Search results for: intracellular sodium
799 Soil Quality Status under Dryland Vegetation of Yabello District, Southern Ethiopia
Authors: Mohammed Abaoli, Omer Kara
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The current research has investigated the soil quality status under dryland vegetation of Yabello district, Southern Ethiopia in which we should identify the nature and extent of salinity problem of the area for further research bases. About 48 soil samples were taken from 0-30, 31-60, 61-90 and 91-120 cm soil depths by opening 12 representative soil profile pits at 1.5 m depth. Soil color, texture, bulk density, Soil Organic Carbon (SOC), Cation Exchange Capacity (CEC), Na, K, Mg, Ca, CaCO3, gypsum (CaSO4), pH, Sodium Adsorption Ratio (SAR), Exchangeable Sodium Percentage (ESP) were analyzed. The dominant soil texture was silty-clay-loam. Bulk density varied from 1.1 to 1.31 g/cm3. High SOC content was observed in 0-30 cm. The soil pH ranged from 7.1 to 8.6. The electrical conductivity shows indirect relationship with soil depth while CaCO3 and CaSO4 concentrations were observed in a direct relationship with depth. About 41% are non-saline, 38.31% saline, 15.23% saline-sodic and 5.46% sodic soils. Na concentration in saline soils was greater than Ca and Mg in all the soil depths. Ca and Mg contents were higher above 60 cm soil depth in non-saline soils. The concentrations of SO2-4 and HCO-3 were observed to be higher at the most lower depth than upper. SAR value tends to be higher at lower depths in saline and saline-sodic soils, but decreases at lower depth of the non-saline soils. The distribution of ESP above 60 cm depth was in an increasing order in saline and saline-sodic soils. The result of the research has shown the direction to which extent of salinity we should consider for the Commiphora plant species we want to grow on the area.Keywords: commiphora species, dryland vegetation, ecological significance, soil quality, salinity problem
Procedia PDF Downloads 195798 Preservation of Phenytoin and Sodium Valproate Induced Bone Loss by Raloxifene through Modulating Serum Estradiol and TGF-β3 Content in Bone of Female Mice
Authors: Divya Vohora, Md. Jamir Anwar
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Antiepileptic drugs (AEDs)-induced adverse consequences on bone are now well recognized. Despite this, there is limited data on the effect of anti-osteoporotic therapies on AEDs-induced bone loss. Both phenytoin (PHT) and sodium valproate (SVP) inhibit human aromatase enzyme and stimulate microsomal catabolism of oestrogens. Estrogen deficiency states are known to reduce the deposition of transforming growth factor-β (TGF-β3), a bone matrix protein, having anti-osteoclastic property. Thus, an attempt was made to investigate the effect of raloxifene, a selective oestrogen receptor modulator, in comparison with CVD supplementation, on PHT and SVP-induced alterations in bone in mice. Further, the effect of raloxifene on seizures and on the antiepileptic efficacy of AEDs was also investigated. Swiss strains of female mice were treated with PHT (35 mg/kg, p.o.) and SVP (300 mg/kg, p.o.) for 120 days to induce bone loss as evidenced by reduced bone mineral density (BMD) and altered bone turnover markers in lumbar bones (alkaline phosphatase, tartarate resistant acid phosphatase, hydroxyproline) and urine (calcium). The bone loss was accompanied by reduced serum estradiol levels and bone TGF-β3 content. Preventive and curative treatment with raloxifene ameliorated bony alterations and was more effective than CVD. Deprived estrogen levels (that in turn reduced lumbar TGF-β3 content) following PHT and SVP, thus, might represent one of the various mechanisms of AEDs-induced bone loss. Raloxifene preserved the bony changes without interfering with their antiepileptic efficacy, and hence raloxifene could be a potential therapeutic option in the management of PHT and SVP-induced bone disease if clinically approved.Keywords: antiepileptic drugs, osteoporosis, raloxifene, TGF-β3
Procedia PDF Downloads 344797 Design and Development of Novel Anion Selective Chemosensors Derived from Vitamin B6 Cofactors
Authors: Darshna Sharma, Suban K. Sahoo
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The detection of intracellular fluoride in human cancer cell HeLa was achieved by chemosensors derived from vitamin B6 cofactors using fluorescence imaging technique. These sensors were first synthesized by condensation of pyridoxal/pyridoxal phosphate with 2-amino(thio)phenol. The anion recognition ability was explored by experimental (UV-VIS, fluorescence and 1H NMR) and theoretical DFT [(B3LYP/6-31G(d,p)] methods in DMSO and mixed DMSO-H2O system. All the developed sensors showed both naked-eye detectable color change and remarkable fluorescence enhancement in the presence of F- and AcO-. The anion recognition was occurred through the formation of hydrogen bonded complexes between these anions and sensor, followed by the partial deprotonation of sensor. The detection limit of these sensors were down to micro(nano) molar level of F- and AcO-.Keywords: chemosensors, fluoride, acetate, turn-on, live cells imaging, DFT
Procedia PDF Downloads 400796 Effect of Sodium Alginate-based Edible Coating with Natural Essential Oils and Modified Atmosphere Packaging on Quality of Fresh-cut Pineapple
Authors: Muhammad Rafi Ullah Khan, Yaodong Guo, Vanee Chonhenchob, Jinjin Pei, Chongxing Huang
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The effect of sodium alginate (1%) based edible coating incorporated natural essential oils; thymol, carvone and carvacrol as antimicrobial agents at different concentrations (0.1, 0.5 and 1.0 %) on the quality changes of fresh-cut pineapple were investigated. Pineapple dipped in distilled water was served as control. After coating, fruit were sealed in a modified atmosphere package (MAP) using high permeable film; and stored at 5 °C. Gas composition in package headspace, color values (L*, a*, b*, C*), TSS, pH, ethanol, browning, and microbial decay were monitored during storage. Oxygen concentration continuously decreased while carbon dioxide concentration inside all packages continuously increased over time. Color parameters (L*, b*, c*) decreased and a* values increased during storage. All essential oils significantly (p ≤ 0.05) prevented microbial growth than control. A significantly higher (p ≤ 0.05) ethanol content was found in the control than in all other treatments. Visible microbial growth, high ethanol, and low color values limited the shelf life to 6 days in control as compared to 9 days in all other treatments. Among all essential oils, thymol at all concentrations maintained the overall quality of the pineapple and could potentially be used commercially in fresh fruit industries for longer storage.Keywords: essential oils, antibrowning agents, antimicrobial agents, modified atmosphere packaging, microbial decay, pineapple
Procedia PDF Downloads 59795 Effects of Adding Sodium Nitroprusside in Semen Diluents on Motility, Viability and Lipid Peroxidation of Sperm of Holstein Bulls
Authors: Leila Karshenas, Hamid Reza Khodaei, Behnaz Mahdavi
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We know that nitric oxide (NO) plays an important role in all sexual activities of animals. It is made in body from NO synthase enzyme and L-arginin molecule. NO can bound with sulfur-iron complexes and because production of steroid sexual hormones is related to enzymes which have this complex, NO can change the activity of these enzymes. NO affects many cells including endothelial cells of veins, macrophages and mast cells. These cells are found in testis leydig cells and therefore are important source of NO in testis tissue. Minimizing damages to sperm at the time of sperm freezing and thawing is really important. The goal of this study was to determine the function of NO before freezing and its effects on quality and viability of sperms after thawing and incubation. 4 Holstein bulls were selected from the age of 4, and artificial insemination was done for 3 weeks (2 times a week). Treatments were 0, 10, 50 and 100 nm of sodium nitroprusside (SNP). Data analysis was performed by SAS98 program. Also, mean comparison was done using Duncan's multiple ranges test (P<0.05). Concentrations used was found to increase motility and viability of spermatozoa at 1, 2 and 3 hours after thawing significantly (P<0.05), but there was no significant difference at zero time. SNP levels reduced the amount of lipid peroxidation in sperm membrane, increased acrosome health and improved sample membranes especially in 50 and 100 nm treatments. According to results, adding SNP to semen diluents increases motility and viability of spermatozoa. Also, it reduces lipid peroxidation in sperm membrane and improves sperm function.Keywords: sperm motility, nitric oxide, lipid peroxidation, spermatozoa
Procedia PDF Downloads 359794 Catalytic Synthesis and Characterization of N-(4-(Tert-Butyl) Benzyl)-1-(4-Tert-Butyl) Phenyl)-N-Methyl Methanaminium Chloride from Tert-Butyl Benzyl Derivatives
Authors: Muhammad A. Muhammad
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Butenafine (N-4-tert-butyl benzyl-N-methyl-1-naphthylene methylamine hydrochloride) is a benzylamine antimycotic (antifungal) agent that has a broad spectrum of action. The quest for improved antimycotic action brought about many research on the structure-activity properties of butenafine in relation to other antifungal agents. Of all those research, only little or no effort was recorded on the substituents attached to the aromatic systems in butenafine. In this research, N-(4-(tert-butyl) benzyl)-1-(4-tert-butyl) phenyl)-N-methyl methanaminium chloride, which is a butenafine analogue was synthesised from tert-butyl benzyl derivatives, by reductive amination using various solvents through a direct approach, where 1,2-dichloroethane gave the best solvent action at 40 °C (Yield: 75%) and of all the reducing agents used, sodium borohydride was found to give the best reducing action in the presence of silica chloride at room temperature (Yield: 50%). Characterization of the compound by 1H NMR showed a singlet peak of 18 hydrogen atoms with a chemical shift at 1.3-1.5 ppm for the presence of 6 methyl groups in the two tert-butyl substituents, the 13C NMR also indicated the presence of the two tert-butyl substituents by the peak with a chemical shift at 31-32 ppm for the six methyl carbon atoms, the IR indicated the presence of a tertiary ammonium ion by a strong band at 2460 cm-1 and finally the EIS-MS confirmed the molar mass of the compound by a mass to charge ratio of 324.2693. These results suggested that the target molecule was actually synthesised and therefore, 1,2-dichloroethane is a good solvent for this synthesis, and the most suitable reducing agent is sodium borohydride.Keywords: antimicrobial agents, antimycotic agents, butenafine, chemotherapeutic agents, semisynthetic agents
Procedia PDF Downloads 292793 Surface Sterilization Retain Postharvest Quality and Shelf Life of Strawberry and Cherry Tomato during Modified Atmosphere Packaging
Authors: Ju Young Kim, Mohammad Zahirul Islam, Mahmuda Akter Mele, Su Jeong Han, Hyuk Sung Yoon, In-Lee Choi, Ho-Min Kang
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Strawberry and tomato fruits were harvested at the red ripens maturity stage in the Republic of Korea. The fruits were dipped in fungi solution and afterwards were sterilized with sodium hypochlorite (NaOCl) and chlorine dioxide (ClO2) gas. Some fruits were dipped in 150μL/L NaOCl solution for 10 minutes, and others were treated with 5μL/L ClO2 gas for 12 hours and packed with 20,000 cc OTR (oxygen transmission rate) film, the rest were packed in 10,000 cc OTR film inserted with 5μL/L ClO2 gas. 5μL/L ClO2 gas insert treatment showed the lowest carbon dioxide and ethylene, and the highest oxygen concentration was on the final storage day (15th day) in both strawberry and tomato fruits. Tomato fruits showed the lowest fresh weight loss in 5μL/L ClO2 gas insert treatment. The visual quality as well as shelf life showed the highest in 5μL/L ClO2 gas insert treatment of both strawberry and tomato fruits. In addition, the fungal incidence of strawberry and tomato fruits were the most suppressed in 5μL/L ClO2 gas insert treatment. 5μL/L ClO2 gas insert treatment showed higher firmness and soluble solids in both strawberry and tomato fruits. So, 5μL/L ClO2 gas insert treatment may be useful to prevent the fungal incidence as well as retaining the postharvest quality, and increase the shelf life of strawberry and tomato fruits for long term storage. This study was supported by Export Promotion Technology Development Program (314027-03), IPET, Ministry of Agriculture, Food and Rural Affairs, Republic of Korea.Keywords: chlorine dioxide, ethylene, fungi, sodium hypochlorite
Procedia PDF Downloads 366792 Comparing the Effectiveness of the Crushing and Grinding Route of Comminution to That of the Mine to Mill Route in Terms of the Percentage of Middlings Present in Processed Lead-Zinc Ore Samples
Authors: Chinedu F. Anochie
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The presence of gangue particles in recovered metal concentrates has been a serious challenge to ore dressing engineers. Middlings lower the quality of concentrates, and in most cases, drastically affect the smelter terms, owing to exorbitant amounts paid by Mineral Processing industries as treatment charge. Models which encourage optimization of liberation operations have been utilized in most ore beneficiation industries to reduce the presence of locked particles in valuable concentrates. Moreover, methods such as incorporation of regrind mills, scavenger, rougher and cleaner cells, to the milling and flotation plants has been widely employed to tackle these concerns, and to optimize the grade–recovery relationship of metal concentrates. This work compared the crushing and grinding method of liberation, to the mine to mill route, by evaluating the proportion of middlings present in selectively processed complex Pb-Zn ore samples. To establish the effect of size reduction operations on the percentage of locked particles present in recovered concentrates, two similar samples of complex Pb- Zn ores were processed. Following blasting operation, the first ore sample was ground directly in a ball mill (Mine to Mill Route of Comminution), while the other sample was manually crushed, and subsequently ground in the ball mill (Crushing and Grinding Route of Comminution). The two samples were separately sieved in a mesh to obtain the desired representative particle sizes. An equal amount of each sample that would be processed in the flotation circuit was then obtained with the aid of a weighing balance. These weighed fine particles were simultaneously processed in the flotation circuit using the selective flotation technique. Sodium cyanide, Methyl isobutyl carbinol, Sodium ethyl xanthate, Copper sulphate, Sodium hydroxide, Lime and Isopropyl xanthate, were the reagents used to effect differential flotation of the two ore samples. Analysis and calculations showed that the degree of liberation obtained for the ore sample which went through the conventional crushing and grinding route of comminution, was higher than that of the directly milled run off mine (ROM) ore. Similarly, the proportion of middlings obtained from the separated galena (PbS) and sphalerite (ZnS) concentrates, were lower for the crushed and ground ore sample. A concise data which proved that the mine to mill method of size reduction is not the most ideal technique for the recovery of quality metal concentrates has been established.Keywords: comminution, degree of liberation, middlings, mine to mill
Procedia PDF Downloads 133791 Evaluation of Groundwater and Seawater Intrusion at Tajoura Area, NW, Libya
Authors: Abdalraheem Huwaysh, Khalil Al Samarrai, Yasmin ElAhmar
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Water quality is an important factor that determines its usage for domestic, agricultural and industrial uses. This study was carried out through the Tajoura Area, Jifarah Plain, Northwest Libya. Chemical and physical parameters were measured and analyzed for groundwater samples collected in 2021 from twenty-six wells distributed throughout the investigation area. Overexploitation of groundwater caused considerable deterioration in the water quality, especially at Tajoura Town (20 Km east of Tripoli). The aquifer shows an increase in salinization, which has reached an alarming level in many places during the past 25 years as a result of the seawater intrusion. The chemical composition of the water samples was compared with the drinking water standards of WHO and Libyan Standards. Groundwater from this area was not suitable to be a source for direct drinking based on Total Dissolved Solids. The dominant cation is sodium, while the dominant anion is chloride. Based on the Piper trilinear diagram, most of the groundwater samples (90%) were identified as sodium chloride type. The best groundwater quality exists at the southern part of the study area. Serious degradation in the water quality, expressed in salinity increase, occurs as we go towards the coastline. The abundance of NaCl waters is strong evidence to attribute the successive deterioration of the water quality to the seawater intrusion. Considering the values of Cl- concentration and the ratio of Cl-/HCO3-, about 70% of the groundwater samples were strongly affected by the saline water. Car wash stations in the study area as well as the unlined disposal pond used for the collection of untreated wastewater, contribute significantly to the deterioration of water quality. The water quality in this area needs to be monitored regularly and it is crucial to treat the water before consumption.Keywords: Tajoura, groundwater, seawater intrusion, water quality
Procedia PDF Downloads 104790 Study on Technological Development for Reducing the Sulfur Dioxide Residue Problem in Fresh Longan for Exporting
Authors: Wittaya Apai, Satippong Rattanakam, Suttinee Likhittragulrung, Nuttanai Tungmunkongvorakul, Sompetch Jaroensuk
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The objective of this study was to find some alternative ways to decrease sulfur dioxide (SO₂) residue problem and prolong storage life in fresh longan for export. Office of Agricultural Research and Development Region 1, Chiang Mai province conducted the research and development from 2016-2018. A grade longan cv. Daw fruit with panicle attached was placed in 11.5 kg commercial perforated plastic basket. They had 5 selected treatments comprising of 3 baskets as replication for each treatment, i.e. 1.5% SO₂ fumigation prior to insert SO₂-generated pads (Uvasys®) (1.5% SO₂+SO₂ pad), dipping in 5% hydrochloric acid (HCl) mixed with 1% sodium metabisulfite (SMS) for 5 min (5% HCl +1% SMS), ozone (O₃) fumigation for 1 hours (h) prior to 1.5% SO₂ fumigation (O₃ 1 h+1.5% SO₂), 1.5% SO₂ fumigation prior to O₃ fumigation for 1 h (1.5% SO₂+O₃ 1 h) and 1.5% SO₂ fumigation alone as commercial treatment (1.5% SO₂). They were stored at 5 ˚C, 90% relative humidity (RH) for 40-80 days. The results found that the possible treatments were 1.5% SO₂+O₃ 1 h and 5% HCl +1% SMS respectively and prevented pericarp browning for 80 days at 5 ºC. There were no significant changes in some parameters in any treatments; 1.5% SO₂+O₃ 1 h and 1.5% SO₂ during storage, i.e., pericarp browning, flesh discoloration, disease incidence (%) and sensory evaluation during storage. Application 1.5% SO₂+O₃ 1 h had a tendency less both SO₂ residue in fruit and disease incidence (%) including brighter pericarp color as compared with commercial 1.5% SO₂ alone. Moreover, HCl 5%+SMS 1% showed the least SO₂ residue in whole fruit below codex tolerance at 50 mg/kg throughout period of time. The fruit treated with 1.5% SO₂+O₃ 1 h, 1.5% SO₂, 5% HCl+1% SMS, O₃ 1 h+1.5% SO₂, and 1.5% SO₂+SO₂ pad could prolong storage life for 40, 40, 40, 30 and 30 days respectively at 5°C, 90% RH. Thus, application 1.5% SO₂+O₃ 1 h and/or 5% HCl +1% SMS could be used for extending shelf life fresh longan exported to restricted countries due to less SO₂ residue and fruit quality was maintained as compared with the conventional method.Keywords: longan, sulfur dioxide, ozone fumigation, sodium metabisulfite
Procedia PDF Downloads 126789 Human Beta Defensin 1 as Potential Antimycobacterial Agent against Active and Dormant Tubercle Bacilli
Authors: Richa Sharma, Uma Nahar, Sadhna Sharma, Indu Verma
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Counteracting the deadly pathogen Mycobacterium tuberculosis (M. tb) effectively is still a global challenge. Scrutinizing alternative weapons like antimicrobial peptides to strengthen existing tuberculosis artillery is urgently required. Considering the antimycobacterial potential of Human Beta Defensin 1 (HBD-1) along with isoniazid, the present study was designed to explore the ability of HBD-1 to act against active and dormant M. tb. HBD-1 was screened in silico using antimicrobial peptide prediction servers to identify its short antimicrobial motif. The activity of both HBD-1 and its selected motif (Pep B) was determined at different concentrations against actively growing M. tb in vitro and ex vivo in monocyte derived macrophages (MDMs). Log phase M. tb was grown along with HBD-1 and Pep B for 7 days. M. tb infected MDMs were treated with HBD-1 and Pep B for 72 hours. Thereafter, colony forming unit (CFU) enumeration was performed to determine activity of both peptides against actively growing in vitro and intracellular M. tb. The dormant M. tb models were prepared by following two approaches and treated with different concentrations of HBD-1 and Pep B. Firstly, 20-22 days old M. tbH37Rv was grown in potassium deficient Sauton media for 35 days. The presence of dormant bacilli was confirmed by Nile red staining. Dormant bacilli were further treated with rifampicin, isoniazid, HBD-1 and its motif for 7 days. The effect of both peptides on latent bacilli was assessed by colony forming units (CFU) and most probable number (MPN) enumeration. Secondly, human PBMC granuloma model was prepared by infecting PBMCs seeded on collagen matrix with M. tb(MOI 0.1) for 10 days. Histopathology was done to confirm granuloma formation. The granuloma thus formed was incubated for 72 hours with rifampicin, HBD-1 and Pep B individually. Difference in bacillary load was determined by CFU enumeration. The minimum inhibitory concentrations of HBD-1 and Pep B restricting growth of mycobacteria in vitro were 2μg/ml and 20μg/ml respectively. The intracellular mycobacterial load was reduced significantly by HBD-1 and Pep B at 1μg/ml and 5μg/ml respectively. Nile red positive bacterial population, high MPN/ low CFU count and tolerance to isoniazid, confirmed the formation of potassium deficienybaseddormancy model. HBD-1 (8μg/ml) showed 96% and 99% killing and Pep B (40μg/ml) lowered dormant bacillary load by 68.89% and 92.49% based on CFU and MPN enumeration respectively. Further, H&E stained aggregates of macrophages and lymphocytes, acid fast bacilli surrounded by cellular aggregates and rifampicin resistance, indicated the formation of human granuloma dormancy model. HBD-1 (8μg/ml) led to 81.3% reduction in CFU whereas its motif Pep B (40μg/ml) showed only 54.66% decrease in bacterial load inside granuloma. Thus, the present study indicated that HBD-1 and its motif are effective antimicrobial players against both actively growing and dormant M. tb. They should be further explored to tap their potential to design a powerful weapon for combating tuberculosis.Keywords: antimicrobial peptides, dormant, human beta defensin 1, tuberculosis
Procedia PDF Downloads 263788 Pomegranate Peel Based Edible Coating Treatment for Safety and Quality of Chicken Nuggets
Authors: Muhammad Sajid Arshad, Sadaf Bashir
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In this study, the effects of pomegranate peel based edible coating were determined on safety and quality of chicken nuggets. Four treatment groups were prepared as control (without coating), coating with sodium alginate (SA) (1.5%), pomegranate peel powder (PPP) (1.5%), and combination of SA and PPP. There was a significant variation observed with respect to coating treatments and storage intervals. The chicken nuggets were subjected to refrigerated storage (40C) and were analyzed at regular intervals of 0, 7, 14 1 and 21 days. The microbiological quality was determined by total aerobic and coliform counts. Total aerobic (5.09±0.05 log CFU/g) and coliforms (3.91±0.06 log CFU/g) counts were higher in uncoated chicken nuggets whereas lower was observed in coated chicken nuggets having combination of SA and PPP. Likewise, antioxidants potential of chicken nuggets was observed by assessing total phenolic contents (TPC) and DPPH activity. Higher TPC (135.66 GAE/100g) and DPPH (64.65%) were found in combination with SA and PPP, whereas minimum TPC (91.38) and DPPH (41.48) was observed in uncoated chicken nuggets. Regarding the stability analysis of chicken nuggets, thiobarbituric acid reactive substances (TBARS) and peroxide value (POV) were determined. Higher TBARS (1.62±0.03 MDA/Kg) and POV (0.92±0.03 meq peroxide/kg) were found in uncoated chicken nuggets. Hunter color values were also observed in both uncoated and coated chicken nuggets. Sensorial attributes were also observed by the trained panelists. The higher sensory score for appearance, color, taste, texture and overall acceptability were observed in control (uncoated) while in coated treatments, it was found within acceptable limits. In nutshell, the combination of SA and PPP enhanced the overall quality, antioxidant potential, and stability of chicken nuggets.Keywords: chicken nuggets, edible coatings, pomegranate peel powder, sodium alginate
Procedia PDF Downloads 148787 Mixed Monolayer and PEG Linker Approaches to Creating Multifunctional Gold Nanoparticles
Authors: D. Dixon, J. Nicol, J. A. Coulter, E. Harrison
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The ease with which they can be functionalized, combined with their excellent biocompatibility, make gold nanoparticles (AuNPs) ideal candidates for various applications in nanomedicine. Indeed several promising treatments are currently undergoing human clinical trials (CYT-6091 and Auroshell). A successful nanoparticle treatment must first evade the immune system, then accumulate within the target tissue, before enter the diseased cells and delivering the payload. In order to create a clinically relevant drug delivery system, contrast agent or radiosensitizer, it is generally necessary to functionalize the AuNP surface with multiple groups; e.g. Polyethylene Glycol (PEG) for enhanced stability, targeting groups such as antibodies, peptides for enhanced internalization, and therapeutic agents. Creating and characterizing the biological response of such complex systems remains a challenge. The two commonly used methods to attach multiple groups to the surface of AuNPs are the creation of a mixed monolayer, or by binding groups to the AuNP surface using a bi-functional PEG linker. While some excellent in-vitro and animal results have been reported for both approaches further work is necessary to directly compare the two methods. In this study AuNPs capped with both PEG and a Receptor Mediated Endocytosis (RME) peptide were prepared using both mixed monolayer and PEG linker approaches. The PEG linker used was SH-PEG-SGA which has a thiol at one end for AuNP attachment, and an NHS ester at the other to bind to the peptide. The work builds upon previous studies carried out at the University of Ulster which have investigated AuNP synthesis, the influence of PEG on stability in a range of media and investigated intracellular payload release. 18-19nm citrate capped AuNPs were prepared using the Turkevich method via the sodium citrate reduction of boiling 0.01wt% Chloroauric acid. To produce PEG capped AuNPs, the required amount of PEG-SH (5000Mw) or SH-PEG-SGA (3000Mw Jenkem Technologies) was added, and the solution stirred overnight at room temperature. The RME (sequence: CKKKKKKSEDEYPYVPN, Biomatik) co-functionalised samples were prepared by adding the required amount of peptide to the PEG capped samples and stirring overnight. The appropriate amounts of PEG-SH and RME peptide were added to the AuNP to produce a mixed monolayer consisting of approximately 50% PEG and 50% RME. The PEG linker samples were first fully capped with bi-functional PEG before being capped with RME peptide. An increase in diameter from 18-19mm for the ‘as synthesized’ AuNPs to 40-42nm after PEG capping was observed via DLS. The presence of PEG and RME peptide on both the mixed monolayer and PEG linker co-functionalized samples was confirmed by both FTIR and TGA. Bi-functional PEG linkers allow the entire AuNP surface to be capped with PEG, enabling in-vitro stability to be achieved using a lower molecular weight PEG. The approach also allows the entire outer surface to be coated with peptide or other biologically active groups, whilst also offering the promise of enhanced biological availability. The effect of mixed monolayer versus PEG linker attachment on both stability and non-specific protein corona interactions was also studied.Keywords: nanomedicine, gold nanoparticles, PEG, biocompatibility
Procedia PDF Downloads 339786 The Hydrotrope-Mediated, Low-Temperature, Aqueous Dissolution of Maize Starch
Authors: Jeroen Vinkx, Jan A. Delcour, Bart Goderis
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Complete aqueous dissolution of starch is notoriously difficult. A high-temperature autoclaving process is necessary, followed by cooling the solution below its boiling point. The cooled solution is inherently unstable over time. Gelation and retrogradation processes, along with aggregation-induced by undissolved starch remnants, result in starch precipitation. We recently observed the spontaneous gelatinization of native maize starch (MS) in aqueous sodium salicylate (NaSal) solutions at room temperature. A hydrotropic mode of solubilization is hypothesized. Differential scanning calorimetry (DSC) and polarized optical microscopy (POM) of starch dispersions in NaSal solution were used to demonstrate the room temperature gelatinization of MS at different concentrations of MS and NaSal. The DSC gelatinization peak shifts to lower temperatures, and the gelatinization enthalpy decreases with increasing NaSal concentration. POM images confirm the same trend through the disappearance of the ‘Maltese cross’ interference pattern of starch granules. The minimal NaSal concentration to induce complete room temperature dissolution of MS was found to be around 15-20 wt%. The MS content of the dispersion has little influence on the amount of NaSal needed to dissolve it. The effect of the NaSal solution on the MS molecular weight was checked with HPSEC. It is speculated that, because of its amphiphilic character, NaSal enhances the solubility of MS in water by association with the more hydrophobic MS moieties, much like urea, which has also been used to enhance starch dissolution in alkaline aqueous media. As such small molecules do not tend to form micelles in water, they are called hydrotropes rather than surfactants. A minimal hydrotrope concentration (MHC) is necessary for the hydrotropes to structure themselves in water, resulting in a higher solubility of MS. This is the case for the system MS/NaSal/H₂O. Further investigations into the putative hydrotropic dissolution mechanism are necessary.Keywords: hydrotrope, dissolution, maize starch, sodium salicylate, gelatinization
Procedia PDF Downloads 187785 Dietary N-6/N-3 PUFA Ratios Affect the Homeostasis of CD4+ T Cells in Mice with Dextran Sulfate Sodium-Induced Colitis
Authors: Cyoung-Huei Huang, Chiu-Li Yeh, Man-Hui Pai, Sung-Ling Yeh
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This study evaluated the effect of different dietary n-6/n-3 polyunsaturated fatty acid (PUFA) ratios on modulating helper T (Th) and regulatory T (Treg) lymphocytes in mice with dextran sulfate sodium (DSS)-induced colitis. There were 3 control and 3 colitis groups in this study. Mice were fed for 24 d with an AIN-93G diet either with soybean oil (S), a mixture of soybean oil and low fish oil content (LF) or high fish oil content (HF). The ratio of n-6/n-3 PUFA in the LF diet was 4:1, and that in the HF diet was 2:1. The control groups drank distilled water while colitis groups provided 2% DSS in drinking water during day 15-19. All mice drank distilled water from day 20-24 for recovery and sacrificed on day 25. The results showed that colitis resulted in higher Th1, Th2, and Th17 and lower Treg percentages in the blood. Also, plasma haptoglobin and proinflammatory chemokines were elevated in colon lavage fluid. Colitic groups with fish oil had lower inflammatory mediators in the plasma and colon lavage fluid. Further, the percentages of Th1, Th2, and Th17 cells in the blood were lower, whereas Treg cell percentages were higher than those in the soybean oil group. The colitis group with n-6/n-3 PUFA ratio 2:1 had more pronounce effects than ratio 4:1. These results suggest that diets with an n-6/n-3 PUFA ratio of 2:1 or 4:1 regulate the Th/Treg balance and attenuate inflammatory mediator production in colitis. Compared to the n-6/n-3 PUFA ratio 4:1, the ratio of 2:1 was more effective in reducing inflammatory reactions in DSS-induced colitis.Keywords: inflammatory bowel disease, n-3 polyunsaturated fatty acids, helper T lymphocyte, regulatory T lymphocyte
Procedia PDF Downloads 297784 Mannose-Functionalized Lipopolysaccharide Nanoparticles for Macrophage-Targeted Dual Delivery of Rifampicin and Isoniazid
Authors: Mumuni Sumaila, Viness Pillay, Yahya E. Choonara, Pradeep Kumar, Pierre P. Kondiah
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Tuberculosis (TB) remains a serious challenge to public health globally, despite every effort put together to curb the disease. Current TB therapeutics available have proven to be inefficient due to a multitude of drawbacks that range from serious adverse effects/drug toxicity to inconsistent bioavailability, which ultimately contributes to the emergence of drug-resistant TB. An effective ‘cargo’ system designed to cleverly deliver therapeutic doses of anti-TB drugs to infection sites and in a sustained-release manner may provide a better therapeutic choice towards winning the war against TB. In the current study, we investigated mannose-functionalized lipopolysaccharide hybrid nanoparticles for safety and efficacy towards macrophage-targeted simultaneous delivery of the two first-line anti-TB drugs, rifampicin (RF) and isoniazid (IS). RF-IS-loaded lipopolysaccharide hybrid nanoparticles were fabricated using the solvent injection technique (SIT), incorporating soy lecithin (SL) and low molecular weight chitosan (CS) as the lipid and polysaccharide components, respectively. Surface-functionalized nanoparticles were obtained through the reaction of the aldehyde group of mannose with free amine functionality present at the surface of the nanoparticles. The functionalized nanocarriers were spherical with average particle size and surface charge of 107.83 nm and +21.77 mV, respectively, and entrapment efficiencies (EE) were 53.52% and 69.80% for RF and IS, respectively. FTIR spectrum revealed high-intensity bands between 1663 cm⁻¹ and 1408 cm⁻¹ wavenumbers (absent in non-functionalized nanoparticles), which could be attributed to the C=N stretching vibration produced by the formation of Schiff’s base (–N=CH–) during the mannosylation reaction. In vitro release studies showed a sustained-release profile for RF and IS, with less than half of the total payload released over a 48-hour period. The nanocarriers were biocompatible and safe, with more than 80% cell viability achieved when incubated with RAW 264.7 cells at concentrations 30 to 500 μg/mL over a 24-hour period. Cellular uptake studies (after a 24-hour incubation period with the murine macrophage cells, RAW 264.7) revealed a 13- and a 9-fold increase in intracellular accumulation of RF and IS, respectively, when compared with the unformulated RF+IS solution. A 6- and a 3-fold increase in intracellular accumulation of RF and IS, respectively, were observed when compared with the non-functionalized nanoparticles. Furthermore, fluorescent microscopy images showed nanoparticle internalization and accumulation within the RAW 264.7 cells, which was more significant in the mannose-functionalized system compared to the non-functionalized nanoparticles. The overall results suggested that the fabricated mannose-functionalized lipopolysaccharide nanoparticles are a safe and promising platform for macrophage-targeted delivery of anti-TB therapeutics. However, in vivo pharmacokinetic/pharmacodynamics studies are required to further substantiate the therapeutic efficacy of the nanosystem.Keywords: anti-tuberculosis therapeutics, hybrid nanosystem, lipopolysaccharide nanoparticles, macrophage-targeted delivery
Procedia PDF Downloads 172783 Effect of Coated Sodium Butyrate (CM3000®) On Zootechnical Performance, Immune Status and Necrotic Enteritis After Experimental Infection of Broiler Chickens
Authors: Mohamed Ahmed Tony, Mohamed Hamoud
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The present study was conducted to determine the effect of commercially coated slow-release sodium butyrate (CM3000®) as a feed additive on zootechnical performance, immune status and Clostridium perfringens severity after experimental infection. Three hundred 1-d-old broiler chicks (Cobb 500) were randomly distributed into 3 treatment groups (4 replicates each) using 25 chicks per replicate on floor pens. Control (C) birds were offered non-supplemented basal diets. Treatments 1 and 2 (T1 and T2) were fed diets containing CM3000® at 300 and 500 g/ton feed, respectively, during the entire experimental period (35 days). Feed and water were offered ad-libitum. Feed consumption and body weight were recorded weekly to calculate body weight gain and feed conversion. Blood samples were collected to evaluate the immune status of the birds against Newcastle disease vaccines using HI test. At the end of the experimental period, 20 birds were chosen randomly from each group (5 birds from each pen) to compare carcass yield. At day 16 of age 20 birds from each group (5 birds/replicate) were bacteriologically examined and proved to be free from Clostridium perfringens. The isolated birds were challenged orally with 1 ml buffer containing 106 CFU/ml Clostridium perfringens local isolate and prepared from necrotic enteritis (NE) diseased farms. Birds were observed on a regular basis daily for any signs of NE. Birds that died in the challenged group were necropsied to determine the cause of death. On day 28 of age, the surviving chickens were killed by cervical dislocation and necropsied immediately. Intestinal tracts were removed and intestinal lesions were scored. Tissue samples of the duodenum, jejunum, ileum and cecum for histopathological examination were collected. All collected data were statistically analyzed using IBM SPSS® version 19 software for personal computers. Means were compared by one-way ANOVA (P<0.05) followed by the Duncan Post Hoc test. The results revealed that body weight gain was significantly (P<0.05) improved in chicks fed on both doses of CM3000® compared to the control one. Final body weight gain in T1 and T2 were 2064.94 and 2141.37 g/bird, respectively, while in the control group, the weight gain showed 1952.78 g/bird. In addition, supplementation of diets with CM3000® increased significantly feed intake (P<0.05). Total feed intake in T1 and T2 were 3186.32 and 3273.29 g/bird, respectively; however, feed intake in the control group recorded 3081.95 g/bird. The best feed conversion was recorded in T2 group (1.53). Feed conversion in the control and T1 groups were 1.58 and 1.54, respectively. Dressing percentage, liver weights and the other carcasses yields were not different between treatments. The butyrate significantly enhanced immune responses measured against Newcastle disease vaccines. Sodium butyrate significantly reduced NE lesions and healthy improved the intestinal tissues in the samples collected from T1 and T2-challenged chickens versus those collected from the control group. In conclusion, exogenous administration of slow-release butyrate (CM3000®) is capable of improving performance, enhancing immunity and NE disease resistance in broiler chickens.Keywords: sodium butyrate, broiler chicken, zootechnical performance, immunity, necrotic enteritis
Procedia PDF Downloads 83782 Studies of Carbohydrate, Antioxidant, Nutrient and Genomic DNA Characterization of Fresh Olive Treated with Alkaline and Acidic Solvent: An Innovation
Authors: A. B. M. S. Hossain, A. Abdelgadir, N. A. Ibrahim
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Fresh ripen olive cannot be consumed immediately after harvest due to the excessive bitterness having polyphenol as antioxidant. Industrial processing needs to be edible the fruit. The laboratory processing technique has been used to make it edible by using acid (vinegar, 5% acetic acid) and alkaline solvent (NaOH). Based on the treatment and consequence, innovative data have been found in this regard. The experiment was conducted to investigate biochemical content, nutritional and DNA characterization of olive fruit treated with alkaline (Sodium chloride anhydrous) and acidic solvent (5% acetic acid, vinegar). The treatments were used as control (no water), water control, 10% sodium chloride anhydrous (NaOH), vinegar (5% acetic acid), vinegar + NaOH and vinegar + NaOH + hot water treatment. Our results showed that inverted sugar and glucose content were higher in the vinegar and NaOH treated olive than in other treatments. Fructose content was the highest in vinegar + NaOH treated fruit. Nutrient contents NO3 K, Ca and Na were found higher in the treated fruit than the control fruit. Moreover, maximum K content was observed in the case of all treatments compared to the other nutrient content. The highest acidic (lower pH) condition (sour) was found in treated fruit. DNA yield was found higher in water control than acid and alkaline treated olives. DNA band was wider in the olive treated water control compared to the NaOH, vinegar, vinegar + NaOH and vinegar + NaOH + Hot water treatment. Finally, results suggest that vinegar + NaOH treated olive fruit was the best for fresh olive homemade processing after harvesting for edible purpose.Keywords: olive, vinegar, sugars, DNA band, bioprocess biotechnology
Procedia PDF Downloads 185781 Continuum-Based Modelling Approaches for Cell Mechanics
Authors: Yogesh D. Bansod, Jiri Bursa
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The quantitative study of cell mechanics is of paramount interest since it regulates the behavior of the living cells in response to the myriad of extracellular and intracellular mechanical stimuli. The novel experimental techniques together with robust computational approaches have given rise to new theories and models, which describe cell mechanics as a combination of biomechanical and biochemical processes. This review paper encapsulates the existing continuum-based computational approaches that have been developed for interpreting the mechanical responses of living cells under different loading and boundary conditions. The salient features and drawbacks of each model are discussed from both structural and biological points of view. This discussion can contribute to the development of even more precise and realistic computational models of cell mechanics based on continuum approaches or on their combination with microstructural approaches, which in turn may provide a better understanding of mechanotransduction in living cells.Keywords: cell mechanics, computational models, continuum approach, mechanical models
Procedia PDF Downloads 363780 Mechanisms of O-1602 Induced Endothelium-Independent Vasorelaxation of Rat Small Mesenteric Artery
Authors: Yousuf Al Suleimani, Ahmed Al Mahruqi
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A typical cannabinoid O-1602 induces vasorelaxation and activates the orphan G protein-coupled receptor GPR55 in human endothelial cells. The aim of this study is to characterize the mechanisms of endothelium-independent relaxation of O-1602 in the rat small mesenteric artery using wire myograph. In endothelium-denuded vessels, O-1602 partially produced concentration-dependent vasorelaxation. In vessels depleted of intracellular Ca2+ (by EGTA and methoxamine), CaCl2 produced concentration-dependent contraction. Preincubation with O-1602 (at 10 µM and 30 µM) abolished the contractile responses (P<0.01). The putative antagonist at novel “endothelial anandamide receptor” O-1918 (10 µM) significantly reversed the inhibitory effect of O-1602 on CaCl2-induced vasoconstriction. It is likely that the mechanism of endothelium-independent vasorelaxation to O-1602 is mediated by interfering with Ca2+ entry via an O-1918-sensitive pathway.Keywords: O-1602, endothelium, vasorelaxation, calcium
Procedia PDF Downloads 359779 The Effect of the Flow Pipe Diameter on the Rheological Behavior of a Polymeric Solution (CMC)
Authors: H. Abchiche, M. Mellal
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The aim of this work is to study the parameters that influence the rheological behavior of a complex fluid (sodium Carboxyméthylcellulose solution), on a capillary rheometer. An installation has been made to be able to vary the diameter of trial conducts. The obtained results allowed us to deduce that: the diameter of trial conducts have a remarkable effect on the rheological responds.Keywords: bingham’s fluid, CMC, cylindrical conduit, rheological behavior
Procedia PDF Downloads 332778 Effect of Sodium Hydroxide Treatment on the Mechanical Properties of Crushed and Uncrushed Luffa cylindrica Fibre Reinforced rLDPE Composites
Authors: Paschal A. Ubi, Salawu Abdul Rahman Asipita
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The use of suitable engineering materials which poses less harm to ,an and the environment is sort for in recent times, thus giving rise to polymer composites filled with natural organic reinforcement which are biodegradable. Treatment of natural fibres is essential in improving matrix to filler adhesion, hence improving its mechanical properties. In this study, investigations were carried out to determine the effect of sodium hydroxide treatment on the tensile, flexural, impact and hardness properties of crushed and uncrushed luffa cylindrica fibre reinforced recycled low density polyethylene composites. The LC (Luffa Cylindrica) fibres were treated with 0%, 2%, 4%, 6%, 8%, and 10% wt. NaOH concentrations for a period of 24 hours under room temperature conditions. The compounding of the waste LDPE was done using a two roll mill at a temperature of 150 oC and cured in a hydraulic press at a temperature of 150oC for 3 minutes at 3 metric tonnes. A formulation of 20/80g (reinforcement to matrix ratio in grams) was maintained for all fabricated samples. Analysis of the results showed that the uncrushed luffa fibre samples gave better mechanical properties compared with the crushed luffa fibre samples. The uncrushed luffa fibre composites had optimum tensile and flexural strengths of 7.65MPa and 17.08Mpa respectively corresponding to a young modulus and flexural modulus of 21.08MPa and 232.22MPa for the 8% and 4%wt. NaOH concentration respectively. Results obtained in the research showed that NaOH treatment with the 8% NaOH concentration improves the mechanical properties of the LC fibre reinforced composites when compared with other NaOH treatment concentration values.Keywords: LC fibres, NaOH concentration, LC/rLDPE composite, tensile strength, flexural strength
Procedia PDF Downloads 281777 Measurements of Scattering Cross Sections for 5.895 keV Photons in Various Polymers
Authors: H. Duggal, G. Singh, G. Singh, A. Bhalla, S. Kumar, J. S. Shahi, D. Mehta
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The total differential cross section for scattering of the 5.895 keV photons by various polymers has been measured at scattering angle of 135o. The experimental measurements were carried out using the energy dispersive setup involving annular source of the 55Fe radioisotope and a low energy germanium (LEGe) detector. The cross section values are measured for 20 polymer targets namely, Paraffin Wax, Polytetrafluoro ethylene (PTFE), Cellulose, Silicone oil, Polyvinyl alcohol (PVA), Polyvinyl purrolidone (PVP), Polymethyl methacrylate (PMMA), Kapton, Mylar, Chitosan, Polyvinyl chloride (PVC), Bakelite, Carbopol, Chlorobutyl rubber (CBR), Polyetylene glycol (PEG), Polysorbate-20, Nylon-6, Cetyl alcohol, Carboxyl methyl sodium cellulose and Sodium starch glucolate. The measurements were performed in vacuum so as to avoid scattering contribution due to air and strong absorption of low energy photons in the air column. In the present investigations, the geometrical factor and efficiency of the detector were determined by measuring the K x-rays emitted from the 22Ti and 23V targets excited by the Mn K x-rays in the same experimental set up. The measured scattering cross sections have been compared with the sum of theoretically calculated elastic and inelastic scattering cross sections. The theoretical elastic (Rayleigh) scattering cross sections based on the various form factor approximations, namely, non-relativistic form factor (NF), relativistic form factor (RF), modified form factor (MF), and MF with anomalous scattering factor (ASF) as well as the second order S-matrix formalisms, and the inelastic scattering differential cross sections based on the Klein-Nishina formula after including the inelastic scattering function (KN+ISF) have been calculated. The experimental results show fairly good agreement with theoretical cross sections.Keywords: photon, polymers, elastic and inelastic, scattering cross sections
Procedia PDF Downloads 689776 Potentiometric Determination of Moxifloxacin in Some Pharmaceutical Formulation Using PVC Membrane Sensors
Authors: M. M. Hefnawy, A. M. A. Homoda, M. A. Abounassif, A. M. Alanazia, A. Al-Majed, Gamal A. E. Mostafa
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PVC membrane sensors using different approach e.g. ion-pair, ionophore, and Schiff-base has been used as testing membrane sensor. Analytical applications of membrane sensors for direct measurement of variety of different ions in complex biological and environmental sample are reported. The most important step of such PVC membrane sensor is the sensing active material. The potentiometric sensors have some outstanding advantages including simple design, operation, wide linear dynamic range, relative fast response time, and rotational selectivity. The analytical applications of these techniques to pharmaceutical compounds in dosage forms are also discussed. The construction and electrochemical response characteristics of Poly (vinyl chloride) membrane sensors for moxifloxacin HCl (MOX) are described. The sensing membranes incorporate ion association complexes of moxifloxacin cation and sodium tetraphenyl borate (NaTPB) (sensor 1), phosphomolybdic acid (PMA) (sensor 2) or phosphotungstic acid (PTA) (sensor 3) as electroactive materials. The sensors display a fast, stable and near-Nernstian response over a relative wide moxifloxacin concentration range (1 ×10-2-4.0×10-6, 1 × 10-2-5.0×10-6, 1 × 10-2-5.0×10-6 M), with detection limits of 3×10-6, 4×10-6 and 4.0×10-6 M for sensor 1, 2 and 3, respectively over a pH range of 6.0-9.0. The sensors show good discrimination of moxifloxacin from several inorganic and organic compounds. The direct determination of 400 µg/ml of moxifloxacin show an average recovery of 98.5, 99.1 and 98.6 % and a mean relative standard deviation of 1.8, 1.6 and 1.8% for sensors 1, 2, and 3 respectively. The proposed sensors have been applied for direct determination of moxifloxacin in some pharmaceutical preparations. The results obtained by determination of moxifloxacin in tablets using the proposed sensors are comparable favorably with those obtained using the US Pharmacopeia method. The sensors have been used as indicator electrodes for potentiometric titration of moxifloxacin.Keywords: potentiometry, PVC, membrane sensors, ion-pair, ionophore, schiff-base, moxifloxacin HCl, sodium tetraphenyl borate, phosphomolybdic acid, phosphotungstic acid
Procedia PDF Downloads 439775 Mathematical Modelling of the Effect of Glucose on Pancreatic Alpha-Cell Activity
Authors: Karen K. Perez-Ramirez, Genevieve Dupont, Virginia Gonzalez-Velez
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Pancreatic alpha-cells participate on glucose regulation together with beta cells. They release glucagon hormone when glucose level is low to stimulate gluconeogenesis from the liver. As other excitable cells, alpha cells generate Ca2+ and metabolic oscillations when they are stimulated. It is known that the glucose level can trigger or silence this activity although it is not clear how this occurs in normal and diabetic people. In this work, we propose an electric-metabolic mathematical model implemented in Matlab to study the effect of different glucose levels on the electrical response and Ca2+ oscillations of an alpha cell. Our results show that Ca2+ oscillations appear in opposite phase with metabolic oscillations in a window of glucose values. The model also predicts a direct relationship between the level of glucose and the intracellular adenine nucleotides showing a self-regulating pathway for the alpha cell.Keywords: Ca2+ oscillations, mathematical model, metabolic oscillations, pancreatic alpha cell
Procedia PDF Downloads 178774 Cellular Targeting to Dual Gaseous Microenvironments by Polydimethylsiloxane Microchip
Authors: Samineh Barmaki, Ville Jokinen, Esko Kankuri
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We report a microfluidic chip that can be used to modify the gaseous microenvironment of a cell-culture in ambient atmospheric conditions. The aim of the study is to show the cellular response to nitric oxide (NO) under hypoxic (oxygen < 5%) condition. Simultaneously targeting to hypoxic and nitric oxide will provide an opportunity for NO‑based therapeutics. Studies on cellular responses to lowered oxygen concentration or to gaseous mediators are usually carried out under a specific macro environment, such as hypoxia chambers, or with specific NO donor molecules that may have additional toxic effects. In our study, the chip consists of a microfluidic layer and a cell culture well, separated by a thin gas permeable polydimethylsiloxane (PDMS) membrane. The main design goal is to separate the gas oxygen scavenger and NO donor solutions, which are often toxic, from the cell media. Two different types of gas exchangers, titled 'pool' and 'meander' were tested. We find that the pool design allows us to reach a higher level of oxygen depletion than meander (24.32 ± 19.82 %vs -3.21 ± 8.81). Our microchip design can make the cells culture more simple and makes it easy to adapt existing cell culture protocols. Our first application is utilizing the chip to create hypoxic conditions on targeted areas of cell culture. In this study, oxygen scavenger sodium sulfite generates hypoxia and its effect on human embryonic kidney cells (HEK-293). The PDMS membrane was coated with fibronectin before initiating cell cultures, and the cells were grown for 48h on the chips before initiating the gas control experiments. The hypoxia experiments were performed by pumping of O₂-depleted H₂O into the microfluidic channel with a flow-rate of 0.5 ml/h. Image-iT® reagent as an oxygen level responser was mixed with HEK-293 cells. The fluorescent signal appears on cells stained with Image-iT® hypoxia reagent (after 6h of pumping oxygen-depleted H₂O through the microfluidic channel in pool area). The exposure to different levels of O₂ can be controlled by varying the thickness of the PDMS membrane. Recently, we improved the design of the microfluidic chip, which can control the microenvironment of two different gases at the same time. The hypoxic response was also improved from the new design of microchip. The cells were grown on the thin PDMS membrane for 30 hours, and with a flowrate of 0.1 ml/h; the oxygen scavenger was pumped into the microfluidic channel. We also show that by pumping sodium nitroprusside (SNP) as a nitric oxide donor activated under light and can generate nitric oxide on top of PDMS membrane. We are aiming to show cellular microenvironment response of HEK-293 cells to both nitric oxide (by pumping SNP) and hypoxia (by pumping oxygen scavenger solution) in separated channels in one microfluidic chip.Keywords: hypoxia, nitric oxide, microenvironment, microfluidic chip, sodium nitroprusside, SNP
Procedia PDF Downloads 133773 Effect of Chemical Mutagen on Seeds Germination of Lima Bean
Authors: G. Ultanbekova, Zh. Suleimenova, Zh. Rakhmetova, G. Mombekova, S. Mantieva
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Plant Growth Promoting Rhizobacteria (PGPR) are a group of free-living bacteria that colonize the rhizosphere, enhance plant growth of many cereals and other important agricultural crops and protect plants from disease and abiotic stresses through a wide variety of mechanisms. The use of PGPR has been proven to be an environmentally sound way of increasing crop yields by facilitating plant growth. In the present study, strain improvement of PGPR isolates were carried out by chemical mutagenesis for the improvement of growth and yield of lima bean. Induced mutagenesis is widely used for the selection of microorganisms producing biologically active substances and further improving their activities. Strain improvement is usually done by classical mutagenesis which involves exposing the microbes to chemical or physical mutagens. The strains of Pseudomonas putida 4/1, Azotobacter chroococcum Р-29 and Bacillus subtilis were subjected to mutation process for strain improvement by treatment with a chemical agent (sodium nitrite) to cause mutation and were observed for its consequent action on the seeds germination and plant growth of lima bean (Phaseolus lunatus). Bacterial mutant strains of Pseudomonas putida M-1, Azotobacter chroococcum M-1 and Bacillus subtilis M-1, treated with sodium nitrite in the concentration of 5 mg/ml for 120 min, were found effective to enhance the germination of lima bean seeds compared to parent strains. Moreover, treatment of the lima bean seeds with a mutant strain of Bacillus subtilis M-1 had a significant stimulation effect on plant growth. The length of the stems and roots of lima bean treated with Bacillus subtilis M-1 increased significantly in comparison with parent strain in 1.6 and 1.3 times, respectively.Keywords: chemical mutagenesis, germination, kidney bean, plant growth promoting rhizobacteria (PGPR)
Procedia PDF Downloads 198772 Modeling of Glycine Transporters in Mammalian Using the Probability Approach
Authors: K. S. Zaytsev, Y. R. Nartsissov
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Glycine is one of the key inhibitory neurotransmitters in Central nervous system (CNS) meanwhile glycinergic transmission is highly dependable on its appropriate reuptake from synaptic cleft. Glycine transporters (GlyT) of types 1 and 2 are the enzymes providing glycine transport back to neuronal and glial cells along with Na⁺ and Cl⁻ co-transport. The distribution and stoichiometry of GlyT1 and GlyT2 differ in details, and GlyT2 is more interesting for the research as it reuptakes glycine to neuron cells, whereas GlyT1 is located in glial cells. In the process of GlyT2 activity, the translocation of the amino acid is accompanied with binding of both one chloride and three sodium ions consequently (two sodium ions for GlyT1). In the present study, we developed a computer simulator of GlyT2 and GlyT1 activity based on known experimental data for quantitative estimation of membrane glycine transport. The trait of a single protein functioning was described using the probability approach where each enzyme state was considered separately. Created scheme of transporter functioning realized as a consequence of elemental steps allowed to take into account each event of substrate association and dissociation. Computer experiments using up-to-date kinetic parameters allowed receiving the number of translocated glycine molecules, Na⁺ and Cl⁻ ions per time period. Flexibility of developed software makes it possible to evaluate glycine reuptake pattern in time under different internal characteristics of enzyme conformational transitions. We investigated the behavior of the system in a wide range of equilibrium constant (from 0.2 to 100), which is not determined experimentally. The significant influence of equilibrium constant in the range from 0.2 to 10 on the glycine transfer process is shown. The environmental conditions such as ion and glycine concentrations are decisive if the values of the constant are outside the specified range.Keywords: glycine, inhibitory neurotransmitters, probability approach, single protein functioning
Procedia PDF Downloads 119771 Promotion of Lipid Syntheses of Microalgae by Microfluidic-Assisted Membrane Distortion
Authors: Seul Ki Min, Gwang Heum Yoon, Jung Hyun Joo, Hwa Sung Shin
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Cellular membrane distortion is known as a factor to change intracellular signaling. However, progress of relevant studies is difficult because there are no facilities that can control membrane distortion finely. In this study, we developed microfluidic device which can inflict mechanical stress on cell membrane of Chlamydomonas reinhardtii using regular height of the channels. And cellular physiological changes were analyzed from cells cultured in the device. Excessive calcium ion influx through into cytoplasm was induced from mechanical stress. The results revealed that compressed cells had up-regulated Mat3 mRNA which regulates cell size and cell cycle from a prolonged G1 phase. Additionally, TAG used for the production of biodiesel was raised rapidly from 4 h after compression. Taken together, membrane distortion can be considered as an attractive inducer for biofuel production.Keywords: mechanical stress, membrane distortion, Chlamydomonas reinhardtii, deflagellation, cell cycle, lipid metabolism
Procedia PDF Downloads 375770 Partitioning of Non-Metallic Nutrients in Lactating Crossbred Cattle Fed Buffers
Authors: Awadhesh Kishore
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The goal of the study was to determine how different non-metallic nutrients are partitioned from feed in various physiological contexts and how buffer addition in ruminant nutrition affects these processes. Six lactating crossbred dairy cows were selected and divided into three groups on the basis of their phenotypic and productive features (374±14 kg LW). Two treatments, T1 and T2, were randomly assigned to one animal from each group. Animals under T1 and T2 were moved to T2 and T1, respectively, after 30 days. T2 was the only group to receive buffers containing magnesium oxide and sodium bicarbonate at 0.0 and 0.01% of LW (the real amounts are equivalent to 75.3±4.0 and 30 7.7±2.0 g/d, respectively). T1 was used as the control. Wheat straw and berseem were part of the base diet, whereas wheat grain and mustard cake were part of the concentrate mixture. Following a 21-day feeding period, metabolic and milk production trials were carried out for seven consecutive days. The Kearl equation used the urine's calorific value to determine its volume. Chemical analyses were performed to determine the levels of nitrogen, carbohydrates, calories, and phosphorus in samples of feed, waste, buffer, mineral mixture, water, feces, urine, and milk that were collected. The information was analyzed statistically. Notable results included decreased nitrogen and carbohydrate partitioning to feces from feed, while increased calorie partitioning to milk and body storage, and increased carbohydrate partitioning to body storage. Phosphorus balance was significantly better in T2. The application of buffers in ruminant diets was found to increase the output of calories in milk, as well as the number of calories and carbohydrates stored in the body, while decreasing the amount of nitrogen in faeces. As a result, it may be advised to introduce buffers to feed crossbred dairy cattle.Keywords: cattle, Magnesium oxide, non-metallic nutrients, partitioning, Sodium bicarbonate
Procedia PDF Downloads 58