Search results for: lipid metabolism
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 1087

Search results for: lipid metabolism

247 Exploring the Design of Prospective Human Immunodeficiency Virus Type 1 Reverse Transcriptase Inhibitors through a Comprehensive Approach of Quantitative Structure Activity Relationship Study, Molecular Docking, and Molecular Dynamics Simulations

Authors: Mouna Baassi, Mohamed Moussaoui, Sanchaita Rajkhowa, Hatim Soufi, Said Belaaouad

Abstract:

The objective of this paper is to address the challenging task of targeting Human Immunodeficiency Virus type 1 Reverse Transcriptase (HIV-1 RT) in the treatment of AIDS. Reverse Transcriptase inhibitors (RTIs) have limitations due to the development of Reverse Transcriptase mutations that lead to treatment resistance. In this study, a combination of statistical analysis and bioinformatics tools was adopted to develop a mathematical model that relates the structure of compounds to their inhibitory activities against HIV-1 Reverse Transcriptase. Our approach was based on a series of compounds recognized for their HIV-1 RT enzymatic inhibitory activities. These compounds were designed via software, with their descriptors computed using multiple tools. The most statistically promising model was chosen, and its domain of application was ascertained. Furthermore, compounds exhibiting comparable biological activity to existing drugs were identified as potential inhibitors of HIV-1 RT. The compounds underwent evaluation based on their chemical absorption, distribution, metabolism, excretion, toxicity properties, and adherence to Lipinski's rule. Molecular docking techniques were employed to examine the interaction between the Reverse Transcriptase (Wild Type and Mutant Type) and the ligands, including a known drug available in the market. Molecular dynamics simulations were also conducted to assess the stability of the RT-ligand complexes. Our results reveal some of the new compounds as promising candidates for effectively inhibiting HIV-1 Reverse Transcriptase, matching the potency of the established drug. This necessitates further experimental validation. This study, beyond its immediate results, provides a methodological foundation for future endeavors aiming to discover and design new inhibitors targeting HIV-1 Reverse Transcriptase.

Keywords: QSAR, ADMET properties, molecular docking, molecular dynamics simulation, reverse transcriptase inhibitors, HIV type 1

Procedia PDF Downloads 92
246 Cost Effective and Efficient Feeding: A Way Forward for Sustainable and Profitable Aquaculture

Authors: Pawan Kumar Sharma, J. Stephan Sampath Kumar, S. Anand, Chandana B. L.

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Protein is the major component for the success in culture of shrimp and fishes. Apparently, excess dietary protein is undesirable, as it not only enhances the production cost but also leads to water quality deterioration. A field survey was conducted with aqua farmers of Kerala, India, a leading state in coastal aquaculture, to assess the role of protein component in feed that can be efficiently and effectively managed for sustainable aquaculture. The study showed an average feed amount of 13.55 ± 2.16 tonnes per hectare was being used by the farmers of Kerala. The average feed cost percentage of Rs. 57.76 ± 13.46 /kg was invested for an average protein level of 36.26 % ± 0.082 in the feed and Rs.78.95 ± 3.086 per kilogram of feed was being paid by the farmers. Study revealed that replacement of fish meal and fish oil within shrimp aquafeeds with alternative protein, and lipid sources can only be achieved if changes are made in the basic shrimp culturing practices, such as closed farming system through water recycling or zero-water exchange, and by maximizing in-situ, floc and natural food production within the culture system. The upshot of such production systems is that imports of high-quality feed ingredients and aqua feeds can eventually be eliminated, and the utilization of locally available feed ingredients from agricultural by-products can be greatly improved and maximized. The promotion of closed shrimp production systems would also greatly reduce water use and increase shrimp production per unit area but would necessitate the continuous provision of electricity for aeration during production. Alternative energy sources such as solar power might be used, and resource poor farming communities should also explore wind energy for use. The study concluded that farm made feed and closed farming systems are essential for the sustainability and profitability of the aquaculture industry.

Keywords: aqua feeds, floc, fish meal, protein, zero-water exchange

Procedia PDF Downloads 145
245 A Gold-Based Nanoformulation for Delivery of the CRISPR/Cas9 Ribonucleoprotein for Genome Editing

Authors: Soultana Konstantinidou, Tiziana Schmidt, Elena Landi, Alessandro De Carli, Giovanni Maltinti, Darius Witt, Alicja Dziadosz, Agnieszka Lindstaedt, Michele Lai, Mauro Pistello, Valentina Cappello, Luciana Dente, Chiara Gabellini, Piotr Barski, Vittoria Raffa

Abstract:

CRISPR/Cas9 technology has gained the interest of researchers in the field of biotechnology for genome editing. Since its discovery as a microbial adaptive immune defense, this system has been widely adopted and is acknowledged for having a variety of applications. However, critical barriers related to safety and delivery are persisting. Here, we propose a new concept of genome engineering, which is based on a nano-formulation of Cas9. The Cas9 enzyme was conjugated to a gold nanoparticle (AuNP-Cas9). The AuNP-Cas9 maintained its cleavage efficiency in vitro, to the same extent as the ribonucleoprotein, including non-conjugated Cas9 enzyme, and showed high gene editing efficiency in vivo in zebrafish embryos. Since CRISPR/Cas9 technology is extensively used in cancer research, melanoma was selected as a validation target. Cell studies were performed in A375 human melanoma cells. Particles per se had no impact on cell metabolism and proliferation. Intriguingly, the AuNP-Cas9 internalized spontaneously in cells and localized as a single particle in the cytoplasm and organelles. More importantly, the AuNP-Cas9 showed a high nuclear localization signal. The AuNP-Cas9, overcoming the delivery difficulties of Cas9, could be used in cellular biology and localization studies. Taking advantage of the plasmonic properties of gold nanoparticles, this technology could potentially be a bio-tool for combining gene editing and photothermal therapy in cancer cells. Further work will be focused on intracellular interactions of the nano-formulation and characterization of the optical properties.

Keywords: CRISPR/Cas9, gene editing, gold nanoparticles, nanotechnology

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244 Unveiling the Detailed Turn Off-On Mechanism of Carbon Dots to Different Sized MnO₂ Nanosensor for Selective Detection of Glutathione

Authors: Neeraj Neeraj, Soumen Basu, Banibrata Maity

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Glutathione (GSH) is one of the most important biomolecules having small molecular weight, which helps in various cellular functions like regulation of gene, xenobiotic metabolism, preservation of intracellular redox activities, signal transduction, etc. Therefore, the detection of GSH requires huge attention by using extremely selective and sensitive techniques. Herein, a rapid fluorometric nanosensor is designed by combining carbon dots (Cdots) and MnO₂ nanoparticles of different sizes for the detection of GSH. The bottom-up approach, i.e., microwave method, was used for the preparation of the water soluble and greatly fluorescent Cdots by using ascorbic acid as a precursor. MnO₂ nanospheres of different sizes (large, medium, and small) were prepared by varying the ratio of concentration of methionine and KMnO₄ at room temperature, which was confirmed by HRTEM analysis. The successive addition of MnO₂ nanospheres in Cdots results fluorescence quenching. From the fluorescence intensity data, Stern-Volmer quenching constant values (KS-V) were evaluated. From the fluorescence intensity and lifetime analysis, it was found that the degree of fluorescence quenching of Cdots followed the order: large > medium > small. Moreover, fluorescence recovery studies were also performed in the presence of GSH. Fluorescence restoration studies also show the order of turn on follows the same order, i.e., large > medium > small, which was also confirmed by quantum yield and lifetime studies. The limits of detection (LOD) of GSH in presence of Cdots@different sized MnO₂ nanospheres were also evaluated. It was observed thatLOD values were in μM region and lowest in case of large MnO₂ nanospheres. The separation distance (d) between Cdots and the surface of different MnO₂ nanospheres was determined. The d values increase with increase in the size of the MnO₂ nanospheres. In summary, the synthesized Cdots@MnO₂ nanocomposites acted as a rapid, simple, economical as well as environmental-friendly nanosensor for the detection of GSH.

Keywords: carbon dots, fluorescence, glutathione, MnO₂ nanospheres, turn off-on

Procedia PDF Downloads 152
243 Hyaluronic Acid - Alginate Hydrogel for the Transdifferentiation of Testis Cells into Erythrocyte and Hepatocyte-like Cells; A Practice Within an Effective Agent Choice

Authors: Leila Rashki Ghaleno, Mohamad Amin Hajari, Leila Montazeri, Abdolhossein Shahverdi, Mojtaba Rezazadeh Valojerdi

Abstract:

Background: Spermatogonia stem cells (SSCs) exhibit pluripotency, enabling them to undergo differentiation into many cell lineages, including neurons, glia, endothelial cells, and hepatocytes when cultured in vitro. Although the specific mechanisms are not yet fully understood, it has been observed that biopolymer agents, such as hyaluronic acid (HA) and alginate (Alg), have the potential to induce transdifferentiation of SSCs. The current work aimed to examine the process of in vitro spermatogenesis and the conversion of mouse testicular cells into hepatocytes and erythrocyte-like cells utilizing the HA-Alg hydrogel. Method: After being extracted from the testes of a 5-day postpartum mouse (5 DPP), the testicular cells were separated into two enzymatic stages and then put into a composite hydrogel containing 0.5% HA and 1% alginate. On days 14 and 28 of culture, the colonies' growth, the cells' viability, and their histology were assessed. Result: Despite observing significant cell proliferation on day 14 and the development of circular-shaped organoids on day 28, it was noted that the organoids generated in the HA-Alg medium tended to maintain their circular morphology on day 28. Notably, the testicular cells underwent transdifferentiation into cell types resembling erythrocytes and hepatocytes. The hepatocyte-like cells exhibited the presence of glycogen and lipid deposits, indicating their hepatocyte-like characteristics. Interestingly, immunostaining analysis revealed the secretion of albumin and the presence of VEGFR on day 14. However, on day 28, albumin expression was not detected, while the expression of Sox9 (a marker for hepatocytes), Vegf, CD34, and C-kit (markers for erythrocytes) showed increased levels in the gene expression evaluation. Conclusion: The present findings indicated that HA-Alg could be a potent and effective agent for the transdifferentiation of testis cells into erythrocyte and hepatocyte-like cells, as recent studies have confirmed the transformation of SSCs into hepatocyte cells during in vitro culture.

Keywords: 3D culture, mouse testicular cell, hyaluronic acid, liver organoids

Procedia PDF Downloads 71
242 Gene Expression Analysis for Corals / Zooxanthellae under High Seawater Temperature Stress

Authors: Haruka Ito, Toru Maruyama, Michihiro Ito, Chuya Shinzato, Hiroyuki Fujimura, Yoshikatsu Nakano, Shoichiro Suda, Sachiyo Aburatani, Haruko Takeyama

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Clarifying symbiotic relationships is one of the most important theme for understanding the marine eco-system. Coral reef has been regarded as an important environmental resource. Coral holobiont composed by coral, symbiotic microalgae zooxanthellae, and bacteria have complexed relationship. Zooxanthellae mainly supply organic matter to the host corals through their photosynthetic activity. The symbiotic relationship is indispensable for corals but may easily collapses due to the rise of seawater temperature. However, the molecular mechanism how seawater temperature influences their relationships still remain unclear. In this study, the transcriptomic analysis has applied to elucidate the coral-zooxanthellae relationships under high seawater temperature stress. To observe reactions of corals and zooxanthellae against the rise of seawater temperature, meta-gene expression in coral have been analyzed. The branches from six different colonies of a stony coral, Acropora tenuis, were sampled at nine times by 2016 at two locations, Ishikawabaru and South of Sesoko Island, Okinawa, Japan. The mRNAs extracted from the branches including zooxanthellae were sequenced by illumina HiSeq. Gene Set Enrichment Analysis (GSEA) based on hyper geometric distribution was performed. The seawater temperature at 2016 summer was unusually high, which was caused by El Niño event, and the number of zooxanthellae in coral was decreased in August. GSEA derived the several specific genes expressed in A. tenuis under heat stress conditions. The upregulated genes under heat stress highly related with infection immunity. The downregulated genes significantly contained cell cycle related genes. Thu, it is considered that heat stress cause disorder in cell metabolism of A. tenuis, resulting in serious influence to coral holobiont.

Keywords: coral, symbiosis, thermal stress response, transcriptome analysis

Procedia PDF Downloads 272
241 Effect of High Dose of Vitamin C in Reduction Serum Uric Acid: a Comparative Study between Hyperuricemic and Gouty Patients in Jeddah

Authors: Firas S. Azzeh

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Background: Vitamin C is a water soluble vitamin that is necessary for normal growth and development. Hyperuricemia is commonly detected in subjects with abnormal purine metabolism. Prolonged hyperuricemia is an important risk factor for damaged joint and often associated with gout. Objectives: To compare the effect of high dose of vitamin C supplements on uric acid treatment between hyperuricemic and gouty patients in Jeddah, Saudi Arabia, as well as finding out the effect of vitamin C on serum creatinine level and glomerular filtration rate (GFR). Subjects and Methods: This comparative study started on April 2013 and lasted tells March 2014. A convenience sample of 30 adults was recruited in this study from Doctor Abdulrahman Taha Bakhsh Hospital in Jeddah (Saudi Arabia). Eligible persons were assigned into two study groups; hyperuricemic (n=15) and gouty (n=15) groups. Subjects have been accepted for participating in the study after completing the consent form. Each participant consumed 500 mg/day vitamin C chew able tablets. All participants have been followed-up for 2 months. Twelve hours fasting blood samples have been collected 3 times from each participant during the study period; at the beginning before and retested after each month of the study period. Uric acid, serum creatinine and GFR were measured. Results: For gouty group, uric acid increased insignificantly after 2 months by about +0.3 mg/dl. On the other hand, hyperuricemic group showed decrease (P ≤ 0.05) in uric acid after 2 months of study period by about -0.78 mg/dl. Serum creatinine level insignificantly decreased for all participants during the study period, which leaded to insignificant increase in GFR for all participants. Conclusion: Supplementation with 500 mg/day vitamin C for 2 months significantly reduced serum uric acid for hyperuricemic patients and insignificantly increased serum uric acid for gouty patients. The ineffectiveness of vitamin C supplements on patients with established gout could be related to a number of potential reasons.

Keywords: vitamin c, Hyperuricemia, gout, creatinine, GFR

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240 Resveratrol-Phospholipid Complex for Sustained Delivery of Resveratrol via the Skin for the Treatment of Inflammatory Diseases

Authors: Malay K. Das, Bhupen Kalita

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The poor oral bioavailability of resveratrol (RSV) due to presystemic metabolism can be avoided via dermal route of administration. The hydrophilic-lipophilic nature of resveratrol-phospholipid complex (RSVPs) favors the delivery of resveratrol via the skin. The RSVPs embedded polymeric patch with moderate adhesiveness was developed for dermal application for sustained anti-inflammatory effect. The prepared patches were evaluated for various physicochemical properties, surface morphology by SEM, TEM, and compatibility of patch components by FT-IR and DSC studies. The dermal flux of the optimized patch formulation was found to be at 4.28 ± 0.48 mg/cm2/24 h. The analysis of skin extract after permeation study revealed the presence of resveratrol, which confirmed the localization of RSVPs in the skin. The stability of RSVPs in the polymeric patch and the physiologic environment was confirmed by FE-SEM studies on the patches after drug release and skin permeation studies. The RSVPs particles released from the polymer matrix maintaining the structural integrity and permeate the keratinized horney layer of skin. The optimized patch formulation showed sustained anti-inflammatory effect (84.10% inhibition of inflammation at 24 h) in carrageenan-induced rat paw edema model compared to marketed diclofenac sodium gel (39.58% inhibition of inflammation at 24 h). The CLSM study confirmed the localization of RSVPs for a longer period, thus enabling drug targeting to the dermis for sustained anti-inflammatory effect. Histological studies with phase contrast trinocular microscope suggested no alteration of skin integrity and no evidence of the presence of inflammatory cells after exposure to the permeants. The patch was found to be safe for skin application as evaluated by Draize method for skin irritation scoring in a rabbit model. These results suggest the therapeutic efficacy of the developed patch in both acute and chronic inflammatory diseases.

Keywords: resveratrol-phospholipid complex, skin delivery, sustained anti-inflammatory effect, inflammatory diseases, dermal patch

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239 Impact of Dietary Rumen Protected Choline on Transition Dairy Cows’ Productive Performance

Authors: Mohamed Ahmed Tony, Fayez Abaza

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The effects of a dietary supplement of rumen-protected choline on feed intake, milk yield, milk composition and some blood metabolites were evaluated in transition dairy cows. Forty multiparous cows were blocked into 20 pairs and then randomly allocated to either one of 2 treatments. The treatments were supplementation either with or without (control) rumen-protected choline. Treatments were applied from 2 weeks before and until 8 weeks after calving. Both groups received the same basal diet as total mixed ration. Additionally, 50 g of a rumen-protected choline supplement (25% rumen protected choline chloride) was added individually in the feed. Individual feed intake, milk yield, and body weight were recorded daily. Milk samples were analyzed weekly for fat, protein, and lactose content. Blood was sampled at week 2 before calving, d 1, d 4, d 7, d 10, week 2, week 3, and week 8 after calving. Glucose, triglycerids, nonesterified fatty acids, and β-hydroxybutyric acid in blood were analysed. The results revealed that choline supplementation increased DM intake from 16.5 to 18.0 kg/d and, hence, net energy intake from 99.2 to 120.5 MJ/d at the intercept of the lactation curve at 1 day in milk. Choline supplementation had no effect on milk yield, milk fat yield, or lactose yield. Milk protein yield was increased from 1.11 to 1.22 kg/d at the intercept of the lactation curve. Choline supplementation was associated with decreased milk fat concentration at the intercept of the lactation curve at 1 day in milking, but the effect of choline on milk fat concentration gradually decreased as lactation progressed. Choline supplementation decreased the concentration of blood triglycerids during the first 4 wk after parturition. Choline supplementation had no effect on energy-corrected milk yield, energy balance, body weight and body condition score. Results from this study suggest that fat metabolism in periparturient dairy cows is improved by choline supplementation during the transition period and this may potentially decrease the risk for metabolic disorders in the periparturient dairy cow.

Keywords: choline, dairy cattle, transition cow, triglycerids

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238 Comparison of Fatty Acids Composition of Three Commercial Fish Species Farmed in the Adriatic Sea

Authors: Jelka Pleadin, Greta Krešić, Tina Lešić, Ana Vulić, Renata Barić, Tanja Bogdanović, Dražen Oraić, Ana Legac, Snježana Zrnčić

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Fish has been acknowledged as an integral component of a well-balanced diet, providing a healthy source of energy, high-quality proteins, vitamins, essential minerals and, especially, n-3 long-chain polyunsaturated fatty acids (n-3 LC PUFA), mainly eicosapentaenoic acid (20:5 n-3 EPA), and docosahexaenoicacid, (22:6 n-3 DHA), whose pleiotropic effects in terms of health promotion and disease prevention have been increasingly recognised. In this study, the fatty acids composition of three commercially important farmed fish species: sea bream (Sparus aurata), sea bass (Dicentrarchus labrax) and dentex (Dentex dentex) was investigated. In total, 60 fish samples were retrieved during 2015 (n = 30) and 2016 (n = 30) from different locations in the Adriatic Sea. Methyl esters of fatty acids were analysed using gas chromatography (GC) with flame ionization detection (FID). The results show that the most represented fatty acid in all three analysed species is oleic acid (C18:1n-9, OA), followed by linoleic acid (C18:2n-6, LA) and palmitic acid (C16:0, PA). Dentex was shown to have two to four times higher eicosapentaenoic (EPA) and docosahexaenoic (DHA) acid content as compared to sea bream and sea bass. The recommended n-6/n-3 ratio was determined in all fish species but obtained results pointed to statistically significant differences (p < 0.05) in fatty acid composition among the analysed fish species and their potential as a dietary source of valuable fatty acids. Sea bass and sea bream had a significantly higher proportion of n-6 fatty acids, while dentex had a significantly higher proportion of n-3 (C18:4n-3, C20:4n-3, EPA, DHA) fatty acids. A higher hypocholesterolaemic and hypercholesterolaemic fatty acids (HH) ratio was determined for sea bass and sea bream, which comes as the consequence of a lower share of SFA determined in these two species in comparison to dentex. Since the analysed fish species vary in their fatty acids composition consumption of diverse fish species would be advisable. Based on the established lipid quality indicators, dentex, a fish species underutilised by the aquaculture, seems to be a highly recommendable and important source of fatty acids recommended to be included into the human diet.

Keywords: dentex, fatty acids, farmed fish, sea bass, sea bream

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237 Proteomic Analysis of the Inhibition of Prolyl Oligopeptidase Induced by Z-Pro-Prolinal in Filarial Parasites

Authors: Mohit Wadhawan, Sushma Rathaur

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Lymphatic filariasis, also called elephantiasis is a tropical disease afflicting over 120 million people in 81 countries worldwide. Existing anti filarial drugs are effective against the larval stages of filarial parasites which call for an urgent need of drugs which are macrofilaricidal. Identification of molecular targets crucial for survival of filarial parasites is a prerequisite for drug designing. Prolyl oligopeptidase (POP) is one such crucial enzyme involved in the maturation and degradation of neuropeptides and peptide hormones. We have identified this peptidase in the bovine filarial parasite, Setaria cervi. Effect of inhibition of POP on the proteome profile of filarial parasite has been discussed in this study. Filarial parasites were exposed to Z-pro-prolinal (ZPP), a specific POP inhibitor for 8 h and the motility and viability of the parasites was observed. It significantly reduced the motility and viability of the parasites. To study the proteome profile, the cytosolic, endoplasmic reticulum (ER) and mitochondrial extracts of the adult female parasites were subjected to 2-dimensional electrophoresis. As analyzed by the PD-Quest software, the ZPP caused the alteration in the different subcellular proteins, and the significantly altered proteins were identified using MALDI-MS/MS spectrometry. The major proteins identified were found to play important role in diverse biological functions like signaling, redox regulation, energy metabolism, stress response, and cytoskeleton formation. Moreover, we found upregulation in the calcium binding proteins such as calreticulin, calponin, and calpain-6 suggesting that POP inhibition regulates calcium release. This relates to earlier reports that POP plays non-catalytic role in inositol 1,4,5-trisphosphate (IP3) signaling inducing release of calcium from ER. Taken together, the data demonstrated that inhibition of prolyl oligopeptidase alter the overall proteome signifying its role in survival of the filarial parasites. Thus this study provides a basis for the use of POP as a chemotherapeutic target for the treatment of lymphatic filariasis.

Keywords: lymphatic filariasis, setaria cervi, prolyl oligopeptidase, proteomics

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236 Liposome Sterile Filtration Fouling: The Impact of Transmembrane Pressure on Performance

Authors: Hercules Argyropoulos, Thomas F. Johnson, Nigel B Jackson, Kalliopi Zourna, Daniel G. Bracewell

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Lipid encapsulation has become essential in drug delivery, notably for mRNA vaccines during the COVID-19 pandemic. However, their sterile filtration poses challenges due to the risk of deformation, filter fouling and product loss from adsorption onto the membrane. Choosing the right filtration membrane is crucial to maintain sterility and integrity while minimizing product loss. The objective of this study is to develop a rigorous analytical framework utilizing confocal microscopy and filtration blocking models to elucidate the fouling mechanisms of liposomes as a model system for this class of delivery vehicle during sterile filtration, particularly in response to variations in transmembrane pressure (TMP) during the filtration process. Experiments were conducted using fluorescent Lipoid S100 PC liposomes formulated by micro fluidization and characterized by Multi-Angle Dynamic Light Scattering. Dual-layer PES/PES and PES/PVDF membranes with 0.2 μm pores were used for filtration under constant pressure, cycling from 30 psi to 5 psi and back to 30 psi, with 5, 6, and 5-minute intervals. Cross-sectional membrane samples were prepared by microtome slicing and analyzed with confocal microscopy. Liposome characterization revealed a particle size range of 100-140 nm and an average concentration of 2.93x10¹¹ particles/mL. Goodness-of-fit analysis of flux decline data at varying TMPs identified the intermediate blocking model as most accurate at 30 psi and the cake filtration model at 5 psi. Membrane resistance analysis showed atypical behavior compared to therapeutic proteins, with resistance remaining below 1.38×10¹¹ m⁻¹ at 30 psi, increasing over fourfold at 5 psi, and then decreasing to 1-1.3-fold when pressure was returned to 30 psi. This suggests that increased flow/shear deforms liposomes enabling them to more effectively navigate membrane pores. Confocal microscopy indicated that liposome fouling mainly occurred in the upper parts of the dual-layer membrane.

Keywords: sterile filtration, membrane resistance, microfluidization, confocal microscopy, liposomes, filtration blocking models

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235 Preventive Effect of Stem Back Extracts of Coula edulis Baill. against High-Fat / High Sucrose Diet-Induced Insulin Resistance and Oxidative Stress in Rats

Authors: Eric Beyegue, Boris Azantza, Judith Laure Ngondi, Julius E. Oben

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Background: Insulin resistance (IR) and oxidative stress are associated with obesity, diabetes mellitus, and other cardio metabolic disorders. The aim of this study was to investigate the effect of Coula edulis extracts (CEE) on insulin resistance and oxidative stress markers in high-fat/high sucrose diet-induced insulin resistance in rats. Materials and Methods: Thirty male rats were divided into 6 groups of 5 rats each fed, received daily oral administration of CE extracts for 8 weeks as follows: Group 1 or negative control group, fed with standard diet (SD); Group 2 fed with high-fat/high sucrose diet (HFHS) only; Group3 fed with HFHS + CEAq 200; Group 4 fed with HFHS + CEAq 400; Group 5 fed with HFHS + CEEt 200; Group 6 fed with HFHS + CEEt 400. At the end of the experiment (8 weeks), animals were sacrificed plasma lipid profile, glucose, insulin, oxidative marker and digestive enzyme activities were measured. The homeostasis model assessment for insulin resistance (HOMA-IR) was determined. Results: Feedings with HFHS significantly (p < 0.01) induced plasma hyperglycaemia, hyperinsulinaemia, increased triglyceride, total cholesterol, and low-density lipoprotein levels, decreased high-density lipoprotein levels, alterations of α amylase, and glucose-6-phosphatase activities, and oxidative stress. Daily oral administration with CEE for eight weeks after insulin resistance induction had a hypolipidaemic action, antioxidative activities and modulated metabolic markers. Ethanolic extract at the higher dose had the best effect on body weight gain and insulin resistance, whereas aqueous extract showed the better activity on hyperlipidemia. Conclusion: These results suggest that CEAq and CEEt at 400mg/kg are promising complementary supplements that can be used to protect better from metabolic disorders associated with HFHS.

Keywords: Coula edulis Baill, high-fat / high sucrose diet, insulin resistance, oxidative stress

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234 An Alternative Semi-Defined Larval Diet for Rearing of Sand Fly Species Phlebotomus argentipes in Laboratory

Authors: Faizan Hassan, Seema Kumari, V. P. Singh, Pradeep Das, Diwakar Singh Dinesh

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Phlebotomus argentipes is an established vector for Visceral Leishmaniasis in Indian subcontinent. Laboratory colonization of Sand flies is imperative in research on vectors, which requires a proper diet for their larvae and adult growth that ultimately affects their survival and fecundity. In most of the laboratories, adult Sand flies are reared on rabbit blood feeding/artificial blood feeding and their larvae on fine grinded rabbit faeces as a sole source of food. Rabbit faeces are unhygienic, difficult to handle, high mites infestation as well as owing to bad odour which creates menacing to human users ranging from respiratory problems to eye infection and most importantly it does not full fill all the nutrients required for proper growth and development. It is generally observed that the adult emergence is very low in comparison to egg hatched, which may be due to insufficient food nutrients provided to growing larvae. To check the role of food nutrients on larvae survival and adult emergence, a high protein rich artificial diet for sand fly larvae were used in this study. The composition of artificial diet to be tested includes fine grinded (9 gm each) Rice, Pea nuts & Soyabean balls. These three food ingredients are rich source of all essential amino acids along with carbohydrate and minerals which is essential for proper metabolism and growth. In this study artificial food was found significantly more effective for larval development and adult emergence than rabbit faeces alone (P value >0.05). The weight of individual larvae was also found higher in test pots than the control. This study suggest that protein plays an important role in insect larvae development and adding carbohydrate will also enhances the fecundity of insects larvae.

Keywords: artificial food, nutrients, Phlebotomus argentipes, sand fly

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233 Beneficial Effects of Curcumin against Stress Oxidative and Mitochondrial Dysfunction Induced by Trinitrobenzene Sulphonic Acid in Colon

Authors: Souad Mouzaoui, Bahia Djerdjouri

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Oxidative stress is one of the main factors involved in the onset and chronicity of inflammatory bowel disease (IBD). In this study, we investigated the beneficial effects of a potent natural antioxidant, curcumin (Cur) on colitis and mitochondrial dysfunction in trinitrobenzene sulfonic acid (TNBS)-induced colitis in mice. Rectal instillation of the chemical irritant TNBS (30 mg kg-1) induced the disruption of distal colonic architecture and a massive inflammatory cells influx to the mucosa and submucosa layers. Under these conditions, daily administration of Cur (25 mg kg-1) efficiently decreased colitis scores in the inflamed distal colon by reducing leukocyte infiltrate as attested by reduced myeloperoxidase (MPO) activity. Moreover, the levels of nitrite, an end product of inducible NO synthase activity (iNOS) and malonyl dialdehyde (MDA), a marker of lipid peroxidation increased in a time depending manner in response to TNBS challenge. Conversely, the markers of the antioxidant pool, reduced glutathione (GSH) and catalase activity (CAT) were drastically reduced. Cur attenuated oxidative stress markers and partially restored CAT and GSH levels. Moreover, our results expanded the effect of Cur on TNBS-induced colonic mitochondrial dysfunction. In fact, TNBS induced mitochondrial swelling and lipids peroxidation. These events reflected in the opening of mitochondrial transition pore and could be an initial indication in the cascade process leading to cell death. TNBS inhibited also mitochondrial respiratory activity, caused overproduction of mitochondrial superoxide anion (O2-.) and reduced level of mitochondrial GSH. Nevertheless, Cur reduced the extent of mitochondrial oxidative stress induced by TNBS and restored colonic mitochondrial function. In conclusion, our results showed the critical role of oxidative stress in TNBS-induced colitis. They highlight the role of colonic mitochondrial dysfunction induced by TNBS, as a potential source of oxidative damages. Due to its potent antioxidant properties, Cur opens a promising therapeutic approach against oxidative inflammation in IBD.

Keywords: colitis, curcumin, mitochondria, oxidative stress, TNBS

Procedia PDF Downloads 253
232 Synthesis and Characterization of Polycaprolactone for the Delivery of Rifampicin

Authors: Evelyn Osehontue Uroro, Richard Bright, Jing Yang Quek, Krasimir Vasilev

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Bacterial infections have been a challenge both in the public and private sectors. The colonization of bacteria often occurs in medical devices such as catheters, heart valves, respirators, and orthopaedic implants. When biomedical devices are inserted into patients, the deposition of macromolecules such as fibrinogen and immunoglobin on their surfaces makes it easier for them to be prone to bacteria colonization leading to the formation of biofilms. The formation of biofilms on medical devices has led to a series of device-related infections which are usually difficult to eradicate and sometimes cause the death of patients. These infections require surgical replacements along with prolonged antibiotic therapy, which would incur additional health costs. It is, therefore, necessary to prevent device-related infections by inhibiting the formation of biofilms using intelligent technology. Antibiotic resistance of bacteria is also a major threat due to overuse. Different antimicrobial agents have been applied to microbial infections. They include conventional antibiotics like rifampicin. The use of conventional antibiotics like rifampicin has raised concerns as some have been found to have hepatic and nephrotoxic effects due to overuse. Hence, there is also a need for proper delivery of these antibiotics. Different techniques have been developed to encapsulate and slowly release antimicrobial agents, thus reducing host cytotoxicity. Examples of delivery systems are solid lipid nanoparticles, hydrogels, micelles, and polymeric nanoparticles. The different ways by which drugs are released from polymeric nanoparticles include diffusion-based release, elution-based release, and chemical/stimuli-responsive release. Polymeric nanoparticles have gained a lot of research interest as they are basically made from biodegradable polymers. An example of such a biodegradable polymer is polycaprolactone (PCL). PCL degrades slowly by hydrolysis but is often sensitive and responsive to stimuli like enzymes to release encapsulants for antimicrobial therapy. This study presents the synthesis of PCL nanoparticles loaded with rifampicin and the on-demand release of rifampicin for treating staphylococcus aureus infections.

Keywords: enzyme, Staphylococcus aureus, PCL, rifampicin

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231 Prevalence and Risk Factors Associated with Nutrition Related Non-Communicable Diseases in a Cohort of Males in the Central Province of Sri Lanka

Authors: N. W. I. A. Jayawardana, W. A. T. A. Jayalath, W. M. T. Madhujith, U. Ralapanawa, R. S. Jayasekera, S. A. S. B. Alagiyawanna, A. M. K. R. Bandara, N. S. Kalupahana

Abstract:

There is mounting evidence to the effect that dietary and lifestyle changes affect the incidence of non-communicable diseases (NCDs). This study was conducted to investigate the association of diet, physical activity, smoking, alcohol consumption and duration of sleep with overweight, obesity, hypertension and diabetes in a cohort of males from the Central Province of Sri Lanka. A total of 2694 individuals aged between 17 – 68 years (Mean = 31) were included in the study. Body Mass Index cutoff values for Asians were used to categorize the participants as normal, overweight and obese. The dietary data were collected using a food frequency questionnaire [FFQ] and data on the level of physical activity, smoking, alcohol consumption and sleeping hours were obtained using a self-administered validated questionnaire. Systolic and diastolic blood pressure, random blood glucose levels were measured to determine the incidence of hypertension and diabetes. Among the individuals, the prevalence of overweight and obesity were 34% and 16.4% respectively. Approximately 37% of the participants suffered from hypertension. Overweight and obesity were associated with older age men (P<0.0001), frequency of smoking (P=0.0434), alcohol consumption level (P=0.0287) and the quantity of lipid intake (P=0.0081). Consumption of fish (P=0.6983) and salty snacks (P=0.8327), sleeping hours (P=0.6847) and the level of physical activity were not significantly (P=0.3301) associated with the incidence of overweight and obesity. Based on the fitted model, only age was significantly associated with hypertension (P < 0.001). Further, age (P < 0.0001), sleeping hours (P=0.0953) and consumption of fatty foods (P=0.0930) were significantly associated with diabetes. Age was associated with higher odds of pre diabetes (OR:1.089;95% CI:1.053,1.127) and diabetes (OR:1.077;95% CI:1.055,1.1) whereas 7-8 hrs. of sleep per day was associated with lesser odds of diabetes (OR:0.403;95% CI:0.184,0.884). High prevalence of overweight, obesity and hypertension in working-age males is a threatening sign for this area. As this population ages in the future and urbanization continues, the prevalence of above risk factors will likely to escalate.

Keywords: age, males, non-communicable diseases, obesity

Procedia PDF Downloads 337
230 The Effect of Different Exercise Intensities on Plasma Endostatin in Healthy Volunteers

Authors: Inayat Shah, Muhammad Omar Malik, Ghareeb Alshuwaier, Ronald H. Baxendale

Abstract:

Background: The balance between angiogenesis and angiostasis is important in growth and developmental processes in the body. Angiogenic and angiostatic mediators control this balance. Endostatin is one of the prominent angiostatic mediators. The marked angiostatic effect of endostatin includes inhibiting endothelial cell migration, proliferation and apoptosis. Physical activity decreases the risk and development of many angiogenesis related health problems including atherosclerosis and numerous cancers. Physiological influences of different physical activities on plasma endostatin concentration are controversial and not completely clear. Moreover, correlation of physical characteristics and metabolic predictors during physical activity on circulating endostatin is indistinct and poorly speculated. The study aimed to determine the effects of mild, moderate and vigorous exercise on the concentration of endostatin in plasma. Methodology: 22 participants, 16 males (age = 30.6 ± 7.8 years) and 6 females (age = 26.5 ± 5 years) were recruited. Weekly session of different intensities exercise based on the predicted maximum heart of the participants [60%(low), 70% (moderate) and 80% (vigorous)] were carried out. The duration and work rate for each participant was determined through sub-maximal exercise. Standardization of the session was done on total energy expenditure of the participants per session. One pre exercise and two post exercise samples were taken at intervals of 10 and 60 minutes. Results: Pre-exercise mean endostatin was 101 ± 20 ng/dl. Low intensity exercise insignificantly decreased the endostatin concentration in plasma at 10 and 60 minutes 97 ± 20 ng/dl (p= 0.5), 98 ± 23 ng/dl (p= 0.8)). However, moderate (p= 0.022, 0.004) and vigorous intensities (p ≤ 0.001, 0.02) increased the endostatin concentrations significantly at both 10 and 60 minutes intervals respectively. The effects were not significantly influenced by gender, exercise mode (walking vs. running), components of exercise (HR, Speed, Gradients, distance, duration) or metabolism during exercise (VO₂ max, VCO₂, RER, energy expenditure, rate of carbohydrate or fats oxidation). Conclusion: Low intensity exercises did not influence endostatin concentration. However, moderate to high intensity exercises significantly increase endostatin concentration and may have potential benefits.

Keywords: angiogenesis, exercise, endostatin, physical activity

Procedia PDF Downloads 223
229 Exploring the Prebiotic Potential of Glucosamine

Authors: Shilpi Malik, Ramneek Kaur, Archita Gupta, Deepshikha Yadav, Ashwani Mathur, Manisha Singh

Abstract:

Glucosamine (GS) is the most abundant naturally occurring amino monosaccharide and is normally produced in human body via cellular glucose metabolism. It is regarded as the building block of cartilage matrix and is also an essential component of cartilage matrix repair mechanism. Besides that, it can also be explored for its prebiotic potential as many bacterial species are known to utilize the amino sugar by acquiring them to form peptidoglycans and lipopolysaccharides in the bacterial cell wall. Glucosamine can therefore be considered for its fermentation by bacterial species present in the gut. Current study is focused on exploring the potential of glucosamine as prebiotic. The studies were done to optimize considerable concentration of GS to reach GI tract and being fermented by the complex gut microbiota and food grade GS was added to various Simulated Fluids of Gastro-Intestinal Tract (GIT) such as Simulated Saliva, Gastric Fluid (Fast and Fed State), Colonic fluid, etc. to detect its degradation. Since it was showing increase in microbial growth (CFU) with time, GS was Further, encapsulated to increase its residential time in the gut, which exhibited improved resistance to the simulated Gut conditions. Moreover, prepared microspehres were optimized and characterized for their encapsulation efficiency and toxicity. To further substantiate the prebiotic activity of Glucosamine, studies were also performed to determine the effect of Glucosamine on the known probiotic bacterial species, i.e. Lactobacillus delbrueckii (MTCC 911) and Bifidobacteriumbifidum (MTCC 5398). Culture conditions for glucosamine will be added in MRS media in anaerobic tube at 0.20%, 0.40%, 0.60%, 0.80%, and 1.0%, respectively. MRS media without GS was included in this experiment as the control. All samples were autoclaved at 118° C for 15 min. Active culture was added at 5% (v/v) to each anaerobic tube after cooling to room temperature and incubated at 37° C then determined biomass and pH and viable count at incubation 18h. The experiment was completed in triplicate and the results were presented as Mean ± SE (Standard error).The experimental results are conclusive and suggest Glucosamine to hold prebiotic properties.

Keywords: gastro intestinal tract, microspheres, peptidoglycans, simulated fluid

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228 Modulation of Isoprenaline-Induced Myocardial Damage by Atorvastatin

Authors: Dalia Atallah, Lamiaa Ahmed, Hala Zaki, Mahmoud Khattab

Abstract:

Background: Isoprenaline (ISO) administration induces myocardial damage via oxidative stress and endothelial dysfunction. Atorvastatin (ATV) treatment improves both oxidative stress and endothelial dysfunction yet recent studies have reported a pro-oxidant effect upon ATV administration on both clinical and experimental studies. The present study was directed to investigate the effect of ATV pre-treatment and treatment on ISO-induced myocardial damage. Methods: Male rats were divided into five groups (n = 10). Rats were given ISO (5mg/kg/day, i.p.) for one week with or without ATV (10mg/kg/day, p.o.). ATV was given either as pre-treatment for one week before its co-administration with ISO for another week or as a treatment for two weeks at the end of the ISO administration. At the end of the experiment, the electrocardiographic examination was done and blood was isolated for the estimation of plasma creatine kinase MB (CK-MB) activity. Rats were then sacrificed and the whole ventricles were isolated for histological examination and the estimation of lipid peroxides as malondialdehyde (MDA) level, reduced glutathione (GSH) level, catalase activity, total nitrate-nitrite (NOx), as well as the estimation of both endothelial nitric oxide synthase (eNOS) and inducible nitric oxide synthase (iNOS) protein expression. Results: ISO-induced myocardial damage showed a significant elevation in ST segment, an increase in CK-MB activity, as well as increased oxidative stress biomarkers. Also, ISO-treated rats showed a significant decrease in myocardial NOx level and eNOS as well as degeneration in the myocardium. ATV pre-treatment didn’t show any protection to ISO-treated rats. On the other hand, ATV treatment showed a significant decrease in both the elevated ST wave and CK-MB activity. Moreover, ATV Treatment succeeded to improve oxidative stress biomarkers, tissue NOx, and eNOS protein expression, as well as amelioration of the histological alterations. Conclusion: Pre-treatment with ATV failed to protect against ISO-induced damage. This might suggest a synergistic pro-oxidant effect upon administration of the pro-oxidant ISO along with ATV as demonstrated by the increased oxidative stress and endothelial dysfunction. On the other side, ATV treatment succeeded to significantly improve oxidative stress biomarkers, endothelial dysfunction and myocardial degeneration.

Keywords: atorvastatin, endothelial dysfunction, isoprenaline, oxidative stress

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227 The Use of Ultrasound as a Safe and Cost-Efficient Technique to Assess Visceral Fat in Children with Obesity

Authors: Bassma A. Abdel Haleem, Ehab K. Emam, George E. Yacoub, Ashraf M. Salem

Abstract:

Background: Obesity is an increasingly common problem in childhood. Childhood obesity is considered the main risk factor for the development of metabolic syndrome (MetS) (diabetes type 2, dyslipidemia, and hypertension). Recent studies estimated that among children with obesity 30-60% will develop MetS. Visceral fat thickness is a valuable predictor of the development of MetS. Computed tomography and dual-energy X-ray absorptiometry are the main techniques to assess visceral fat. However, they carry the risk of radiation exposure and are expensive procedures. Consequently, they are seldom used in the assessment of visceral fat in children. Some studies explored the potential of ultrasound as a substitute to assess visceral fat in the elderly and found promising results. Given the vulnerability of children to radiation exposure, we sought to evaluate ultrasound as a safer and more cost-efficient alternative for measuring visceral fat in obese children. Additionally, we assessed the correlation between visceral fat and obesity indicators such as insulin resistance. Methods: A cross-sectional study was conducted on 46 children with obesity (aged 6–16 years). Their visceral fat was evaluated by ultrasound. Subcutaneous fat thickness (SFT), i.e., the measurement from the skin-fat interface to the linea alba, and visceral fat thickness (VFT), i.e., the thickness from the linea alba to the aorta, were measured and correlated with anthropometric measures, fasting lipid profile, homeostatic model assessment for insulin resistance (HOMA-IR) and liver enzymes (ALT). Results: VFT assessed via ultrasound was found to strongly correlate with the BMI, HOMA-IR with AUC for VFT as a predictor of insulin resistance of 0.858 and cut off point of >2.98. VFT also correlates positively with serum triglycerides and serum ALT. VFT correlates negatively with HDL. Conclusions: Ultrasound, a safe and cost-efficient technique, could be a useful tool for measuring the abdominal fat thickness in children with obesity. Ultrasound-measured VFT could be an appropriate prognostic factor for insulin resistance, hypertriglyceridemia, and elevated liver enzymes in obese children.

Keywords: metabolic syndrome, pediatric obesity, sonography, visceral fat

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226 Identification of Some Factors Influencing Serum Uric Acid Concentration in Individuals With Metabolic Syndrome

Authors: Munkhtuul G., Bolortsetseg Z., Lutzul M., Sugar N., Nyamdorj D., Nomundari B., Zesemdorj O., Erdenebayar N., Lkhagvasuren T. S., Munkhbayarlakh S., Bayasgalan T. Uurtuya S. H.

Abstract:

Background: Elevated serum uric acid (SUA) levels are observed in metabolic and cardiovascular conditions as an early predictor of metabolic syndrome (MS). Hyperuricemia, characterised by high uric acid levels in serum, increases the risk of developing MS by 1.6 times. Being overweight and obese significantly contributes to developing MS and cardiovascular disorders. In Mongolia, the prevalence of overweight and obesity is reaching 48.8% among individuals aged 15 to 49 years, indicating a potential surge in the incidence of MS, cardiovascular disorders, diabetes mellitus, and gout.Objective: This study aimed to determine the SUA levels in men diagnosed with MS and investigate the factors influencing these levels.Methods: A total of 119 men aged 30-60, who underwent preventive examinations and resided in Ulaanbaatar city, were included in the study. The criteria established by the International Diabetes Federation (IDF), American Heart Association (AHA), and the National Heart, Lung, and Blood Institute (NHLBI) were employed to define metabolic syndrome. Hyperuricemia was defined as SUA levels ≥7 mg/dL. Dietary intake was evaluated through the 24-hour recall method.Results: The study revealed that the prevalence of MS among the participants was 42.9% (n=51), with hyperuricemia observed in 16.8% (n=20) of the individuals. Among men diagnosed with MS, 21.3% (n=10) exhibited hyperuricemia. The mean SUA levels were as follows: 4.7±0.8 mg/dL in the healthy group, 5.9±1.1 mg/dL in men without MS but presenting central obesity, and 6.2±1.3 mg/dL in men with MS. After adjusting for age and body mass index (BMI), a positive correlation was observed between SUA levels and triglycerides (β=0.93) as well as lipid accumulation product (LAP) (β=0.92) in men with MS. In the central obesity group, SUA levels exhibited a positive correlation with triglycerides (β=0.91), visceral adiposity index (VAI) (β=0.73), LAP (β=0.92), and cardiometabolic index (CMI) (β=0.69). The risk of hyperuricemia increased by 3.29 times with elevated triglycerides and 3.53 times with an increased LAP.Conclusion: The findings indicate that abdominal fat accumulation, as indicated by elevated triglyceride levels and LAP, is associated with increased SUA levels in men with MS. However, no significant relationship was observed between SUA levels and dietary intake.

Keywords: central obesity, obesity, triglycerides, hyperuricemia

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225 Ability of Bentonite-lactobacillus Rhamnosus GAF06 Mixture to Mitigate Aflatoxin M1 Damages in Balb/C Mice

Authors: Amina Aloui, Jalila Ben Salah-Abbès, Abdellah Zinedine, Amar Riba, Noel Durand, Catherine Brabet, Didier Montet, Samir Abbès

Abstract:

Mycotoxin contamination of food and feed-isa globaconcern, both economically and for public health. Aflatoxin M1 (AFM1) is the principal hydroxylated metabolite of aflatoxin B1. It is frequently found in milk and other dairy products. It is responsible for the development of hepatocellular carcinoma and immunotoxic in humans and animals. The reduction of its bioavailabilitybecomesa great demand in order to protect human and animal health. The use of probiotic bacteria and clay are demonstrated to be able to bind AFM1 in vitro. This study aimed to investigate, in vivo, the activity of two-component mixture: L. rhamnosusGAF06 (LR) and bentonite for reducing the oxidative stress and the histological alterationsinduced by AFM1 in the liver andkidneys. For the experiment, male mice were divided into 7 groups (6 mice/group) and treated, orally, by AFM1, alone or in combination with LR and/or bentonite, for 10 days as follows: group 1 control, group 2 treated with LR alone (2.108 CFU/ml), group 3 treated with bentonite alone (1g/kg), group 4 treated with AFM1 alone (100μg/kg), group 5 co-treated with LR+AFM1, group 6 co-treated with bentonite+AFM1, group 7 co-treated with bentonite+LR+AFM1. At the end of the treatment, the mice were sacrificed, and the livers and kidneys were collected for histological assays. Intracellular antioxidant activities and lipid peroxidation were also studied. The results showed that AFM1causeddamage in liver and kidney tissues, being evidence of hepatotoxicity and nephrotoxicity marked by necrotic cells. It increased the MDA level and decreased the antioxidant enzyme activities (SOD) in both organs. In contrast, the co-treatment with AFM1 plus LR and/or bentonitesignificantly improved the hepatic and renal tissues, regulated kidney, and liver antioxidant enzyme activities. This improvement was more remarkable with the administration of LR-bentonite mixture with AFM1.LR and bentonite alone showed to be safe during the treatment. This mixture can be a promising candidate for future applications in biotechnological processes that aimed to detoxify AFM1in food and feed.

Keywords: aflatoxin M1, bentonite, L. rhamnosus GAF06, oxidative stress, prevention

Procedia PDF Downloads 194
224 Identification of Candidate Congenital Heart Defects Biomarkers by Applying a Random Forest Approach on DNA Methylation Data

Authors: Kan Yu, Khui Hung Lee, Eben Afrifa-Yamoah, Jing Guo, Katrina Harrison, Jack Goldblatt, Nicholas Pachter, Jitian Xiao, Guicheng Brad Zhang

Abstract:

Background and Significance of the Study: Congenital Heart Defects (CHDs) are the most common malformation at birth and one of the leading causes of infant death. Although the exact etiology remains a significant challenge, epigenetic modifications, such as DNA methylation, are thought to contribute to the pathogenesis of congenital heart defects. At present, no existing DNA methylation biomarkers are used for early detection of CHDs. The existing CHD diagnostic techniques are time-consuming and costly and can only be used to diagnose CHDs after an infant was born. The present study employed a machine learning technique to analyse genome-wide methylation data in children with and without CHDs with the aim to find methylation biomarkers for CHDs. Methods: The Illumina Human Methylation EPIC BeadChip was used to screen the genome‐wide DNA methylation profiles of 24 infants diagnosed with congenital heart defects and 24 healthy infants without congenital heart defects. Primary pre-processing was conducted by using RnBeads and limma packages. The methylation levels of top 600 genes with the lowest p-value were selected and further investigated by using a random forest approach. ROC curves were used to analyse the sensitivity and specificity of each biomarker in both training and test sample sets. The functionalities of selected genes with high sensitivity and specificity were then assessed in molecular processes. Major Findings of the Study: Three genes (MIR663, FGF3, and FAM64A) were identified from both training and validating data by random forests with an average sensitivity and specificity of 85% and 95%. GO analyses for the top 600 genes showed that these putative differentially methylated genes were primarily associated with regulation of lipid metabolic process, protein-containing complex localization, and Notch signalling pathway. The present findings highlight that aberrant DNA methylation may play a significant role in the pathogenesis of congenital heart defects.

Keywords: biomarker, congenital heart defects, DNA methylation, random forest

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223 Comparative Functional Analysis of Two Major Sterol-Biosynthesis Regulating Transcription Factors, Hob1 and Sre1, in Pathogenic Cryptococcus Species Complex

Authors: Dong-Gi Lee, Suyeon Cha, Yong-Sun Bahn

Abstract:

Sterol lipid is essential for cell membrane structure in eukaryotic cells. In mammalian cells, sterol regulatory element binding proteins (SREBPs) act as principal regulators of cellular cholesterol which is essential for proper cell membrane fluidity and structure. SREBP and sterol regulation are related to levels of cellular oxygen because it is a major substrate for sterol synthesis. Upon cellular sterol and oxygen levels are depleted, SREBP is translocated to the Golgi where it undergoes proteolytic cleavage of N terminus, then it travels to the nucleus to play a role as transcription factor. In yeast cells, synthesis of ergosterol is also highly oxygen consumptive, and Sre1 is a transcription factor known to play a central role in adaptation to growth under low oxygen condition and sterol homeostasis in Cryptococcus neoformans. In this study, we observed phenotypes in other strains of Cryptococcus species by constructing hob1Δ and sre1Δ mutants to confirm whether the functions of both genes are conserved in most serotypes. As a result, hob1Δ showed no noticeable phenotype under treatment of antifungal drugs and most environmental stresses in R265 (C. gattii) and XL280 (C. neoformans), suggesting that Hob1 is related to sterol regulation only in H99 (serotype A). On the other hand, the function of Sre1 was found to be conserved in most serotypes. Furthermore, mating experiment of hob1Δ or sre1Δ showed dramatic defects in serotype A (H99) and D (XL280). It revealed that Hob1 and Sre1 related to mating ability in Cryptococcus species, especially cell fusion efficiency. In conclusion, HOB1 and SRE1 play crucial role in regulating sterol-homeostasis and differentiation in C. neoformans, moreover, Hob1 is specific gene in Cryptococcus neoformans. It suggests that Hob1 is considered as potent factor-targeted new safety antifungal drug.

Keywords: cryptococcus neoformans, Hob1, Sre1, sterol regulatory element binding proteins

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222 Comparison of 18F-FDG and 11C-Methionine PET-CT for Assessment of Response to Neoadjuvant Chemotherapy in Locally Advanced Breast Carcinoma

Authors: Sonia Mahajan Dinesh, Anant Dinesh, Madhavi Tripathi, Vinod Kumar Ramteke, Rajnish Sharma, Anupam Mondal

Abstract:

Background: Neo-adjuvant chemotherapy plays an important role in treatment of breast cancer by decreasing the tumour load and it offers an opportunity to evaluate response of primary tumour to chemotherapy. Standard anatomical imaging modalities are unable to accurately reflect the response to chemotherapy until several cycles of drug treatment have been completed. Metabolic imaging using tracers like 18F-fluorodeoxyglucose (FDG) as a marker of glucose metabolism or amino acid tracers like L-methyl-11C methionine (MET) have potential role for the measurement of treatment response. In this study, our objective was to compare these two PET tracers for assessment of response to neoadjuvant chemotherapy, in locally advanced breast carcinoma. Methods: In our prospective study, 20 female patients with histology proven locally advanced breast carcinoma underwent PET-CT imaging using FDG and MET before and after three cycles of neoadjuvant chemotherapy (CAF regimen). Thereafter, all patients were taken for MRM and the resected specimen was sent for histo-pathological analysis. Tumour response to the neoadjuvant chemotherapy was evaluated by PET-CT imaging using PERCIST criteria and correlated with histological results. Responses calculated were compared for statistical significance using paired t- test. Results: Mean SUVmax for primary lesion in FDG PET and MET PET was 15.88±11.12 and 5.01±2.14 respectively (p<0.001) and for axillary lymph nodes was 7.61±7.31 and 2.75±2.27 respectively (p=0.001). Statistically significant response in primary tumour and axilla was noted on both FDG and MET PET after three cycles of NAC. Complete response in primary tumour was seen in only 1 patient in FDG and 7 patients in MET PET (p=0.001) whereas there was no histological complete resolution of tumor in any patient. Response to therapy in axillary nodes noted on both PET scans were similar (p=0.45) and correlated well with histological findings. Conclusions: For the primary breast tumour, FDG PET has a higher sensitivity and accuracy than MET PET and for axilla both have comparable sensitivity and specificity. FDG PET shows higher target to background ratios so response is better predicted for primary breast tumour and axilla. Also, FDG-PET is widely available and has the advantage of a whole body evaluation in one study.

Keywords: 11C-methionine, 18F-FDG, breast carcinoma, neoadjuvant chemotherapy

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221 The Role of Nutrition and Food Engineering in Promoting Sustainable Food Systems

Authors: Sara Khan Mohammadi

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The world is facing a major challenge of feeding a growing population while ensuring the sustainability of food systems. The United Nations estimates that the global population will reach 9.7 billion by 2050, which means that food production needs to increase by 70% to meet the demand. However, this increase in food production should not come at the cost of environmental degradation, loss of biodiversity, and climate change. Therefore, there is a need for sustainable food systems that can provide healthy and nutritious food while minimizing their impact on the environment. Nutrition and Food Engineering: Nutrition and food engineering play a crucial role in promoting sustainable food system. Nutrition is concerned with the study of nutrients in foods, their absorption, metabolism, and their effects on health. Food engineering involves the application of engineering principles to design, develop, and optimize food processing operations. Together, nutrition and food engineering can help to create sustainable food systems by: 1. Developing Nutritious Foods: Nutritionists and food engineers can work together to develop foods that are rich in nutrients such as vitamins, minerals, fiber, and protein. These foods can be designed to meet the nutritional needs of different populations while minimizing waste. 2. Reducing Food Waste: Food waste is a major problem globally as it contributes to greenhouse gas emissions and wastes resources such as water and land. Nutritionists and food engineers can work together to develop technologies that reduce waste during processing, storage, transportation, and consumption. 3. Improving Food Safety: Unsafe foods can cause illnesses such as diarrhea, cholera, typhoid fever among others which are major public health concerns globally. Nutritionists and food engineers can work together to develop technologies that improve the safety of foods from farm to fork. 4. Enhancing Sustainability: Sustainable agriculture practices such as conservation agriculture can help reduce soil erosion while improving soil fertility. Nutritionists and food engineers can work together to develop technologies that promote sustainable agriculture practices.

Keywords: sustainable food, developing food, reducing food waste, food safety

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220 System Dietadhoc® - A Fusion of Human-Centred Design and Agile Development for the Explainability of AI Techniques Based on Nutritional and Clinical Data

Authors: Michelangelo Sofo, Giuseppe Labianca

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In recent years, the scientific community's interest in the exploratory analysis of biomedical data has increased exponentially. Considering the field of research of nutritional biologists, the curative process, based on the analysis of clinical data, is a very delicate operation due to the fact that there are multiple solutions for the management of pathologies in the food sector (for example can recall intolerances and allergies, management of cholesterol metabolism, diabetic pathologies, arterial hypertension, up to obesity and breathing and sleep problems). In this regard, in this research work a system was created capable of evaluating various dietary regimes for specific patient pathologies. The system is founded on a mathematical-numerical model and has been created tailored for the real working needs of an expert in human nutrition using the human-centered design (ISO 9241-210), therefore it is in step with continuous scientific progress in the field and evolves through the experience of managed clinical cases (machine learning process). DietAdhoc® is a decision support system nutrition specialists for patients of both sexes (from 18 years of age) developed with an agile methodology. Its task consists in drawing up the biomedical and clinical profile of the specific patient by applying two algorithmic optimization approaches on nutritional data and a symbolic solution, obtained by transforming the relational database underlying the system into a deductive database. For all three solution approaches, particular emphasis has been given to the explainability of the suggested clinical decisions through flexible and customizable user interfaces. Furthermore, the system has multiple software modules based on time series and visual analytics techniques that allow to evaluate the complete picture of the situation and the evolution of the diet assigned for specific pathologies.

Keywords: medical decision support, physiological data extraction, data driven diagnosis, human centered AI, symbiotic AI paradigm

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219 Effect of a Muscarinic Antagonist Drug on Extracellular Lipase Activityof Pseudomonas aeruginosa

Authors: Zohreh Bayat, Dariush Minai-Tehrani

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Pseudomonas aeruginosa is a Gram-negative, rode shape and aerobic bacterium that has shown to be resistance to many antibiotics. This resistance makes the bacterium very harmful in some diseases. It can also generate diseases in any part of the gastrointestinal tract from oropharynx to rectum. P. aeruginosa has become an important cause of infection, especially in patients with compromised host defense mechanisms. One of the most important reasons that make P. aeruginosa an emerging opportunistic pathogen in patients is its ability to use various compounds as carbon sources. Lipase is an enzyme that catalyzes the hydrolysis of lipids. Most lipases act at a specific position on the glycerol backbone of lipid substrate. Some lipases are expressed and secreted by pathogenic organisms during the infection. Muscarinic antagonist used as an antispasmodic and in urinary incontinence. The drug has little effect on glandular secretion or the cardiovascular system. It does have some local anesthetic properties and is used in gastrointestinal, biliary, and urinary tract spasms. Aim: In this study the inhibitory effect of a muscarinic antagonist on lipase of P. aeruginosa was investigated. Methods: P. aeruginosa was cultured in minimal salt medium with 1% olive oil as carbon source. The cells were harvested and the supernatant, which contained lipase, was used for enzyme assay. Results: Our results showed that the drug can inhibit P. aeruginosa lipase by competitive manner. In the presence of different concentrations of the drug, the Vmax (2 mmol/min/mg protein) of enzyme did not change, while the Km raised by increasing the drug concentration. The Ki (inhibition constant) and IC50 (the half maximal inhibitory concentration) value of drug was estimated to be about 30 uM and 60 uM which determined that the drug binds to enzyme with high affinity. Maximum activity of the enzyme was observed at pH 8 in the absence and presence of muscarinic antagonist, respectively. The maximum activity of lipase was observed at 600C and the enzyme became inactive at 900C. Conclusion: The muscarinic antagonist drug could inhibit lipase of P. aeruginosa and changed the kinetic parameters of the enzyme. The drug binded to enzyme with high affinity and did not chang the optimum pH of the enzyme. Temperature did not affect the binding of drug to musmuscarinic antagonist.

Keywords: Pseudomonas aeruginosa, drug, enzyme, inhibition

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218 Candida antartica Lipase Assisted Enrichment of n-3 PUFA in Indian Sardine Oil

Authors: Prasanna Belur, P. R. Ashwini, Sampath Charanyaa, I. Regupathi

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Indian oil sardine (Sardinella longiceps) are one of the richest and cheapest sources of n-3 polyunsaturated fatty acids (n-3 PUFA) such as Eicosapentaenoic acid (EPA) and Docosahexaenoic acid (DHA). The health benefits conferred by n-3 PUFA upon consumption, in the prevention and treatment of coronary, neuromuscular, immunological disorders and allergic conditions are well documented. Natural refined Indian Sardine oil generally contain about 25% (w/w) n-3 PUFA along with various unsaturated and saturated fatty acids in the form of mono, di, and triglycerides. Having high concentration of n-3 PUFA content in the glyceride form is most desirable for human consumption to avail maximum health benefits. Thus, enhancing the n-3 PUFA content while retaining it in the glyceride form with green technology is the need of the hour. In this study, refined Indian Sardine oil was subjected to selective hydrolysis by Candida antartica lipase to enhance n-3 PUFA content. The degree of hydrolysis and enhancement of n-3 PUFA content was estimated by determining acid value, Iodine value, EPA and DHA content (by Gas Chromatographic methods after derivitization) before and after hydrolysis. Various reaction parameters such as pH, temperature, enzyme load, lipid to aqueous phase volume ratio and incubation time were optimized by conducting trials with one parameter at a time approach. Incubating enzyme solution with refined sardine oil with a volume ratio of 1:1, at pH 7.0, for 60 minutes at 50 °C, with an enzyme load of 60 mg/ml was found to be optimum. After enzymatic treatment, the oil was subjected to refining to remove free fatty acids and moisture content using previously optimized refining technology. Enzymatic treatment at the optimal conditions resulted in 12.11 % enhancement in Degree of hydrolysis. Iodine number had increased by 9.7 % and n-3 PUFA content was enhanced by 112 % (w/w). Selective enhancement of n-3 PUFA glycerides, eliminating saturated and unsaturated fatty acids from the oil using enzyme is an interesting preposition as this technique is environment-friendly, cost effective and provide natural source of n-3 PUFA rich oil.

Keywords: Candida antartica, lipase, n-3 polyunsaturated fatty acids, sardine oil

Procedia PDF Downloads 233