent environmental pollution, and reduce the construction cost. However, due to the overall low strength and durability characteristics of RCA, its usages are limited to a certain area like a landfill, low strength base material, replacement of a few percentages of virgin aggregates in Portland cement concrete, etc. This study focuses on the improvement of the strength and durability characteristics of RCA by introducing the concept of roller-compacted geopolymer concrete. In this research, developed roller-compacted geopolymer concrete (RCGPC) and roller-compacted cement concrete (RCC) mixtures containing 100% recycled concrete aggregate were evaluated and compared. Several selected RCGPC mixtures were investigated to find out the effect of mixture variables, including sodium hydroxide (NaOH) molar concentration, sodium silicate (Na₂SiO₃), to sodium hydroxide (NaOH) ratio on the strength, stiffness and durability characteristics of the developed RCGPC. Sodium hydroxide (NaOH) and sodium silicate (Na₂SiO₃) were mixed in different ratios to synthesize the alkali activator. American Concrete Pavement Association (ACPA) recommended RCC gradation was used with a maximum nominal aggregate size of 19 mm with a 4% fine particle passing 0.075 mm sieve. The mixtures were made using NaOH molar concentration of 8M and 10M along with, Na₂SiO₃ to NaOH ratio of 0 and 1 by mass and 15% class F fly ash. Optimum alkali content and moisture content were determined for each RCGPC and RCC mixtures, respectively, using modified proctor test. Compressive strength, semi-circular bending beam strength, and dynamic modulus test were conducted to evaluate the mechanistic characteristics of both mixtures. To determine the optimum curing conditions for RCGPC, effects of different curing temperature and curing duration on compressive strength were also studied. Sulphate attack and freeze-thaw tests were also carried out to assess the durability properties of the developed mixtures. X-ray diffraction (XRD) was used for morphology and microstructure analysis. From the optimum moisture content results, it was found that RCGPC has high alkali content, which was mainly due to the high absorption capacity of RCA. It was found that the mixtures with Na₂SiO₃ to NaOH ratio of 1 yielded about 60% higher compressive strength than the ratio of 0. Further, the mixtures using 10M NaOH concentrations and alkali ratio of 1 produced about 28 MPa of compressive strength, which was around 33% higher than 8M NaOH mixtures. Similar results were obtained for elastic and dynamic modulus of the mixtures. On the other hand, the semi-circular bending beam strength remained the same for both 8 and 10 molar NaOH geopolymer mixtures. Formation of new geopolymeric compounds and chemical bonds in the newly formed novel RCGPC mixtures were also discovered using XRD analysis. The results of mechanical and durability testing further revealed that RCGPC performed similarly to that of RCC mixtures. Based on the results of mechanical and durability testing, the developed RCGPC mixtures using 100% recycled concrete could be used as a cost-effective solution for the construction of pavement structures.

Keywords: roller compacted concrete, geopolymer concrete, recycled concrete aggregate, concrete pavement, fly ash

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10 Phytochemical Analysis and in vitro Biological Activities of an Ethyl Acetate Extract from the Peel of Punica granatum L. var. Dente di Cavallo

Authors: Silvia Di Giacomo, Marcello Locatelli, Simone Carradori, Francesco Cacciagrano, Chiara Toniolo, Gabriela Mazzanti, Luisa Mannina, Stefania Cesa, Antonella Di Sotto

Abstract:

Hyperglycemia represents the main pathogenic factor in the development of diabetes complications and has been found associated with mitochondrial dysfunction and oxidative stress, which in turn increase cell dysfunction. Therefore, counteract oxidative species appears to be a suitable strategy for preventing the hyperglycemia-induce cell damage and support the pharmacotherapy of diabetes and metabolic diseases. Antidiabetic potential of many food sources has been linked to the presence of polyphenolic metabolites, particularly flavonoids such as quercetin and its glycosylated form rutin. In line with this evidence, in the present study, we assayed the potential anti-hyperglycemic activity of an ethyl acetate extract from the peel of Punica granatum L. var. Dente di Cavallo (PGE), a fruit well known to traditional medicine for the beneficial properties of its edible juice. The effect of the extract on the glucidic metabolism has been evaluated by assessing its ability to inhibit α-amylase and α-glucosidase, two digestive enzymes responsible for the hydrolysis of dietary carbohydrates: their inhibition can delay the carbohydrate digestion and reduce glucose absorption, thus representing an important strategy for the management of hyperglycemia. Also, the PGE ability to block the release of advanced glycated end-products (AGEs), whose accumulation is known to be responsible for diabetic vascular complications, was studied. The iron-reducing and chelating activities, which are the primary mechanisms by which AGE inhibitors stop their metal-catalyzed formation, were evaluated as possible antioxidant mechanisms. At last, the phenolic content of PGE was characterized by chromatographic and spectrophotometric methods. Our results displayed the ability of PGE to inhibit α-amylase enzyme with a similar potency to the positive control: the IC₅₀ values were 52.2 (CL 27.7 - 101.2) µg/ml and 35.6 (CL 22.8 - 55.5) µg/ml for acarbose and PGE, respectively. PGE also inhibited the α-glucosidase enzyme with about a 25 higher potency than the positive controls of acarbose and quercetin. Furthermore, the extract exhibited ferrous and ferric ion chelating ability, with a maximum effect of 82.1% and 80.6% at a concentration of 250 µg/ml respectively, and reducing properties, reaching the maximum effect of 80.5% at a concentration of 10 µg/ml. At last, PGE was found able to inhibit the AGE production (maximum inhibition of 82.2% at the concentration of 1000 µg/ml), although with lower potency with respect to the positive control rutin. The phytochemical analysis of PGE displayed the presence of high levels of total polyphenols, tannins, and flavonoids, among which ellagic acid, gallic acid and catechin were identified. Altogether these data highlight the ability of PGE to control the carbohydrate metabolism at different levels, both by inhibiting the metabolic enzymes and by affecting the AGE formation likely by chelating mechanisms. It is also noteworthy that peel from pomegranate, although being a waste of juice production, can be reviewed as a nutraceutical source. In conclusion, present results suggest the possible role of PGE as a remedy for preventing hyperglycemia complications and encourage further in vivo studies.

Keywords: anti-hyperglycemic activity, antioxidant properties, nutraceuticals, polyphenols, pomegranate

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9 Biodegradable Cross-Linked Composite Hydrogels Enriched with Small Molecule for Osteochondral Regeneration

Authors: Elena I. Oprita, Oana Craciunescu, Rodica Tatia, Teodora Ciucan, Reka Barabas, Orsolya Raduly, Anca Oancea

Abstract:

Healing of osteochondral defects requires repair of the damaged articular cartilage, the underlying subchondral bone and the interface between these tissues (the functional calcified layer). For this purpose, developing a single monophasic scaffold that can regenerate two specific lineages (cartilage and bone) becomes a challenge. The aim of this work was to develop variants of biodegradable cross-linked composite hydrogel based on natural polypeptides (gelatin), polysaccharides components (chondroitin-4-sulphate and hyaluronic acid), in a ratio of 2:0.08:0.02 (w/w/w) and mixed with Si-hydroxyapatite (Si-Hap), in two ratios of 1:1 and 2:1 (w/w). Si-Hap was synthesized and characterized as a better alternative to conventional Hap. Subsequently, both composite hydrogel variants were cross-linked with (N, N-(3-dimethylaminopropyl)-N-ethyl carbodiimide (EDC) and enriched with a small bioactive molecule (icariin). The small molecule icariin (Ica) (C33H40O15) is the main active constituent (flavonoid) of Herba epimedium used in traditional Chinese medicine to cure bone- and cartilage-related disorders. Ica enhances osteogenic and chondrogenic differentiation of bone marrow mesenchymal stem cells (BMSCs), facilitates matrix calcification and increases the specific extracellular matrix (ECM) components synthesis by chondrocytes. Afterward, the composite hydrogels were characterized for their physicochemical properties in terms of the enzymatic biodegradation in the presence of type I collagenase and trypsin, the swelling capacity and the degree of crosslinking (TNBS assay). The cumulative release of Ica and real-time concentration were quantified at predetermined periods of time, according to the standard curve of standard Ica, after hydrogels incubation in saline buffer at physiological parameters. The obtained cross-linked composite hydrogels enriched with small-molecule Ica were also characterized for morphology by scanning electron microscopy (SEM). Their cytocompatibility was evaluated according to EN ISO 10993-5:2009 standard for medical device testing. Thus, analyses regarding cell viability (Live/Dead assay), cell proliferation (Neutral Red assay) and cell adhesion to composite hydrogels (SEM) were performed using NCTC clone L929 cell line. The final results showed that both cross-linked composite hydrogel variants enriched with Ica presented optimal physicochemical, structural and biological properties to be used as a natural scaffold able to repair osteochondral defects. The data did not reveal any toxicity of composite hydrogels in NCTC stabilized cell lines within the tested range of concentrations. Moreover, cells were capable of spreading and proliferating on both composite hydrogel surfaces. In conclusion, the designed biodegradable cross-linked composites enriched with Si and Ica are recommended for further testing as natural temporary scaffolds, which can allow cell migration and synthesis of new extracellular matrix within osteochondral defects.

Keywords: composites, gelatin, osteochondral defect, small molecule

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8 Liposome Loaded Polysaccharide Based Hydrogels: Promising Delayed Release Biomaterials

Authors: J. Desbrieres, M. Popa, C. Peptu, S. Bacaita

Abstract:

Because of their favorable properties (non-toxicity, biodegradability, mucoadhesivity etc.), polysaccharides were studied as biomaterials and as pharmaceutical excipients in drug formulations. These formulations may be produced in a wide variety of forms including hydrogels, hydrogel based particles (or capsules), films etc. In these formulations, the polysaccharide based materials are able to provide local delivery of loaded therapeutic agents but their delivery can be rapid and not easily time-controllable due to, particularly, the burst effect. This leads to a loss in drug efficiency and lifetime. To overcome the consequences of burst effect, systems involving liposomes incorporated into polysaccharide hydrogels may appear as a promising material in tissue engineering, regenerative medicine and drug loading systems. Liposomes are spherical self-closed structures, composed of curved lipid bilayers, which enclose part of the surrounding solvent into their structure. The simplicity of production, their biocompatibility, the size and similar composition of cells, the possibility of size adjustment for specific applications, the ability of hydrophilic or/and hydrophobic drug loading make them a revolutionary tool in nanomedicine and biomedical domain. Drug delivery systems were developed as hydrogels containing chitosan or carboxymethylcellulose (CMC) as polysaccharides and gelatin (GEL) as polypeptide, and phosphatidylcholine or phosphatidylcholine/cholesterol liposomes able to accurately control this delivery, without any burst effect. Hydrogels based on CMC were covalently crosslinked using glutaraldehyde, whereas chitosan based hydrogels were double crosslinked (ionically using sodium tripolyphosphate or sodium sulphate and covalently using glutaraldehyde). It has been proven that the liposome integrity is highly protected during the crosslinking procedure for the formation of the film network. Calcein was used as model active matter for delivery experiments. Multi-Lamellar vesicles (MLV) and Small Uni-Lamellar Vesicles (SUV) were prepared and compared. The liposomes are well distributed throughout the whole area of the film, and the vesicle distribution is equivalent (for both types of liposomes evaluated) on the film surface as well as deeper (100 microns) in the film matrix. An obvious decrease of the burst effect was observed in presence of liposomes as well as a uniform increase of calcein release that continues even at large time scales. Liposomes act as an extra barrier for calcein release. Systems containing MLVs release higher amounts of calcein compared to systems containing SUVs, although these liposomes are more stable in the matrix and diffuse with difficulty. This difference comes from the higher quantity of calcein present within the MLV in relation with their size. Modeling of release kinetics curves was performed and the release of hydrophilic drugs may be described by a multi-scale mechanism characterized by four distinct phases, each of them being characterized by a different kinetics model (Higuchi equation, Korsmeyer-Peppas model etc.). Knowledge of such models will be a very interesting tool for designing new formulations for tissue engineering, regenerative medicine and drug delivery systems.

Keywords: controlled and delayed release, hydrogels, liposomes, polysaccharides

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7 Testicular Differential MicroRNA Expression Derived Occupational Risk Factor Assessment in Idiopathic Non-obstructive Azoospermia Cases

Authors: Nisha Sharma, Mili Kaur, Ashutosh Halder, Seema Kaushal, Manoj Kumar, Manish Jain

Abstract:

Purpose: To investigate microRNAs (miRNA) as an epigenomic etiological factor in idiopathic non-obstructive azoospermia (NOA). In order to achieve the same, an association was seen between occupational exposure to radiation, thermal, and chemical factors and idiopathic cases of non-obstructive azoospermia, and later, testicular differential miRNA expression profiling was done in exposure group NOA cases. Method: It is a prospective study in which 200 apparent idiopathic male factor infertility cases, who have been advised to undergo testicular fine needle aspiration (FNA) evaluation, are recruited. A detailed occupational history was taken to understand the possible type of exposure due to the nature and duration of work. A total of 26 patients were excluded upon XY-FISH and Yq microdeletion tests due to the presence of genetic causes of infertility, 6 hypospermatogeneis (HS), six Sertoli cell-only syndrome (SCOS), and six normospermatogeneis patients testicular FNA samples were used for RNA isolation followed by small RNA sequencing and nCounter miRNA expression analysis. Differential miRNA expression profile of HS and SCOS patients was done. A web-based tool, miRNet, was used to predict the interacting compounds or chemicals using the shortlisted miRNAs with high fold change. The major limitation encountered in this study was the insufficient quantity of testicular FNA sample used for total RNA isolation, which resulted in a low yield and RNA integrity number (RIN) value. Therefore, the number of RNA samples admissible for differential miRNA expression analysis was very small in comparison to the total number of patients recruited. Results: Differential expression analysis revealed 69 down-regulated and 40 up-regulated miRNAs in HS and 66 down-regulated and 33 up-regulated miRNAs in SCOS in comparison to normospermatogenesis controls. The miRNA interaction analysis using the miRNet tool showed that the differential expression profiles of HS and SCOS patients were associated with arsenic trioxide, bisphenol-A, calcium sulphate, lithium, and cadmium. These compounds are reproductive toxins and might be responsible for miRNA-mediated epigenetic deregulation leading to NOA. The association between occupational risk factor exposure and the non-exposure group of NOA patients was not statistically significant, with ꭓ2 (3, N= 178) = 6.70, p= 0.082. The association between individual exposure groups (radiation, thermal, and chemical) and various sub-types of NOA is also not significant, with ꭓ2 (9, N= 178) = 15.06, p= 0.089. Functional analysis of HS and SCOS patients' miRNA profiles revealed some important miR-family members in terms of male fertility. The miR-181 family plays a role in the differentiation of spermatogonia and spermatocytes, as well as the transcriptional regulation of haploid germ cells. The miR-34 family is expressed in spermatocytes and round spermatids and is involved in the regulation of SSCs differentiation. Conclusion: The reproductive toxins might adopt the miRNA-mediated mechanism of disease development in idiopathic cases of NOA. Chemical compound induced; miRNA-mediated epigenetic deregulation can give a future perspective on the etiopathogenesis of the disease.

Keywords: microRNA, non-obstructive azoospermia (NOA), occupational exposure, hypospermatogenesis (HS), Sertoli cell only syndrome (SCOS)

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6 From Biowaste to Biobased Products: Life Cycle Assessment of VALUEWASTE Solution

Authors: Andrés Lara Guillén, José M. Soriano Disla, Gemma Castejón Martínez, David Fernández-Gutiérrez

Abstract:

The worldwide population is exponentially increasing, which causes a rising demand for food, energy and non-renewable resources. These demands must be attended to from a circular economy point of view. Under this approach, the obtention of strategic products from biowaste is crucial for the society to keep the current lifestyle reducing the environmental and social issues linked to the lineal economy. This is the main objective of the VALUEWASTE project. VALUEWASTE is about valorizing urban biowaste into proteins for food and feed and biofertilizers, closing the loop of this waste stream. In order to achieve this objective, the project validates three value chains, which begin with the anaerobic digestion of the biowaste. From the anaerobic digestion, three by-products are obtained: i) methane that is used by microorganisms, which will be transformed into microbial proteins; ii) digestate that is used by black soldier fly, producing insect proteins; and iii) a nutrient-rich effluent, which will be transformed into biofertilizers. VALUEWASTE is an innovative solution, which combines different technologies to valorize entirely the biowaste. However, it is also required to demonstrate that the solution is greener than other traditional technologies (baseline systems). On one hand, the proteins from microorganisms and insects will be compared with other reference protein production systems (gluten, whey and soybean). On the other hand, the biofertilizers will be compared to the production of mineral fertilizers (ammonium sulphate and synthetic struvite). Therefore, the aim of this study is to provide that biowaste valorization can reduce the environmental impacts linked to both traditional proteins manufacturing processes and mineral fertilizers, not only at a pilot-scale but also at an industrial one. In the present study, both baseline system and VALUEWASTE solution are evaluated through the Environmental Life Cycle Assessment (E-LCA). The E-LCA is based on the standards ISO 14040 and 14044. The Environmental Footprint methodology was the one used in this study to evaluate the environmental impacts. The results for the baseline cases show that the food proteins coming from whey have the highest environmental impact on ecosystems compared to the other proteins sources: 7.5 and 15.9 folds higher than soybean and gluten, respectively. Comparing feed soybean and gluten, soybean has an environmental impact on human health 195.1 folds higher. In the case of biofertilizers, synthetic struvite has higher impacts than ammonium sulfate: 15.3 (ecosystems) and 11.8 (human health) fold, respectively. The results shown in the present study will be used as a reference to demonstrate the better environmental performance of the bio-based products obtained through the VALUEWASTE solution. Other originalities that the E-LCA performed in the VALUEWASTE project provides are the diverse direct implications on investment and policies. On one hand, better environmental performance will serve to remove the barriers linked to these kinds of technologies, boosting the investment that is backed by the E-LCA. On the other hand, it will be a germ to design new policies fostering these types of solutions to achieve two of the key targets of the European Community: being self-sustainable and carbon neutral.

Keywords: anaerobic digestion, biofertilizers, circular economy, nutrients recovery

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5 Bioinspired Green Synthesis of Magnetite Nanoparticles Using Room-Temperature Co-Precipitation: A Study of the Effect of Amine Additives on Particle Morphology in Fluidic Systems

Authors: Laura Norfolk, Georgina Zimbitas, Jan Sefcik, Sarah Staniland

Abstract:

Magnetite nanoparticles (MNP) have been an area of increasing research interest due to their extensive applications in industry, such as in carbon capture, water purification, and crucially, the biomedical industry. The use of MNP in the biomedical industry is rising, with studies on their effect as Magnetic resonance imaging contrast agents, drug delivery systems, and as hyperthermic cancer treatments becoming prevalent in the nanomaterial research community. Particles used for biomedical purposes must meet stringent criteria; the particles must have consistent shape and size between particles. Variation between particle morphology can drastically alter the effective surface area of the material, making it difficult to correctly dose particles that are not homogeneous. Particles of defined shape such as octahedral and cubic have been shown to outperform irregular shaped particles in some applications, leading to the need to synthesize particles of defined shape. In nature, highly homogeneous MNP are found within magnetotactic bacteria, a unique bacteria capable of producing magnetite nanoparticles internally under ambient conditions. Biomineralisation proteins control the properties of the MNPs, enhancing their homogeneity. One of these proteins, Mms6, has been successfully isolated and used in vitro as an additive in room-temperature co-precipitation reactions (RTCP) to produce particles of defined mono-dispersed size & morphology. When considering future industrial scale-up it is crucial to consider the costs and feasibility of an additive, as an additive that is not readily available or easily synthesized at a competitive price will not be sustainable. As such, additives selected for this research are inspired by the functional groups of biomineralisation proteins, but cost-effective, environmentally friendly, and compatible with scale-up. Diethylenetriamine (DETA), triethylenetetramine (TETA), tetraethylenepentamine (TEPA), and pentaethylenehexamine (PEHA) have been successfully used in RTCP to modulate the properties of particles synthesized, leading to the formation of octahedral nanoparticles with no use of organic solvents, heating, or toxic precursors. By extending this principle to a fluidic system, ongoing research will reveal whether the amine additives can also exert morphological control in an environment which is suited toward higher particle yield. Two fluidic systems have been employed; a peristaltic turbulent flow mixing system suitable for the rapid production of MNP, and a macrofluidic system for the synthesis of tailored nanomaterials under a laminar flow regime. The presence of the amine additives in the turbulent flow system in initial results appears to offer similar morphological control as observed under RTCP conditions, with higher proportions of octahedral particles formed. This is a proof of concept which may pave the way to green synthesis of tailored MNP on an industrial scale. Mms6 and amine additives have been used in the macrofluidic system, with Mms6 allowing magnetite to be synthesized at unfavourable ferric ratios, but no longer influencing particle size. This suggests this synthetic technique while still benefiting from the addition of additives, may not allow additives to fully influence the particles formed due to the faster timescale of reaction. The amine additives have been tested at various concentrations, the results of which will be discussed in this paper.

Keywords: bioinspired, green synthesis, fluidic, magnetite, morphological control, scale-up

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4 Biomedical Application of Green Biosynthesis Magnetic Iron Oxide (Fe3O4) Nanoparticles Using Seaweed (Sargassum muticum) Aqueous Extract

Authors: Farideh Namvar, Rosfarizan Mohamed

Abstract:

In the field of nanotechnology, the use of various biological units instead of toxic chemicals for the reduction and stabilization of nanoparticles, has received extensive attention. This use of biological entities to create nanoparticles has designated as “Green” synthesis and it is considered to be far more beneficial due to being economical, eco-friendly and applicable for large-scale synthesis as it operates on low pressure, less input of energy and low temperatures. The lack of toxic byproducts and consequent decrease in degradation of the product renders this technique more preferable over physical and classical chemical methods. The variety of biomass having reduction properties to produce nanoparticles makes them an ideal candidate for fabrication. Metal oxide nanoparticles have been said to represent a "fundamental cornerstone of nanoscience and nanotechnology" due to their variety of properties and potential applications. However, this also provides evidence of the fact that metal oxides include many diverse types of nanoparticles with large differences in chemical composition and behaviour. In this study, iron oxide nanoparticles (Fe3O4-NPs) were synthesized using a rapid, single step and completely green biosynthetic method by reduction of ferric chloride solution with brown seaweed (Sargassum muticum) water extract containing polysaccharides as a main factor which acts as reducing agent and efficient stabilizer. Antimicrobial activity against six microorganisms was tested using well diffusion method. The resulting S-IONPs are crystalline in nature, with a cubic shape. The average particle diameter, as determined by TEM, was found to be 18.01 nm. The S-IONPs were efficiently inhibited the growth of Listeria monocytogenes, Escherichia coli and Candida species. Our favorable results suggest that S-IONPs could be a promising candidate for development of future antimicrobial therapies. The nature of biosynthesis and the therapeutic potential by S-IONPs could pave the way for further research on design of green synthesis therapeutic agents, particularly nanomedicine, to deal with treatment of infections. Further studies are needed to fully characterize the toxicity and the mechanisms involved with the antimicrobial activity of these particles. Antioxidant activity of S-IONPs synthesized by green method was measured by ABTS (2, 2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) (IC50= 1000µg) radical scavenging activity. Also, with the increasing concentration of S-IONPs, catalase gene expression compared to control gene GAPDH increased. For anti-angiogenesis study the Ross fertilized eggs were divided into four groups; the control and three experimental groups. The gelatin sponges containing albumin were placed on the chorioalantoic membrane and soaked with different concentrations of S-IONPs. All the cases were photographed using a photo stereomicroscope. The number and the lengths of the vessels were measured using Image J software. The crown rump (CR) and weight of the embryo were also recorded. According to the data analysis, the number and length of the blood vessels, as well as the CR and weight of the embryos reduced significantly compared to the control (p < 0.05), dose dependently. The total hemoglobin was quantified as an indicator of the blood vessel formation, and in the treated samples decreased, which showed its inhibitory effect on angiogenesis.

Keywords: anti-angiogenesis, antimicrobial, antioxidant, biosynthesis, iron oxide (fe3o4) nanoparticles, sargassum muticum, seaweed

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3 Evaluation of Functional Properties of Protein Hydrolysate from the Fresh Water Mussel Lamellidens marginalis for Nutraceutical Therapy

Authors: Jana Chakrabarti, Madhushrita Das, Ankhi Haldar, Roshni Chatterjee, Tanmoy Dey, Pubali Dhar

Abstract:

High incidences of Protein Energy Malnutrition as a consequence of low protein intake are quite prevalent among the children in developing countries. Thus prevention of under-nutrition has emerged as a critical challenge to India’s developmental Planners in recent times. Increase in population over the last decade has led to greater pressure on the existing animal protein sources. But these resources are currently declining due to persistent drought, diseases, natural disasters, high-cost of feed, and low productivity of local breeds and this decline in productivity is most evident in some developing countries. So the need of the hour is to search for efficient utilization of unconventional low-cost animal protein resources. Molluscs, as a group is regarded as under-exploited source of health-benefit molecules. Bivalve is the second largest class of phylum Mollusca. Annual harvests of bivalves for human consumption represent about 5% by weight of the total world harvest of aquatic resources. The freshwater mussel Lamellidens marginalis is widely distributed in ponds and large bodies of perennial waters in the Indian sub-continent and well accepted as food all over India. Moreover, ethno-medicinal uses of the flesh of Lamellidens among the rural people to treat hypertension have been documented. Present investigation thus attempts to evaluate the potential of Lamellidens marginalis as functional food. Mussels were collected from freshwater ponds and brought to the laboratory two days before experimentation for acclimatization in laboratory conditions. Shells were removed and fleshes were preserved at- 20oC until analysis. Tissue homogenate was prepared for proximate studies. Fatty acids and amino acids composition were analyzed. Vitamins, Minerals and Heavy metal contents were also studied. Mussel Protein hydrolysate was prepared using Alcalase 2.4 L and degree of hydrolysis was evaluated to analyze its Functional properties. Ferric Reducing Antioxidant Power (FRAP) and DPPH Antioxidant assays were performed. Anti-hypertensive property was evaluated by measuring Angiotensin Converting Enzyme (ACE) inhibition assay. Proximate analysis indicates that mussel meat contains moderate amount of protein (8.30±0.67%), carbohydrate (8.01±0.38%) and reducing sugar (4.75±0.07%), but less amount of fat (1.02±0.20%). Moisture content is quite high but ash content is very low. Phospholipid content is significantly high (19.43 %). Lipid constitutes, substantial amount of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) which have proven prophylactic values. Trace elements are found present in substantial amount. Comparative study of proximate nutrients between Labeo rohita, Lamellidens and cow’s milk indicates that mussel meat can be used as complementary food source. Functionality analyses of protein hydrolysate show increase in Fat absorption, Emulsification, Foaming capacity and Protein solubility. Progressive anti-oxidant and anti-hypertensive properties have also been documented. Lamellidens marginalis can thus be regarded as a functional food source as this may combine effectively with other food components for providing essential elements to the body. Moreover, mussel protein hydrolysate provides opportunities for utilizing it in various food formulations and pharmaceuticals. The observations presented herein should be viewed as a prelude to what future holds.

Keywords: functional food, functional properties, Lamellidens marginalis, protein hydrolysate

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2 White-Rot Fungi Phellinus as a Source of Antioxidant and Antitumor Agents

Authors: Yogesh Dalvi, Ruby Varghese, Nibu Varghese, C. K. Krishnan Nair

Abstract:

Introduction: The Genus Phellinus, locally known as Phansomba is a well-known traditional folk medicine. Especially, in Western Ghats of India, many tribes use several species of Phellinus for various ailments related to teeth, throat, tongue, stomach and even wound healing. It is one of the few mushrooms which play a pivotal role in Ayurvedic Dravyaguna. Aim: The present study focuses on to investigate phytochemical analysis, antioxidant, and antitumor (in vitro and in vivo) potential of Phellinus robinae from South India, Kerala Material and Methods: The present study explores the following: 1. Phellinus samples were collected from Ranni, Pathanamthitta district of Kerala state, India from Artocarpus heterophyllus Lam. and species were identified using rDNA region. 2. The fruiting body was shadow dried, powdered and extracted with 50% alcohol using water bath at 60°C which was further condensed by rotary evaporator and lyophilized at minus 40°C temperature. 3. Secondary metabolites were analyzed by using various phytochemical screening assay (Hager’s Test, Wagner’s Test, Sodium hydroxide Test, Lead acetate Test, Ferric chloride Test, Folin-ciocalteu Test, Foaming Test, Benedict’s test, Fehling’s Test and Lowry’s Test). 4. Antioxidant and free radical scavenging activity were analyzed by DPPH, FRAP and Iron chelating assay. 5. The antitumor potential of Water alcohol extract of Phellinus (PAWE) is evaluated through In vitro condition by Trypan blue dye exclusion method in DLA cell line and In vivo by murine model. Result and Discussion: Preliminary phytochemical screening by various biochemical tests revealed presence of a variety of active secondary molecules like alkaloids, flavanoids, saponins, carbohydrate, protein and phenol. In DPPH and FRAP assay PAWE showed significantly higher antioxidant activity as compared to standard Ascorbic acid. While, in Iron chelating assay, PAWE exhibits similar antioxidant activity that of Butylated Hydroxytoluene (BHT) as standard. Further, in the in vitro study, PAWE showed significant inhibition on DLA cell proliferation in dose dependent manner and showed no toxicity on mice splenocytes, when compared to standard chemotherapy drug doxorubicin. In vivo study, oral administration of PAWE showed dose dependent tumor regression in mice and also raised the immunogenicity by restoring levels of antioxidant enzymes in liver and kidney tissue. In both in vitro and in vivo gene expression studies PAWE up-regulates pro-apoptotic genes (Bax, Caspases 3, 8 and 9) and down- regulates anti-apoptotic genes (Bcl2). PAWE also down regulates inflammatory gene (Cox-2) and angiogenic gene (VEGF). Conclusion: Preliminary phytochemical screening revealed that PAWE contains various secondary metabolites which contribute to its antioxidant and free radical scavenging property as evaluated by DPPH, FRAP and Iron chelating assay. PAWE exhibits anti-proliferative activity by the induction of apoptosis through a signaling cascade of death receptor-mediated extrinsic (Caspase8 and Tnf-α), as well as mitochondria-mediated intrinsic (caspase9) and caspase pathways (Caspase3, 8 and 9) and also by regressing angiogenic factor (VEGF) without any inflammation or adverse side effects. Hence, PAWE serve as a potential antioxidant and antitumor agent.

Keywords: antioxidant, antitumor, Dalton lymphoma ascites (DLA), fungi, Phellinus robinae

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1 La0.80Ag0.15MnO3 Magnetic Nanoparticles for Self-Controlled Magnetic Fluid Hyperthermia

Authors: Marian Mihalik, Kornel Csach, Martin Kovalik, Matúš Mihalik, Martina Kubovčíková, Maria Zentková, Martin Vavra, Vladimír Girman, Jaroslav Briančin, Marija Perovic, Marija Boškovic, Magdalena Fitta, Robert Pelka

Abstract:

Current nanomaterials for use in biomedicine are based mainly on iron oxides and on present knowledge on magnetic nanostructures. Manganites can represent another material which can be used optionally. Manganites and their unique electronic properties have been extensively studied in the last decades not only due to fundamental interest but to possible applications of colossal magnetoresistance, magnetocaloric effect, and ferroelectric properties. It was found that the oxygen-reduction reaction on perovskite oxide is intimately connected with metal ion e.g., orbital occupation. The effect of oxygen deviation from the stoichiometric composition on crystal structure was studied very carefully by many authors on LaMnO₃. Depending on oxygen content, the crystal structure changes from orthorhombic one to rhombohedric for oxygen content 3.1. In the case of hole-doped manganites, the change from the orthorhombic crystal structure, which is typical for La1-xCaxMnO3 based manganites, to the rhombohedric crystal structure (La1-xMxMnO₃ where M = K, Ag, and Sr based materials) results in an enormous increase of the Curie temperature. In our paper, we study the effect of oxygen content on crystal structure, thermal, and magnetic properties (including magnetocaloric effect) of La1-xAgxMnO₃nano particle system. The content of oxygen in samples was tuned by heat treatment in different thermal regimes and in various environment (air, oxygen, argon). Water nanosuspensions based on La0.80Ag0.15MnO₃ magnetic particles with the Curie temperature of about 43oC were prepared by two different approaches. First, by using a laboratory circulation mill for milling of powder in the presence of sodium dodecyl sulphate (SDS) and subsequent centrifugation. Second nanosuspension was prepared using an agate bowl, etching in citric acid and HNO3, ultrasound homogeniser, centrifugation, and dextran 40 kDA or 15 kDA as surfactant. Electrostatic stabilisation obtained by the first approach did not offer long term kinetic and aggregation colloidal stability and was unable to compensate for attractive forces between particles under a magnetic field. By the second approach, we prepared suspension oversaturated by dextran 40 kDA for steric stabilisation, with evidence of the presence of superparamagnetic behaviour. Low concentration of nanoparticles and not ideal coverage of nanoparticles impacting the stability of ferrofluids was the disadvantage of this approach. Strong steric stabilisation was observable at alcaic conditions under pH = ~10. Application of dextran 15 kDA leads to relatively stable ferrofluid with pH around physiological conditions, but desegregation of powder by HNO₃ was not effective enough, and the average size of fragments was to large of about 150 nm, and we did not see any signature of superparamagnetic behaviour. The prepared ferrofluids were characterised by scanning and transition microscope method, thermogravimetry, magnetization, and AC susceptibility measurements. Specific Absorption Rate measurements were undertaken on powder as well on ferrofluids in order to estimate the potential application of La₀.₈₀Ag₀.₁₅MnO₃ magnetic particles based ferrofluid for hyperthermia. Our complex study contains an investigation of biocompatibility and potential biohazard of this material.

Keywords: manganites, magnetic nanoparticles, oxygen content, magnetic phase transition, magnetocaloric effect, ferrofluid, hyperthermia

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