Search results for: total lipid

Authors: Hawraa Zbeeb, Hala Khalifeh, Mohamad Khalil, Francesca Storace, Francesca Baldini, Giulio Lupidi, Laura Vergani

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Sarcopoterium spinosum, a widely distributed spiny shrub belonging to the Rosaceae family, is rich in essential and beneficial constituents. In fact, S. spinosum fruits and roots are traditionally used as herbal medicine in the eastern Mediterranean landscape, and this shrub is mentioned as a medicinal plant in a large number of ethnobotanical surveys. Aqueous root extracts from S. spinosum are used by traditional medicinal practitioners for weight loss treatment of diabetes and pain. Moreover, the anti-diabetic activity of S. spinosum root extract has been reported in different studies, but the beneficial effects of aerial parts, especially fruits, have not been elucidated yet. The aim of the present study was to investigate the in vitro antioxidant and lipid-lowering properties of an ethanolic extract from S. spinosum fruits using both hepatic (FaO) and endothelial (HECV) cells in an attempt to evaluate its possible employment as a nutraceutical supplement. First of all, in vitro spectrophotometric assays were employed to characterize the extract. The total phenol content (TPC) was evaluated by Folin–Ciocalteu spectrophotometric method and the radical scavenging activity was tested by 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2, 2'-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) assays. After that, the beneficial effects of the extract were tested on cells. FaO cells treated for 3 hours with 0.75 mM oleate/palmitate mix (1:2 molar ratio) mimic in vitro a moderate hepato-steatosis. HECV cells exposed for 1 hour to 100 µM H₂O₂ mimic an oxidative insult leading to oxidative stress conditions. After the metabolic and oxidative insult, both cell lines were treated with increasing concentrations of the S. spinosum extract (1, 10, 25 µg/mL) for 24 hours. The results showed the S. spinosum ethanolic extract is rather rich in phenols (TPC of 18.6 mgGAE/g dry extracts). Moreover, the extract showed a good scavenging ability in vitro (IC₅₀ 15.9 µg/ml and 10.9 µg/ml measured by DPPH and ABTS assays, respectively). When the extract was tested on cells, the results showed that it could ameliorate some markers of cell dysfunction. The three concentrations of the extract led to a significant decrease in the intracellular triglyceride (TG) content in steatotic FaO cells measured by spectrophotometric assay. On the other hand, HECV cells treated with increasing concentrations of the extract did not result in a significant decrease in both lipid peroxidation measured by the Thiobarbituric Acid Reactive Substances (TBARS) assay, and in reactive oxygen species (ROS) production measured by fluorometric analysis after DCF staining. Interestingly, the ethanolic extract was able to accelerate the wound repair of confluent HECV cells with respect to H₂O₂-insulted cells as measured by T-scratch assay. Taken together, these results seem to indicate that the ethanol extract from S. spinosum fruits is rich in phenol compounds and plays considerable lipid-lowering activity in vitro on steatotic hepatocytes and accelerates wound healing repair on endothelial cells. In light of that, the ethanolic extract from S. spinosum fruits could be a potential candidate for nutraceutical applications.

Keywords: antioxidant activity, ethanolic extract, lipid-lowering activity, phenolic compounds, Sarcopoterium spinosum fruits

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Authors: H. Jeries, N. Volkova, C. G. Iglesias, M. Najjar, M. Rosenblat, M. Aviram, T. Hayek

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Background: Synthetic forms of glucocorticoids (e.g., prednisone, prednisolone) are anti-inflammatory drugs which are widely used in clinical practice. The role of glucocorticoids (GCs) in cardiovascular diseases including atherosclerosis is highly controversial, and their impact on macrophage foam cell formation is still unknown. Our aim was to investigate the effects of prednisone or its active metabolite, prednisolone, on macrophage oxidative stress and lipid metabolism using in-vivo, ex-vivo and in-vitro systems. Methods: The in-vivo study included C57BL/6 mice which were intraperitoneally injected with prednisone or prednisolone (5mg/kg) for 4 weeks, followed by lipid metabolism analyses in the mice aorta, and in peritoneal macrophages (MPM). In the ex-vivo study, we analyzed the effect of serum samples obtained from 9 healthy volunteers before or after treatment with oral prednisone (20mg for 5 days), on J774A.1 macrophage atherogenicity. In-vitro studies were conducted using J774A.1 macrophages, human monocyte derived macrophages (HMDM) and fibroblasts. Cells were incubated with increasing concentrations (0-200 ng/ml) of prednisone or prednisolone, followed by determination of cellular oxidative status, triglyceride and cholesterol metabolism. Results: Prednisone or prednisolone treatment resulted in a significant reduction in triglycerides and mainly in cholesterol cellular accumulation in MPM or in J774A.1 macrophages incubated with human serum. Similar resulted were noted in HMDM or in J774A.1 macrophages which were directly incubated with the GCs. These effects were associated with GCs inhibitory effect on triglycerides and cholesterol biosynthesis rates, throughout downregulation of diacylglycerol acyltransferase1 (DGAT1) expression, and of the sterol regulatory element binding protein (SREBP2) and HMGCR expression, respectively. In parallel to prednisone or prednisolone induced reduction in macrophage triglyceride content, paraoxonase 2 (PON2) expression was significantly upregulated. GCs-induced reduction of cellular triglyceride and cholesterol mass was mediated by the GCs receptors on macrophages since the GCs receptor antagonist (RU 486) abolished these effects. In fibroblasts, unlike macrophages, prednisone or prednisolone showed no anti-atherogenic effects. Conclusions: Prednisone or prednisolone are anti-atherogenic since they protected macrophages from lipid accumulation and foam cell formation.

Keywords: atherosclerosis, cholesterol, foam cell, macrophage, prednisone, prednisolone, triglycerides

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Authors: Samar Saadeldin Abdelmotalab Omer, Ikram Mohammed Eltayeb Elsiddig, Amna Beshir Medani Ahmed, Saad Mohammed Hussein Ayoub

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Many herbal medicinal products are considered as potential hypocholesterolemic agents with encouraging safety profiles, however, only a limited amount of clinical research exists to support their efficacy. The present study was designed to compare the antihypercholesterolaemic and antioxidant activities of the crude ethanolic extracts of Citrus reticulata peel, Zingiber officinale rhizome, and Sesamum indicum seeds. Forty-five rats were used throughout the experiment, which were divided into nine groups, five rats in each as follows; normal control group (normal rats fed with standard normal diet), rats fed hypercholesterolemic diet consisting of 1% cholesterol and 10% saturated animal fat, which were further divided into eight groups; hypercholesterolemic control group (rats only fed hypercholesterolemic diet), groups 3,4,5,6,7, and 8 were given Citrus reticulata, Zingiber officinale, and Sesamum indicum ethanolic extracts at doses of (250mg/kg and 500mg/kg, respectively) orally; and group 9 rats were given atorvastatin (0.18mg/kg) orally as a reference antihypercholesterolaemic drug. Blood samples were obtained four weeks following treatment from the retro-orbital venous plexus after fasting overnight from all groups and the lipid profile (serum total cholesterol (TC), high-density-lipoprotein-cholesterol (HDL-C), low-density lipoprotein-cholesterol (LDL-C), and triglycerides levels) was measured and the risk ratio (TC/HDL-C) was assessed. The antioxidant activity of the three plant extracts was determined using DPPH free-radical assay. Results of in vivo and in vitro antihypercholesterolaemic and antioxidant assay, respectively, revealed that the three extracts possess comparable antioxidant and anti-hypercholesterolaemic activities.

Keywords: anti hypercholesterolemic effects, antioxidant activity, HDL, LDL, TC, TGs, citrus reticulata, sesamum indicum, zingiber officinale

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Authors: Pinar Ercan, Sedef N. El

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The goals of this study were to determine the total anthocyanin and procyanidin contents and their in vitro bioaccessibilities of apple, red grape and cinnamon by a static in vitro digestion method reported by the COST FA1005 Action INFOGEST, as well as in vitro inhibitory effects of these food samples on starch and lipid digestive enzymes. While the highest total anthocyanin content was found in red grape (164.76 ± 2.51 mg/100 g), the highest procyanidin content was found in cinnamon (6432.54±177.31 mg/100 g) among the selected food samples (p<0.05). The anthocyanin bioaccessibilities were found as 10.23±1 %, 8.23±0.64 %, and 8.73±0.70 % in apple, red grape, and cinnamon, respectively. The procyanidin bioaccessibilities of apple, red grape, and cinnamon were found as 17.57±0.71 %, 14.08±0.74 % and 18.75±1.49 %, respectively. The analyzed apple, red grape and cinnamon showed the inhibitory activity against α-glucosidase (IC50 544.27±21.94, 445.63±15.67, 1592±17.58 μg/mL, respectively), α-amylase (IC50 38.41±7.26, 56.12±3.60, 3.54±0.86 μg/mL, respectively), and lipase (IC50 52.65±2.05, 581.70±54.14, 49.63±2.72 μg/mL, respectively). Red grape sample showed the highest inhibitory activity against α-glucosidase, cinnamon showed the highest inhibitory activity against α-amylase and lipase according to IC50 (concentration of inhibitor required to produce a 50% inhibition of the initial rate of reaction) and Catechin equivalent inhibition capacity (CEIC50) values. This study reported that apple, grape and cinnamon samples can inhibit the activity of digestive enzymes in vitro. The consumption of these samples would be used in conjunction with a low-calorie diet for body weight management.

Keywords: anthocyanin, α-amylase, α-glucosidase, lipase, procyanidin

Procedia PDF Downloads 148

Authors: Faheema Khan

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We have investigated the effects of salt stress on the stability of plant growth, water relations, photosynthetic variables, lipid peroxidation and antioxidant system in salt-tolerant (PK-327) and salt-sensitive (PK-471) soybean genotypes. Ten-day-old salt-tolerant and salt-sensitive soybean plants were subjected to 0-150 mM NaCl for 15 days. While the growth of genotype PK-327 was not affected significantly up to 75 mM NaCl treatment, the growth of the PK-471 was reduced significantly beyond 25 mM NaCl treatments. Salt stress caused severe impairments in photosynthetic variables like photosynthetic rate, chlorophyll fluorescence and chlorophyll content, being more pronounced in salt-sensitive genotype than in salt-tolerant.The activities of antioxidant enzymes (superoxide dismutase, catalase, ascorbate peroxidase and glutathione reductase) were higher in PK-327 than in PK-471 at various levels of salt treatments.It is concluded that tolerance capacity of PK-327 against salinity can be associated with the ability of this genotype in keeping an active photosynthetic system and strong antioxidant defence system.

Keywords: salt stress, soybean, antioxidant, photosynthesis

Procedia PDF Downloads 342

Authors: Talita Kato, Moises Grespan, Elza I. Ida, Massami Shimokomaki, Adriana L. Soares

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White Striping (WS) is becoming a concern for the poultry industry, as it affects the look of breast broiler chicken meat leading it to rejection by the consumers. It is characterized by the appearance of varying degrees of white striations on the Pectoralis major muscle surface following the direction of the muscle fiber. The etiology of this myopathy is still unknown, however it is suggested to be associated with increased weight gain rate and age of the bird, attributing the phenomenon to the genetically bird’s selection for efficiently higher meat production. The aim of this study was to evaluate the occurrence of Pectoralis major WS in a commercial plant in southern Brazil and its chemical characterization. The breast meat samples (n=660) from birds of 47 days of age, were classified as: Normal NG (no apparent white striations), Moderate MG (when the fillets present thin lines <1 mm) and Severe SG (white striations present ˃1 mm thick covering a large part of the fillet surface). Thirty samples (n = 10 for each level of severity) were analyzed for pH, color (L*, a*, b*), proximate chemical composition (moisture, protein, ash and lipids contents) and hydroxyproline in order to determine the collagen content. The results revealed the occurrence for NG group was 16.97%, 51.67% for MG group and 31.36% for SG group. Although the total protein content did not differ significantly, the collagen index was 42% higher in favor to SG in relation to NG. Also the lipid fraction was 27% higher for SG group. The NG presented the lowest values of the parameters L* and a* (P ≤ 0.05), as there was no white striations on its surface and highest b* value in SG, because of the maximum lipid contents. These results indicate there was a contribution of the SG muscle cells to oversynthesize connective tissue components on the muscle fascia. In conclusion, this study revealed a high incidence of White Striping on broiler commercial line in Brazil thus, there is a need to identify the causes of this abnormality in order to diminish or to eliminate it.

Keywords: collagen content, commercial line, pectoralis major muscle, proximate composition

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Authors: Carolina N. França, Jônatas B. do Amaral, Maria C.O. Izar, Ighor L. Teixeira, Francisco A. Fonseca

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Introduction: Atherosclerosis is a progressive disease, characterized by lipid and fibrotic element deposition in large-caliber arteries. Conditions related to the development of atherosclerosis, as dyslipidemia, hypertension, diabetes, and smoking are associated with endothelial dysfunction. There is a frequent recurrence of cardiovascular outcomes after acute myocardial infarction and, at this sense, cycles of mobilization of monocyte subtypes (classical, intermediate and nonclassical) secondary to myocardial infarction may determine the colonization of atherosclerotic plaques in different stages of the development, contributing to early recurrence of ischemic events. The recruitment of different monocyte subsets during inflammatory process requires the expression of chemokine receptors CCR2, CCR5, and CX3CR1, to promote the migration of monocytes to the inflammatory site. The aim of this study was to evaluate the effect of lipid-lowering treatment by six months in the monocyte phenotype and chemokine receptor levels of patients after Acute Myocardial Infarction (AMI). Methods: This is a PROBE (prospective, randomized, open-label trial with blinded endpoints) study (ClinicalTrials.gov Identifier: NCT02428374). Adult patients (n=147) of both genders, ageing 18-75 years, were randomized in a 2x2 factorial design for treatment with rosuvastatin 20 mg/day or simvastatin 40 mg/day plus ezetimibe 10 mg/day as well as ticagrelor 90 mg 2x/day and clopidogrel 75 mg, in addition to conventional AMI therapy. Blood samples were collected at baseline, after one month and six months of treatment. Monocyte subtypes (classical - inflammatory, intermediate - phagocytic and nonclassical – anti-inflammatory) were identified, quantified and characterized by flow cytometry, as well as the expressions of the chemokine receptors (CCR2, CCR5 and CX3CR1) were also evaluated in the mononuclear cells. Results: After six months of treatment, there was an increase in the percentage of classical monocytes and reduction in the nonclassical monocytes (p=0.038 and p < 0.0001 Friedman Test), without differences for intermediate monocytes. Besides, classical monocytes had higher expressions of CCR5 and CX3CR1 after treatment, without differences related to CCR2 (p < 0.0001 for CCR5 and CX3CR1; p=0.175 for CCR2). Intermediate monocytes had higher expressions of CCR5 and CX3CR1 and lower expression of CCR2 (p = 0.003; p < 0.0001 and p = 0.011, respectively). Nonclassical monocytes had lower expressions of CCR2 and CCR5, without differences for CX3CR1 (p < 0.0001; p = 0.009 and p = 0.138, respectively). There were no differences after the comparison between the four treatment arms. Conclusion: The data suggest a time-dependent modulation of classical and nonclassical monocytes and chemokine receptor levels. The higher percentage of classical monocytes (inflammatory cells) suggest a residual inflammatory risk, even under preconized treatments to AMI. Indeed, these changes do not seem to be affected by choice of the lipid-lowering strategy.

Keywords: acute myocardial infarction, chemokine receptors, lipid-lowering treatment, monocyte subtypes

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Authors: Zahra Azizi, Ashkan Karbasi, Farzin Firouzian, Sara Soleimani Asl, Akram Ranjbar

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Background: One of the most often used herbicides in agriculture is paraquat (PQ), which is very harmful to both people and animals. Chitosan is a well-known, non-toxic polymer commonly used in preparing particles via ionotropic gelation facilitated by negatively charged agents such as sodium alginate. This study aimed to compare the effects of PQ and nanoparaquat (PQNPs) on liver function in male rats. Materials & Methods: Rats were exposed to PQ & PQNPs (4 mg/kg/day, intraperitoneally) for seven days. Then, rats were anesthetized, and serum and liver samples were collected. Later, enzymatic activities such as alanine transaminase (ALT), aspartate transaminase (AST), and alkaline phosphatase (ALP) in serum and oxidative stress biomarkers such as lipid peroxidation (LPO), total antioxidant capacity (TAC) and total thiol groups (TTG) levels in liver tissue were measured by colorimetric methods. Also, histological changes in the liver were evaluated. Results: PQ altered the levels of ALT, AST, and ALP while inducing oxidative stress in the liver. Additionally, liver homogenates with PQ exposure had challenged LPO, TAC, and TTG levels. The severe liver damage is indicated by a significant increase in the enzyme activity of AST, ALT, and ALP in serum. According to the results of the current study, PQNPs, as compared to PQ and the control group, lowered ALT, AST, ALP, and LPO levels while increasing TAC and TTG levels. Conclusion: According to biochemical and histological investigations, PQ loaded in chitosan-alginate particles is more efficient than free PQ at reducing liver toxicity.

Keywords: paraquat, paraquat nanoparticles, liver, oxidative stress

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Authors: Linda A. O’Higgins, Astrid Wingler, Jorge Oliveira

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The growth of Chlorella vulgaris was characterized as a function of irradiance in a laboratory turbidostat (1 L) and compared to batch growth in sunlit modules (5–25 L) of the commercial Phytobag photobioreactor. The effects of variable sunlight and culture density were deconvoluted by a mathematical model. The analysis showed that algal growth was light-limited due to shading by external construction elements and due to light attenuation within the algal bags. The model was also used to predict maximum biomass productivity. The manipulative experiments and the model predictions were confronted with data from a production season of a 10m2 pilot-scale photobioreactor, Phytobag (10,000 L). The analysis confirmed light limitation in all three photobioreactors. An additional limitation of biomass productivity was caused by the nitrogen starvation that was used to induce lipid accumulation. Reduction of shading and separation of biomass and lipid production are proposed for future optimization.

Keywords: microalgae, batch cultivation, Chlorella vulgaris, Mathematical model, photobioreactor, scale-up

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Authors: Seema Dhariwal, Kiran Maan, Ruchi Baghel, Apoorva Sharma, Poonam Rana

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Introduction: Traumatic brain injury (TBI) is defined as the temporary or permanent alteration in brain function and pathology caused by an external mechanical force. It represents the leading cause of mortality and morbidity among children and youth individuals. Various models of TBI in rodents have been developed in the laboratory to mimic the scenario of injury. Blast overpressure injury is common among civilians and military personnel, followed by accidents or explosive devices. In addition to this, the lateral Controlled cortical impact (CCI) model mimics the blunt, penetrating injury. Method: In the present study, we have developed two different mild TBI models using blast and CCI injury. In the blast model, helium gas was used to create an overpressure of 130 kPa (±5) via a shock tube, and CCI injury was induced with an impact depth of 1.5mm to create diffusive and focal injury, respectively. C57BL/6J male mice (10-12 weeks) were divided into three groups: (1) control, (2) Blast treated, (3) CCI treated, and were exposed to different injury models. Serum was collected on Day1 and day7, followed by biphasic extraction using MTBE/Methanol/Water. Prepared samples were separated on Charged Surface Hybrid (CSH) C18 column and acquired on UHPLC-Q-TOF-MS using ESI probe with inhouse optimized parameters and method. MS peak list was generated using Markerview TM. Data were normalized, Pareto-scaled, and log-transformed, followed by multivariate and univariate analysis in metaboanalyst. Result and discussion: Untargeted profiling of lipids generated extensive data features, which were annotated through LIPID MAPS® based on their m/z and were further confirmed based on their fragment pattern by LipidBlast. There is the final annotation of 269 features in the positive and 182 features in the negative mode of ionization. PCA and PLS-DA score plots showed clear segregation of injury groups to controls. Among various lipids in mild blast and CCI, five lipids (Glycerophospholipids {PC 30:2, PE O-33:3, PG 28:3;O3 and PS 36:1 } and fatty acyl { FA 21:3;O2}) were significantly altered in both injury groups at Day 1 and Day 7, and also had VIP score >1. Pathway analysis by Biopan has also shown hampered synthesis of Glycerolipids and Glycerophospholipiods, which coincides with earlier reports. It could be a direct result of alteration in the Acetylcholine signaling pathway in response to TBI. Understanding the role of a specific class of lipid metabolism, regulation and transport could be beneficial to TBI research since it could provide new targets and determine the best therapeutic intervention. This study demonstrates the potential lipid biomarkers which can be used for injury severity diagnosis and identification irrespective of injury type (diffusive or focal).

Keywords: LipidBlast, lipidomic biomarker, LIPID MAPS®, TBI

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Authors: A. El-Gendy, S. Al-Farraj, S. Al Kahtani, M. El-Hedeny

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Contaminations of persistent organic pollutants (POPs) such as, dichlorodiphenyl trichloroethane (DDT), hexachlorocyclohexane (HCH) and chlordane (CHLs) were examined in the edible mantle tissues of the commercial cuttlefish Sepia pharaonis Ehrenberg 1831, collected from the marine water of the Arabian Gulf. The mean concentrations of DDT, CHLs and HCH were in the ranges of 29.4 - 56 ng/g, 47.4 - 100 ng/g and 1 - 4 ng/g, respectively. Among the POPs analyzed, HCH showed the lowest concentrations ranging between 1 to 5 ng/g lipid wt. However, concentrations of DDT, CHLs and HCH, detected in this study, were generally comparable or lower than those found in studies of similar cephalopod species from other areas subject to a high anthropogenic impact. Relationships between total body lengths and/or dorsal mantle lengths of the organisms and the concentration values of the studied POPs were also considered. Compared with recommendations of the international organizations, there are no potential risks associated with consuming the studied cuttlefish species.

Keywords: cuttlefish, Sepia pharaonis, organochlorine, DDT, CHLs, HCH, Arabian Gulf

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Authors: Rattanamanee Chomchan, Sunisa Siripongvutikorn, Panupong Puttarak

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Ricegrass or young rice sprouts can be introduced as one of functional product since cereal sprouts have been much interested in this era due to their high nutritive values. Bio-fortification of selenium is one strategy to improve plant bioactive compounds. However, the level of selenium used are varied among species of plants, hence, the proper level need to be investigated. In this current study, influence of selenium bio-fortification hydroponically in the form of sodium selenite following the range 0, 10, 20, 30 and 40 mg Se/L on growth characteristics, selenium content, total extractable phenolic content (TPC) accumulation, lipid peroxidation and anti-oxidative properties of ricegrass were investigated. Results revealed that selenium bio-fortified exogenously increased the accumulation of selenium in ricegrass by 5.3 fold at 40 mg Se/L treatment without significant changes in leaves biomass at harvesting day while root part weight were slightly decreased when increased selenium level, respectively. Selenium at low concentration (10 and 20 mg Se/L) can stimulate the production of phenolic compounds and antioxidant activities in young ricegrass as measured by DPPH, ABTS and FRAP assay. Conversely, higher level of selenium fortification reduced the accumulation of phenolics in ricegrass afterward by acting as pro-oxidant. Moreover, highest significant reduction in oxidative stress, measured as malondialdehyde content was also observed at 20 mg Se/L treatment which in correlation to high TPC and antioxidant activities. In conclusion, selenium bio-fortification can be used as a technique to improve precious to ricegrass.

Keywords: antioxidant activities, bio-fortification, ricegrass, selenium

Procedia PDF Downloads 248

Authors: Jeanne Leblond, Warren Viricel, Amira Mbarek

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Although several products have reached the market, gene therapeutics are still in their first stages and require optimization. It is possible to improve their lacking efficiency by the use of carefully engineered vectors, able to carry the genetic material through each of the biological barriers they need to cross. In particular, getting inside the cell is a major challenge, because these hydrophilic nucleic acids have to cross the lipid-rich plasmatic and/or endosomal membrane, before being degraded into lysosomes. It takes less than one hour for newly endocytosed liposomes to reach highly acidic lysosomes, meaning that the degradation of the carried gene occurs rapidly, thus limiting the transfection efficiency. We propose to use a new pH-sensitive lipid able to change its conformation upon protonation at endosomal pH values, leading to the disruption of the lipidic bilayer and thus to the fast release of the nucleic acids into the cytosol. It is expected that this new pH-sensitive mechanism promote endosomal escape of the gene, thereby its transfection efficiency. The main challenge of this work was to design a preparation presenting fast-responding lipidic bilayer destabilization properties at endosomal pH 5 while remaining stable at blood pH value and during storage. A series of pH-sensitive lipids able to perform a conformational switch upon acidification were designed and synthesized. Liposomes containing these switchable lipids, as well as co-lipids were prepared and characterized. The liposomes were stable at 4°C and pH 7.4 for several months. Incubation with siRNA led to the full entrapment of nucleic acids as soon as the positive/negative charge ratio was superior to 2. The best liposomal formulation demonstrated a silencing efficiency up to 10% on HeLa cells, very similar to a commercial agent, with a lowest toxicity than the commercial agent. Using flow cytometry and microscopy assays, we demonstrated that drop of pH was required for the transfection efficiency, since bafilomycin blocked the transfection efficiency. Additional evidence was brought by the synthesis of a negative control lipid, which was unable to switch its conformation, and consequently exhibited no transfection ability. Mechanistic studies revealed that the uptake was mediated through endocytosis, by clathrin and caveolae pathways, as reported for previous lipid nanoparticle systems. This potent system was used for the treatment of hypercholesterolemia. The switchable lipids were able to knockdown PCSK9 expression on human hepatocytes (Huh-7). Its efficiency is currently evaluated on in vivo mice model of PCSK9 KO mice. In summary, we designed and optimized a new cationic pH-sensitive lipid for gene delivery. Its transfection efficiency is similar to the best available commercial agent, without the usually associated toxicity. The promising results lead to its use for the treatment of hypercholesterolemia on a mice model. Anticancer applications and pulmonary chronic disease are also currently investigated.

Keywords: liposomes, siRNA, pH-sensitive, molecular switch

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Authors: Shazia Anwer Bukhari, Madiha Javeed Ghani, Muhammad Ibrahim Rajoka

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Objective: Biochemical, environmental, physical and genetic factors have a strong effect on the development of coronary disease (CAD). Plasma homocysteine (Hcy) level and DNA damage play a pivotal role in its development and progression. The aim of this study was to investigate the predictive strength of an oxidative stress, clinical biomarkers and total antioxidant status (TAS) in CAD patients to find the correlation of homocysteine, TOS and oxidative DNA damage with other clinical parameters. Methods: Sixty confirmed patients with CAD and 60 healthy individuals as control were included in this study. Different clinical and laboratory parameters were studied in blood samples obtained from patients and control subjects using commercially available biochemical kits and statistical software Results: As compared to healthy individuals, CAD patients had significantly higher concentrations of indices of oxidative stress: homocysteine (P=0.0001), total oxidative stress (TOS) (P=0.0001), serum cholesterol (P=0.04), low density lipoprotein cholesterol (LDL) (P=0.01), high density lipoprotein-cholesterol (HDL) (P=0.0001), and malondialdehyde (MDA) (P=0.001) than those of healthy individuals. Plasma homocysteine level and oxidative DNA damage were positively correlated with cholesterol, triglycerides, systolic blood pressure, urea, total protein and albumin (P values= 0.05). Both Hcy and oxidative DNA damage were negatively correlated with TAS and proteins. Conclusion: Coronary artery disease patients had a significant increase in homocysteine level and DNA damage due to increased oxidative stress. In conclusion, our study shows a significantly increase in lipid peroxidation, TOS, homocysteine and DNA damage in the erythrocytes of patients with CAD. A significant decrease level of HDL-C and TAS was observed only in CAD patients. Therefore these biomarkers may be useful diagnosis of patients with CAD and play an important role in the pathogenesis of CAD.

Keywords: antioxidants, coronary artery disease, DNA damage, homocysteine, oxidative stress, malondialdehyde, 8-Hydroxy-2’deoxyguanosine

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Authors: Ataa A. Said, Elsayed A. Abuotabl, Gehan F. Abdel Raoof, Khaled Y. Mohamed

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The aim of this study is to evaluate antihyperglycaemic and antihyperlipidemic activities of Pleiogynium timorense (DC.) Leenh (Anacardiaceae) seeds as well as to isolate and identify the bioactive compounds. Antihyperglycaemic effect was evaluated by measuring the effect of two dose levels (150 and 300 mg/kg) of 70% methanol extract of Pleiogynium timorense seeds on blood glucose level when administered 45 minutes before glucose loading. In addition, the effect of the plant extract on the lipid profile was determined by measuring serum total lipids (TL), total cholesterol (TC), triglycerides (TG), high density lipoprotein cholesterol (HDL-C) and low density lipoprotein cholesterol (LDL-C). Furthermore, the bioactive compounds were isolated and identified by chromatographic and spectrometric methods.The results showed that the methanolic extract of the seeds significantly reduced the levels of blood glucose,(TL), (TC), (TG) and (LDL-C) but no significant effect on (HDL-C) comparing with control group. Furthermore, four phenolic compound were isolated which were identified as; catechin, gallic acid, para methoxy benzaldehyde and pyrogallol which were isolated for the first time from the plant. In addition sulphur -containing compound (sulpholane) was isolated for the first time from the plant and from the family. To our knowledge, this is the first study about antihyperglycaemicand antihyperlipidemic activities of the seeds of Pleiogyniumtimorense and its bioactive compounds. So, the methanolic extract of the seeds of Pleiogynium timorense could be a step towards the development of new antihyperglycaemic and antihyperlipidemic drugs.

Keywords: antihyperglycaemic, bioactive compounds, phenolic, Pleiogynium timorense, seeds

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Authors: Nabila N. El-Maraghy, Amany Essa, Yousra Abdel–Mottaleb, Nada Ismail

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The use of combination of chemotherapy and natural products to influence the cell death with low doses of chemotherapeutic agents and few side effects has recently emerged as a new method of cancer therapy. Aim: Evaluation the modulatory effect of Thymoquinone on HepG2 cells treated with Sorafenib. Methods: Hepatocellular Carcinoma HepG2 cell line was treated with Sorafenib and TQ individually and in combination. The effect of these treatments on cell viability (MTT assay), apoptosis (Expression of Caspase-3) and oxidative markers (GSH content and extent of lipid peroxidation) was determined. Results: When compared the effect of both agents alone and the combination of the IC50 of Sorafenib and the IC50 TQ, the combination resulted in reduction of cell inhibition and apoptosis and antagonize their actions on GSH content and extent of lipid peroxidation which are increased. This study showed potent anti-tumor activity of both TQ and Sorafenib separately on HepG2 but upon combination surprisingly they interacted and give antagonistic effect. Conclusion: Co-treatment resulted in antagonistic interaction between Sorafenib and Thymoquinone.

Keywords: antagonism, hepatocellular carcinoma, sorafenib, thymoquinone

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Authors: Patricia O. Carvalho, Marcia C. F. Messias, Salvador Sanchez Vinces, Caroline F. A. Gatinoni, Vitor P. Iordanu, Carlos A. R. Martinez

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Lipidomics methods are widely used in the identification and validation of disease-specific biomarkers and therapy response evaluation. The present study aimed to identify a panel of potential lipid biomarkers to evaluate response to neoadjuvant chemoradiotherapy in rectal adenocarcinoma (RAC). Liquid chromatography–mass spectrometry (LC-MS)-based untargeted lipidomic was used to profile human serum samples from patients with clinical stage T2 or T3 resectable RAC, after and before chemoradiotherapy treatment. A total of 28 blood plasma samples were collected from 14 patients with RAC who recruited at the São Francisco University Hospital (HUSF/USF). The study was approved by the ethics committee (CAAE 14958819.8.0000.5514). Univariate and multivariate statistical analyses were applied to explore dysregulated metabolic pathways using untargeted lipidic profiling and data mining approaches. A total of 36 statistically significant altered lipids were identified and the subsequent partial least-squares discriminant analysis model was both cross validated (R2, Q2) and permutated. Lisophosphatidyl-choline (LPC) plasmalogens containing palmitoleic and oleic acids, with high variable importance in projection score, showed a tendency to be lower after completion of chemoradiotherapy. Chemoradiotherapy seems to change plasmanyl-phospholipids levels, indicating that these lipids play an important role in the RAC pathogenesis.

Keywords: lipidomics, neoadjuvant chemoradiotherapy, plasmalogens, rectal adenocarcinoma

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Authors: Balunkeswar Nayak, Hanjun Cao, Xinruo Zhang

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Elderberry could protect human health from oxidative stress, and reduce aging and certain cardiovascular diseases due to the presence of bioactive phytochemicals with high antioxidant capacity. However, these bioactive phytochemicals, such as anthocyanins and other phenolic acids, are susceptible to degradation during processing of elderberries to juice, jam, and powder due to intensity and duration of thermal exposure. The effects of microwave heating rate during come-up-times, using a domestic 2450 MHz microwave, on the color, total anthocyanin content and total phenolics on elderberry juice was studied. With a variation of come-up-time from 30 sec to 15 min at different power levels (10–50 % of total wattage), the temperature of elderberry juice vary from 40.6 °C to 91.5 °C. However, the color parameters (L, A, and B), total anthocyanin content (using pH differential method) and total phenolics did not vary significantly when compared to the control samples.

Keywords: elderberry, microwave, color, thermal exposure

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Authors: Pedro Cabrales, Scott Caroen, Tony R. Reid, Bryan Oronsky

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Introduction and Purpose RRx-001 is an NLRP3 inhibitor and Nrf2 agonist in Phase 3 trials for the treatment of cancer. The purpose of this study was to examine whether treatment with RRx-001, given itsanti-inflammatory and antioxidant properties, improvedexercise and skeletal muscle oxidative capacity in mice on the generalpremiss that better health outcomes correlatewith more activity. Material and Methods Male and female adult mice (n=6 per group) were subjected to an endurance exercise capacity (EEC)test until exhaustion on a motorized treadmill after 3 once weekly doses of either RRx-001 5 mg/kg, RRx-001 2 mg/kg, or vehicle. The EEC protocol consisted of a treadmill velocity of 30meters per min at an uphill inclination (slope of 10%) until the mice reached fatigue, which was defined as the inability of the mice to maintain the appropriate pace despitecontinuous hand stimulation for 1 min. The concentration of malondialdehyde (MDA), an indicator of lipid peroxidation, and creatine kinase (CK), an indicator of muscle damage, in the blood samples collected immediately after the acute exercise was determined with a commercial ELISA assay kit. ResultsThe exhaustive exercise times of the RRx-001 groups were significantly longer than that of the vehicle group (p<0.05) by weeks 2 and 3. In addition, MDA levels in the gastrocnemius, soleus, and extensor digitorum longus muscles were significantly lower than those of the vehicle group were (p<0.05), as were the serum CK levels(p<0.05). ConclusionsIn conclusion, this study found that RRx-001 has anti-fatigue properties, as evidenced by an increase in exercise capacity with RRx-001 treatment, and protects against strenuous exercise-induced muscle damage and lipid peroxidation. This data potentially supports the use of RRx-001 in the clinic to improve exercise performance and reduce physical fatigue.

Keywords: RRx-001, anti-fatigue, muscle protection, increased exercise tolerance, lipid peroxidation

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Authors: Mae Genevieve G. Cheung, Michael G. Cuevas, Lovely Fe L. Cuison, Elijin P. Dai, Katrina Marie S. Duron, Azalea Damaris E. Encarnacion, May T. Magtoto, Gina C. Castro

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The present study aims to evaluate the effectiveness of aqueous and ethanolic leaf extracts of Blumea balsamifera in reducing obesity on diet-induced obese Sprague-Dawley rats. Aqueous and ethanolic leaf extracts were obtained by maceration and percolation, respectively, of air-dried, grinded leaves. The test animals were given a high fat diet (HFD) for 21 days, except for one negative control group fed with a standard diet (SD). The Blumea balsamifera extracts were given at doses of 300 mg/Kg and 600 mg/Kg for BBAE and BBEE groups, and the positive control group, Orlistat, was given at 21.6 mg/Kg dose. After 24 days of treatment, the statistical difference of parameters such as Lee’s index and lipid profile of each group before and after the treatment period were determined separately using Tukey’s test of two-way Analysis of Variance (ANOVA). The statistical results showed that the600mg/kg dose of BBAE and BBEE had greatly lowered the Lee’s index among the other doses while the 300 mg/Kg dose BBEE, 600 mg/Kg BBAE, and 300 mg/kg BBAE lowered the total cholesterol level, LDL level, and VLDL and total triglyceride level respectively. The extracts, however, lowered the HDL level which was also exhibited by the standard drug, Orlistat.

Keywords: adipocytes, adipogenesis, Blumea balsamifera, Lee’s index, obesity, Sambong

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Authors: Ibrahim I. Elshahawy

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The objective of the present study was to investigate the effect of green tea extract on serum oxidant and antioxidant profile, liver and kidney function. 40 Friesian calves are included in this study and allocated into two groups: Group I (n=20) clinically healthy calves showing no clinical abnormalities, not receiving any treatment and served as control; group II (n=20) received green tea extract (GTE) for 30 days. Non-significant changes in blood urea nitrogen (BUN) were detected between groups, on contrary, serum creatinine and activities of liver enzymes aspartate transaminase (AST) and alanine transaminase (ALT) were significantly different between two groups. There were significant increases in the mean values of serum antioxidative parameters (total antioxidant capacity, catalase, superoxide dismutase, reduced glutathione and glutathione peroxidase) in group II. Whereas, the activity of lipid peroxidase significantly decreased in GTE treated calves when compared to control.

Keywords: green tea extract, antioxidants, oxidants, calves

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Authors: Navid Kaboudi, Ali Shayanfar

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Introduction: Feeding infants with safe milk from the beginning of their life is an important issue. Drugs which are used by mothers can affect the composition of milk in a way that is not only unsuitable, but also toxic for infants. Consuming permeable drugs during that sensitive period by mother could lead to serious side effects to the infant. Due to the ethical restrictions of drug testing on humans, especially women, during their lactation period, computational approaches based on structural parameters could be useful. The aim of this study is to develop mechanistic models to predict the M/P ratio of drugs during breastfeeding period based on their structural descriptors. Methods: Two hundred and nine different chemicals with their M/P ratio were used in this study. All drugs were categorized into two groups based on their M/P value as Malone classification: 1: Drugs with M/P>1, which are considered as high risk 2: Drugs with M/P>1, which are considered as low risk Thirty eight chemical descriptors were calculated by ACD/labs 6.00 and Data warrior software in order to assess the penetration during breastfeeding period. Later on, four specific models based on the number of hydrogen bond acceptors, polar surface area, total surface area, and number of acidic oxygen were established for the prediction. The mentioned descriptors can predict the penetration with an acceptable accuracy. For the remaining compounds (N= 147, 158, 160, and 174 for models 1 to 4, respectively) of each model binary regression with SPSS 21 was done in order to give us a model to predict the penetration ratio of compounds. Only structural descriptors with p-value<0.1 remained in the final model. Results and discussion: Four different models based on the number of hydrogen bond acceptors, polar surface area, and total surface area were obtained in order to predict the penetration of drugs into human milk during breastfeeding period About 3-4% of milk consists of lipids, and the amount of lipid after parturition increases. Lipid soluble drugs diffuse alongside with fats from plasma to mammary glands. lipophilicity plays a vital role in predicting the penetration class of drugs during lactation period. It was shown in the logistic regression models that compounds with number of hydrogen bond acceptors, PSA and TSA above 5, 90 and 25 respectively, are less permeable to milk because they are less soluble in the amount of fats in milk. The pH of milk is acidic and due to that, basic compounds tend to be concentrated in milk than plasma while acidic compounds may consist lower concentrations in milk than plasma. Conclusion: In this study, we developed four regression-based models to predict the penetration class of drugs during the lactation period. The obtained models can lead to a higher speed in drug development process, saving energy, and costs. Milk/plasma ratio assessment of drugs requires multiple steps of animal testing, which has its own ethical issues. QSAR modeling could help scientist to reduce the amount of animal testing, and our models are also eligible to do that.

Keywords: logistic regression, breastfeeding, descriptors, penetration

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Authors: Essam Amerian

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The aim of this study was to investigate the effects of improving waist-to-hip ratio (WHR) and visceral fat components on the health of shift workers in an offshore petrochemical company. A total of 100 male shift workers participated in the study, with an average age of 40.5 years and an average BMI of 28.2 kg/m². The study employed a randomized controlled trial design, with participants assigned to either an intervention group or a control group. The intervention group received a 12-week program that included dietary counseling, physical activity recommendations, and stress management techniques. The control group received no intervention. The outcomes measured were changes in WHR, visceral fat components, blood pressure, and lipid profile. The results showed that the intervention group had a statistically significant improvement in WHR (p<0.001) and visceral fat components (p<0.001) compared to the control group. Furthermore, there were statistically significant improvements in systolic blood pressure (p=0.015) and total cholesterol (p=0.034) in the intervention group compared to the control group. These findings suggest that implementing a 12-week program that includes dietary counseling, physical activity recommendations, and stress management techniques can effectively improve WHR, visceral fat components, and cardiovascular health among shift workers in an offshore petrochemical company.

Keywords: body composition, waist-hip-ratio, visceral fat, shift worker, work efficiency

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Authors: Wongsakorn Charoenpanitseri

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The total chromatic number χ"(G) of a graph G is the minimum number of colors needed to color the elements (vertices and edges) of G such that no incident or adjacent pair of elements receive the same color Let G be a graph with maximum degree Δ(G). Considering a total coloring of G and focusing on a vertex with maximum degree. A vertex with maximum degree needs a color and all Δ(G) edges incident to this vertex need more Δ(G) + 1 distinct colors. To color all vertices and all edges of G, it requires at least Δ(G) + 1 colors. That is, χ"(G) is at least Δ(G) + 1. However, no one can find a graph G with the total chromatic number which is greater than Δ(G) + 2. The Total Coloring Conjecture states that for every graph G, χ"(G) is at most Δ(G) + 2. In this paper, we prove that the Total Coloring Conjectur for a Δ-claw-free 3-degenerated graph. That is, we prove that the total chromatic number of every Δ-claw-free 3-degenerated graph is at most Δ(G) + 2.

Keywords: total colorings, the total chromatic number, 3-degenerated, CLAW-FREE

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Authors: Abdel Rahman A. Alzandi

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The primary objectives of this investigation were to examine the interactive effects of three air quality treatments, ethylene-diurea (EDU) and two irrigation conditions on physiological characteristics of kidney beans (Phaseolus vulgaris L.) during its whole growth. These plants were grown in 12-open top chambers (OTC's). Ethylene-diurea (EDU) was used as a factor to evaluate O3 pollution impact on plant growth. The air quality treatments consisted of charcoal filtered (CF) air, nonfiltered (NF) air and ambient air (AA) were irrigated and non- irrigated. Leaf samples were collected from upper canopy positions six times (pre- EDU addition, week after four EDU's addition, at the time of harvesting). Maximal differences in leaf carbohydrate, N contents, pigments and total lipids were observed in response to moisture conditions in presence and absence of EDU applications. Significant reduction were noted for air quality treatments regarding carbohydrate and pigment fractions but not for all cases of leaf N and lipid contents under O3 effects only. Minimal differences were found for first EDU application while maximal ones were recorded at 200 mg l-1 of treatments. The EDU treatments stimulated carbohydrate and pigment contents at the upper canopy position with higher levels for both NF and AA compared to untreated conditions. The NF and AA treatments caused lower total carbohydrate and pigment contents in the canopy position before harvesting of EDU applications. The stimulation in leaf carbohydrates by the EDU treatment, compared to the non-treated EDU of AA and NF treatments, provides a rational explanation for the counteracting effects of EDU against moderate exposures to O3 regarding grain yields in C3 plants.

Keywords: leaf contents, moisture relations, EDU additions, global climate change, kidney bean

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Authors: Qi Liu, Jiqin Yao, Xiaohu Zhou, Bo Zheng

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The first artificial cell was produced by Thomas Chang in the 1950s when he was trying to make a mimic of red blood cells. Since then, many different types of artificial cells have been constructed from one of the two approaches: a so-called bottom-up approach, which aims to create a cell from scratch, and a top-down approach, in which genes are sequentially knocked out from organisms until only the minimal genome required for sustaining life remains. In this project, bottom-up approach was used to build a new cell-free expression system which mimics artificial cell that capable of protein expression and communicate with each other. The artificial cells constructed from the bottom-up approach are usually lipid vesicles, polymersomes, hydrogels or aqueous droplets containing the nucleic acids and transcription-translation machinery. However, lipid vesicles based artificial cells capable of communication present several issues in the cell communication research: (1) The lipid vesicles normally lose the important functions such as protein expression within a few hours. (2) The lipid membrane allows the permeation of only small molecules and limits the types of molecules that can be sensed and released to the surrounding environment for chemical communication; (3) The lipid vesicles are prone to rupture due to the imbalance of the osmotic pressure. To address these issues, the hydrogel-based artificial cells were constructed in this work. To construct the artificial cell, polyacrylamide hydrogel was functionalized with Acrylate PEG Succinimidyl Carboxymethyl Ester (ACLT-PEG2000-SCM) moiety on the polymer backbone. The proteinaceous factors can then be immobilized on the polymer backbone by the reaction between primary amines of proteins and N-hydroxysuccinimide esters (NHS esters) of ACLT-PEG2000-SCM, the plasmid template and ribosome were encapsulated inside the hydrogel particles. Because the artificial cell could continuously express protein with the supply of nutrients and energy, the artificial cell-artificial cell communication and artificial cell-natural cell communication could be achieved by combining the artificial cell vector with designed plasmids. The plasmids were designed referring to the quorum sensing (QS) system of bacteria, which largely relied on cognate acyl-homoserine lactone (AHL) / transcription pairs. In one communication pair, “sender” is the artificial cell or natural cell that can produce AHL signal molecule by synthesizing the corresponding signal synthase that catalyzed the conversion of S-adenosyl-L-methionine (SAM) into AHL, while the “receiver” is the artificial cell or natural cell that can sense the quorum sensing signaling molecule form “sender” and in turn express the gene of interest. In the experiment, GFP was first immobilized inside the hydrogel particle to prove that the functionalized hydrogel particles could be used for protein binding. After that, the successful communication between artificial cell-artificial cell and artificial cell-natural cell was demonstrated, the successful signal between artificial cell-artificial cell or artificial cell-natural cell could be observed by recording the fluorescence signal increase. The hydrogel-based artificial cell designed in this work can help to study the complex communication system in bacteria, it can also be further developed for therapeutic applications.

Keywords: artificial cell, cell-free system, gene circuit, synthetic biology

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Authors: Gian Carlo F. Maliwat, Stephanie F. Velasquez, Janice A. Ragaza

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A 50-day growth trial was conducted to evaluate the efficacy of Chlorella vulgaris (Beijerinck) as an ingredient in the diets of giant freshwater prawn Macrobrachium rosenbergii (De Man) postlarvae (PL30). Immune response (total haemocyte count and prophenoloxidase activity) was also assessed by subjecting postlarvae to a challenge test against Aeromonas hydrophila (Chester) for 14 days. Isonitrogenous and iso-lipidic test diets were prepared using a fishmeal-based-positive control diet (D0) and four basal diets with inclusion levels of 2% (D2), 4% (D4), 6% (D6) and 8% (D8) C. vulgaris. Postlarvae of M. rosenbergii were randomly stocked (mean initial body weight of 0.19 ± 0.02 g) in 30-L tanks in three replicates per dietary treatment for evaluation of growth performance. Another set of postlarvae (mean initial body weight of 1.25 ± 0.02 g) was randomly distributed in 95-L tanks in three replicates per dietary treatment for the assessment of immune response. Results showed that specific growth rate was significantly higher (P < 0.05) in postlarvae fed D4 and D6. Variations in values for carcass protein, lipid, moisture, and ash were also evident. Postlarvae fed diets with Chlorella showed increased prophenol oxidase activity and total haemocyte counts. Moreover, the survival rate after challenge with A. hydrophila was significantly increased (P < 0.05). Inclusion of C. vulgaris in diets enhanced immune response and resistance of M. rosenbergii postlarvae against A. hydrophila infection.

Keywords: Chlorella vulgaris, haemocyte count, Macrobrachium rosenbergii, prophenoloxidase activity

Procedia PDF Downloads 119

Authors: A. Mandić, M. Sakač, A. Mišan, B. Šojić, L. Petrović, I. Lončarević, B. Pajin, I. Sedej

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Aldehydes as secondary lipid oxidation products are highly specific to the oxidative degradation of particular polyunsaturated fatty acids present in foods. Gas chromatographic analysis of those volatile compounds has been widely used for monitoring of the deterioration of food products. Developed static headspace gas chromatography method using flame ionization detector (SHS GC FID) was applied to monitor the aldehydes present in processed foods such as bakery, meat and confectionary products. Five selected aldehydes were determined in samples without any sample preparation, except grinding for bakery and meat products. SHS–GC analysis allows the separation of propanal, pentanal, hexanal, heptanal and octanal, within 15min. Aldehydes were quantified in fresh and stored samples, and the obtained range of aldehydes in crackers was 1.62±0.05-9.95±0.05mg/kg, in sausages 6.62±0.46-39.16±0.39mg/kg; and in cocoa spread cream 0.48±0.01-1.13±0.02mg/kg. Referring to the obtained results, the following can be concluded, proposed method is suitable for different types of samples, content of aldehydes varies depending on the type of a sample, and differs in fresh and stored samples of the same type.

Keywords: lipid oxidation, aldehydes, crackers, sausage, cocoa cream spread

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Authors: Scott Nielsen, Luca Longanesi, Chris Chuck

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Palm oil is obtained from the flesh and kernel of the fruit of oil palms and is the most productive and inexpensive oil crop. The global demand for palm oil is approximately 75 million metric tonnes, a 29% increase in global production of palm oil since 2016. This expansion of oil palm cultivation has resulted in mass deforestation, vast biodiversity destruction and increasing net greenhouse gas emissions. One possible alternative is to produce a saturated oil, similar to palm, from microbes such as oleaginous yeast. The yeasts can be cultured on sugars derived from second-generation sources and do not compete with tropical forests for land. One highly promising oleaginous yeast for this application is Metschnikowia pulcherrima. However, recent techno-economic modeling has shown that cell lysis and standard lipid extraction are major contributors to the cost of the oil. Typical cell disruption techniques to extract either single cell oils or proteins have been based around bead-beating, homogenization and acid lysis. However, these can have a detrimental effect on lipid quality and are energy-intensive. In this study, a vortex separator, which produces high sheer with minimal energy input, was investigated as a potential low energy method of lysing cells. This was compared to four more traditional methods (thermal lysis, acid lysis, alkaline lysis, and osmotic lysis). For each method, the yeast loading was also examined at 1 g/L, 10 g/L and 100 g/L. The quality of the cell disruption was measured by optical cell density, cell counting and the particle size distribution profile comparison over a 2-hour period. This study demonstrates that the vortex separator is highly effective at lysing the cells and could potentially be used as a simple apparatus for lipid recovery in an oleaginous yeast process. The further development of this technology could potentially reduce the overall cost of microbial lipids in the future.

Keywords: palm oil substitute, metschnikowia pulcherrima, cell disruption, cell lysis

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Authors: Surendra Agrawal, Pravina Gurjar, Supriya Bhide, Ram Gaud

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Levamisole hydrochloride is a prominent anticancer drug in the treatment of colon cancer but resulted in toxic effects due poor bioavailability and poor cellular uptake by tumor cells. Levamisole is an unstable drug. Incorporation of this molecule in solid lipids may minimize their exposure to the aqueous environment and partly immobilize the drug molecules within the lipid matrix-both of which may protect the encapsulated drugs against degradation. The objectives of the study were to enhance bioavailability by sustaining drug release and to reduce the toxicities associated with the therapy. Solubility of the drug was determined in different lipids to select the components of Solid Lipid Nanoparticles (SLN). Pseudoternary phase diagrams were created using aqueous titration method. Formulations were subjected to particle size and stability evaluation to select the final test formulations which were characterized for average particle size, zeta potential, and in-vitro drug release and percentage transmittance to optimize the final formulation. SLN of Levamisole hydrochloride was prepared by Nanoprecipitation method. Glyceryl behenate (Compritol 888 ATO) was used as core comprising of Tween 80 as surfactant and Lecithin as co-surfactant in (1:1) ratio. Entrapment efficiency (EE) was found to be 45.89%. Particle size was found in the range of 100-600 nm. Zeta potential of the formulation was -17.0 mV revealing the stability of the product. In-vitro release study showed that 66 % drug released in 24 hours in pH 7.2 which represent that formulation can give controlled action at the intestinal environment. In pH 5.0 it showed 64% release indicating that it can even release drug in acidic environment of tumor cells. In conclusion, results revealed SLN to be a promising approach to sustain the drug release so as to increase bioavailability and cellular uptake of the drug with reduction in toxic effects as dose has been reduced with controlled delivery.

Keywords: SLN, nanoparticulate delivery of levamisole, pharmacy, pharmaceutical sciences

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