Search results for: lipoate protein ligase B (LipB)
Commenced in January 2007
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Edition: International
Paper Count: 2296

Search results for: lipoate protein ligase B (LipB)

1756 Possible Mechanism of DM2 Development in OSA Patients Mediated via Rev-Erb-Alpha and NPAS2 Proteins

Authors: Filip Franciszek Karuga, Szymon Turkiewicz, Marta Ditmer, Marcin Sochal, Piotr Białasiewicz, Agata Gabryelska

Abstract:

Circadian rhythm, an internal coordinator of physiological processes is composed of a set of semi-autonomous clocks. Clocks are regulated through the expression of circadian clock genes which form feedback loops, creating an oscillator. The primary loop consists of activators: CLOCK, BMAL1 and repressors: CRY, PER. CLOCK can be substituted by the Neuronal PAS Domain Protein 2 (NPAS2). Orphan nuclear receptor (REV-ERB-α) is a component of the secondary major loop, modulating the expression of BMAL1. Circadian clocks might be disrupted by the obstructive sleep apnea (OSA), which has also been associated with type II diabetes mellitus (DM2). Interestingly, studies suggest that dysregulation of NPAS2 and REV-ERB-α might contribute to the pathophysiology of DM2 as well. The goal of our study was to examine the role of NPAS2 and REV-ERB-α in DM2 in OSA patients. After examination of the clinical data, all participants underwent polysomnography (PSG) to assess their apnea-hypopnea index (AHI). Based on the acquired data participants were assigned to one of 3 groups: OSA (AHI>30, no DM2; n=17 for NPAS2 and 34 for REV-ERB-α), DM2 (AHI>30 + DM2; n=7 for NPAS2 and 15 for REV-ERB-α) and control group (AHI<5, no DM2; n=16 for NPAS2 and 31 for REV-ERB-α). ELISA immunoassay was performed to assess the serum protein level of REV-ERB-α and NPAS2. The only statistically significant difference between groups was observed in NPAS2 protein level (p=0.037). Post-hoc analysis showed significant differences between the OSA and the control group (p=0.017). AHI and NPAS2 level was significantly correlated (r=-0.478, p=0.002) in all groups. A significant correlation was observed between the REV-ERB-α level and sleep efficiency (r=0.617, p=0.005) as well as sleep maintenance efficiency (r=0.645, p=0.003) in the OSA group. We conclude, that NPAS2 is associated with OSA severity and might contribute to metabolic sequelae of this disease. REV-ERB-α on the other hand can influence sleep continuity and efficiency.

Keywords: OSA, diabetes mellitus, endocrinology, chronobiology

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1755 An Effective Modification to Multiscale Elastic Network Model and Its Evaluation Based on Analyses of Protein Dynamics

Authors: Weikang Gong, Chunhua Li

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Dynamics plays an essential role in function exertion of proteins. Elastic network model (ENM), a harmonic potential-based and cost-effective computational method, is a valuable and efficient tool for characterizing the intrinsic dynamical properties encoded in biomacromolecule structures and has been widely used to detect the large-amplitude collective motions of proteins. Gaussian network model (GNM) and anisotropic network model (ANM) are the two often-used ENM models. In recent years, many ENM variants have been proposed. Here, we propose a small but effective modification (denoted as modified mENM) to the multiscale ENM (mENM) where fitting weights of Kirchhoff/Hessian matrixes with the least square method (LSM) is modified since it neglects the details of pairwise interactions. Then we perform its comparisons with the original mENM, traditional ENM, and parameter-free ENM (pfENM) on reproducing dynamical properties for the six representative proteins whose molecular dynamics (MD) trajectories are available in http://mmb.pcb.ub.es/MoDEL/. In the results, for B-factor prediction, mENM achieves the best performance among the four ENM models. Additionally, it is noted that with the weights of the multiscale Kirchhoff/Hessian matrixes modified, interestingly, the modified mGNM/mANM still has a much better performance than the corresponding traditional ENM and pfENM models. As to dynamical cross-correlation map (DCCM) calculation, taking the data obtained from MD trajectories as the standard, mENM performs the worst while the results produced by the modified mENM and pfENM models are close to those from MD trajectories with the latter a little better than the former. Generally, ANMs perform better than the corresponding GNMs except for the mENM. Thus, pfANM and the modified mANM, especially the former, have an excellent performance in dynamical cross-correlation calculation. Compared with GNMs (except for mGNM), the corresponding ANMs can capture quite a number of positive correlations for the residue pairs nearly largest distances apart, which is maybe due to the anisotropy consideration in ANMs. Furtherly, encouragingly the modified mANM displays the best performance in capturing the functional motional modes, followed by pfANM and traditional ANM models, while mANM fails in all the cases. This suggests that the consideration of long-range interactions is critical for ANM models to produce protein functional motions. Based on the analyses, the modified mENM is a promising method in capturing multiple dynamical characteristics encoded in protein structures. This work is helpful for strengthening the understanding of the elastic network model and provides a valuable guide for researchers to utilize the model to explore protein dynamics.

Keywords: elastic network model, ENM, multiscale ENM, molecular dynamics, parameter-free ENM, protein structure

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1754 Increasing Photosynthetic H2 Production by in vivo Expression of Re-Engineered Ferredoxin-Hydrogenase Fusion Protein in the Green Alga Chlamydomonas reinhardtii

Authors: Dake Xiong, Ben Hankamer, Ian Ross

Abstract:

The most urgent challenge of our time is to replace the depleting resources of fossil fuels by sustainable environmentally friendly alternatives. Hydrogen is a promising CO2-neutral fuel for a more sustainable future especially when produced photo-biologically. Hydrogen can be photosynthetically produced in unicellular green alga like Chlamydomonas reinhardtii, catalysed by the inducible highly active and bidirectional [FeFe]-hydrogenase enzymes (HydA). However, evolutionary and physiological constraints severely restrict the hydrogen yield of algae for industrial scale-up, mainly due to its competition among other metabolic pathways on photosynthetic electrons. Among them, a major challenge to be resolved is the inferior competitiveness of hydrogen production (catalysed by HydA) with NADPH production (catalysed by ferredoxin-NADP+-reductase (FNR)), which is essential for cell growth and takes up ~95% of photosynthetic electrons. In this work, the in vivo hydrogen production efficiency of mutants with ferredoxin-hydrogenase (Fd*-HydA1*) fusion protein construct, where the electron donor ferredoxin (Fd*) is fused to HydA1* and expressed in the model organism C. reinhardtii was investigated. Once Fd*-HydA1* fusion gene is expressed in algal cells, the fusion enzyme is able to draw the redistributed photosynthetic electrons and use them for efficient hydrogen production. From preliminary data, mutants with Fd*-HydA1* transgene showed a ~2-fold increase in the photosynthetic hydrogen production rate compared with its parental strain, which only possesses the native HydA in vivo. Therefore, a solid method of having more efficient hydrogen production in microalgae can be achieved through the expression of the synthetic enzymes.

Keywords: Chlamydomonas reinhardtii, ferredoxin, fusion protein, hydrogen production, hydrogenase

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1753 Utilization of Silk Waste as Fishmeal Replacement: Growth Performance of Cyprinus carpio Juveniles Fed with Bombyx mori Pupae

Authors: Goksen Capar, Levent Dogankaya

Abstract:

According to the circular economy model, resource productivity should be maximized and wastes should be reduced. Since earth’s natural resources are continuously depleted, resource recovery has gained great interest in recent years. As part of our research study on the recovery and reuse of silk wastes, this paper focuses on the utilization of silkworm pupae as fishmeal replacement, which would replace the original fishmeal raw material, namely the fish itself. This, in turn, would contribute to sustainable management of wild fish resources. Silk fibre is secreted by the silkworm Bombyx mori in order to construct a 'room' for itself during its transformation process from pupae to an adult moth. When the cocoons are boiled in hot water, silk fibre becomes loose and the silk yarn is produced by combining thin silk fibres. The remaining wastes are 1) sericin protein, which is dissolved in water, 2) remaining part of cocoon, including the dead body of B. mori pupae. In this study, an eight weeks trial was carried out to determine the growth performance of common carp juveniles fed with waste silkworm pupae meal (SWPM) as a replacement for fishmeal (FM). Four isonitrogenous diets (40% CP) were prepared replacing 0%, 33%, 50%, and 100% of the dietary FM with non-defatted silkworm pupae meal as a dietary protein source for experiments in C. carpio. Triplicate groups comprising of 20 fish (0.92±0.29 g) were fed twice/day with one of the four diets. Over a period of 8 weeks, results showed that the diet containing 50% of its protein from SWPM had significantly higher (p ≤ 0.05) growth rates in all groups. The increasing levels of SWPM were resulted in a decrease in growth performance and significantly lower growth (p ≤ 0.05) was observed with diets having 100% SWPM. The study demonstrates that it is practical to replace 50% of the FM protein with SWPM with a significantly better utilization of the diet but higher SWPM levels are not recommended for juvenile carp. Further experiments are under study to have more detailed results on the possible effects of this alternative diet on the growth performance of juvenile carp.

Keywords: Bombyx mori, Cyprinus carpio, fish meal, silk, waste pupae

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1752 Induction of G1 Arrest and Apoptosis in Human Cancer Cells by Panaxydol

Authors: Dong-Gyu Leem, Ji-Sun Shin, Sang Yoon Choi, Kyung-Tae Lee

Abstract:

In this study, we focused on the anti-proliferative effects of panaxydol, a C17 polyacetylenic compound derived from Panax ginseng roots, against various human cancer cells. We treated with panaxydol to various cancer cells and panaxydol treatment was found to significantly inhibit the proliferation of human lung cancer cells (A549) and human pancreatic cancer cells (AsPC-1 and MIA PaCa-2), of which AsPC-1 cells were most sensitive to its treatment. DNA flow cytometric analysis indicated that panaxydol blocked cell cycle progression at the G1 phase in A549 cells, which accompanied by a parallel reduction of protein expression of cyclin-dependent kinase (CDK) 2, CDK4, CDK6, cyclin D1 and cyclin E. CDK inhibitors (CDKIs), such as p21CIP1/WAF1 and p27KIP1, were gradually upregulated after panaxydol treatment at the protein levels. Furthermore, panaxydol induced the activation of p53 in A549 cells. In addition, panaxydol also induced apoptosis of AsPC-1 and MIA PaCa-2 cells, as shown by accumulation of subG1 and apoptotic cell populations. Panaxydol triggered the activation of caspase-3, -8, -9 and the cleavage of poly (ADP-ribose) polymerase (PARP). Reduction of mitochondrial transmembrane potential by panaxydol was determined by staining with dihexyloxacarbocyanine iodide. Furthermore, panaxydol suppressed the levels of anti-apoptotic proteins, XIAP and Bcl-2, and increased the levels of proapoptotic proteins, Bax and Bad. In addition, panaxydol inhibited the activation of Akt and extracellular signal-regulated kinase (ERK) and activated the p38 mitogen-activated protein kinase kinase (MAPK). Our results suggest that panaxydol is an anti-tumor compound that causes p53-mediated cell cycle arrest and apoptosis via mitochondrial apoptotic pathway in various cancer cells.

Keywords: apoptosis, cancer, G1 arrest, panaxydol

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1751 Clay Hydrogel Nanocomposite for Controlled Small Molecule Release

Authors: Xiaolin Li, Terence Turney, John Forsythe, Bryce Feltis, Paul Wright, Vinh Truong, Will Gates

Abstract:

Clay-hydrogel nanocomposites have attracted great attention recently, mainly because of their enhanced mechanical properties and ease of fabrication. Moreover, the unique platelet structure of clay nanoparticles enables the incorporation of bioactive molecules, such as proteins or drugs, through ion exchange, adsorption or intercalation. This study seeks to improve the mechanical and rheological properties of a novel hydrogel system, copolymerized from a tetrapodal polyethylene glycol (PEG) thiol and a linear, triblock PEG-PPG-PEG (PPG: polypropylene glycol) α,ω-bispropynoate polymer, with the simultaneous incorporation of various amounts of Na-saturated, montmorillonite clay (MMT) platelets (av. lateral dimension = 200 nm), to form a bioactive three-dimensional network. Although the parent hydrogel has controlled swelling ability and its PEG groups have good affinity for the clay platelets, it suffers from poor mechanical stability and is currently unsuitable for potential applications. Nanocomposite hydrogels containing 4wt% MMT showed a twelve-fold enhancement in compressive strength, reaching 0.75MPa, and also a three-fold acceleration in gelation time, when compared with the parent hydrogel. Interestingly, clay nanoplatelet incorporation into the hydrogel slowed down the rate of its dehydration in air. Preliminary results showed that protein binding by the MMT varied with the nature of the protein, as horseradish peroxidase (HRP) was more strongly bound than bovine serum albumin. The HRP was no longer active when bound, presumably as a result of extensive structural refolding. Further work is being undertaken to assess protein binding behaviour within the nanocomposite hydrogel for potential diabetic wound healing applications.

Keywords: hydrogel, nanocomposite, small molecule, wound healing

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1750 In Silico Study of Antiviral Drugs Against Three Important Proteins of Sars-Cov-2 Using Molecular Docking Method

Authors: Alireza Jalalvand, Maryam Saleh, Somayeh Behjat Khatouni, Zahra Bahri Najafi, Foroozan Fatahinia, Narges Ismailzadeh, Behrokh Farahmand

Abstract:

Object: In the last two decades, the recent outbreak of Coronavirus (SARS-CoV-2) imposed a global pandemic in the world. Despite the increasing prevalence of the disease, there are no effective drugs to treat it. A suitable and rapid way to afford an effective drug and treat the global pandemic is a computational drug study. This study used molecular docking methods to examine the potential inhibition of over 50 antiviral drugs against three fundamental proteins of SARS-CoV-2. METHODS: Through a literature review, three important proteins (a key protease, RNA-dependent RNA polymerase (RdRp), and spike) were selected as drug targets. Three-dimensional (3D) structures of protease, spike, and RdRP proteins were obtained from the Protein Data Bank. Protein had minimal energy. Over 50 antiviral drugs were considered candidates for protein inhibition and their 3D structures were obtained from drug banks. The Autodock 4.2 software was used to define the molecular docking settings and run the algorithm. RESULTS: Five drugs, including indinavir, lopinavir, saquinavir, nelfinavir, and remdesivir, exhibited the highest inhibitory potency against all three proteins based on the binding energies and drug binding positions deduced from docking and hydrogen-bonding analysis. Conclusions: According to the results, among the drugs mentioned, saquinavir and lopinavir showed the highest inhibitory potency against all three proteins compared to other drugs. It may enter laboratory phase studies as a dual-drug treatment to inhibit SARS-CoV-2.

Keywords: covid-19, drug repositioning, molecular docking, lopinavir, saquinavir

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1749 C-eXpress: A Web-Based Analysis Platform for Comparative Functional Genomics and Proteomics in Human Cancer Cell Line, NCI-60 as an Example

Authors: Chi-Ching Lee, Po-Jung Huang, Kuo-Yang Huang, Petrus Tang

Abstract:

Background: Recent advances in high-throughput research technologies such as new-generation sequencing and multi-dimensional liquid chromatography makes it possible to dissect the complete transcriptome and proteome in a single run for the first time. However, it is almost impossible for many laboratories to handle and analysis these “BIG” data without the support from a bioinformatics team. We aimed to provide a web-based analysis platform for users with only limited knowledge on bio-computing to study the functional genomics and proteomics. Method: We use NCI-60 as an example dataset to demonstrate the power of the web-based analysis platform and data delivering system: C-eXpress takes a simple text file that contain the standard NCBI gene or protein ID and expression levels (rpkm or fold) as input file to generate a distribution map of gene/protein expression levels in a heatmap diagram organized by color gradients. The diagram is hyper-linked to a dynamic html table that allows the users to filter the datasets based on various gene features. A dynamic summary chart is generated automatically after each filtering process. Results: We implemented an integrated database that contain pre-defined annotations such as gene/protein properties (ID, name, length, MW, pI); pathways based on KEGG and GO biological process; subcellular localization based on GO cellular component; functional classification based on GO molecular function, kinase, peptidase and transporter. Multiple ways of sorting of column and rows is also provided for comparative analysis and visualization of multiple samples.

Keywords: cancer, visualization, database, functional annotation

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1748 Investigation of Self-Assembling of Maghemite Nanoparticles into Chain–Like Structures Using Birefringence Measurements

Authors: C. R. Stein; K. Skeff Neto, K. L. C. Miranda, P. P. C. Sartoratto, M. E. Xavier, Z. G. M. Lacava, S. M. De Freita, P. C. Morais

Abstract:

In this study, static magnetic birefringence (SMB) and transmission electron microscopy (TEM) were used to investigate the self-assembling of maghemite nanoparticles suspended as biocompatible magnetic fluid (BMF) while incubated or not with the Black Eyed–Pea Trypsin Chymotripsin Inhibitor–BTCI protein. The stock samples herein studied are dextran coated maghemite nanoparticles (average core diameter of 7.1 nm, diameter dispersion of 0.26, and containing 4.6×1016 particle/mL) and the dextran coated maghemite nanoparticles associated with the BTCI protein. Several samples were prepared by diluting the stock samples with deionized water while following their colloidal stability. The diluted samples were investigated using SMB measurements to assess the average sizes of the self-assembled and suspended mesoscopic structures whereas the TEM micrographs provide the morphology of the as-suspended units. The SMB data were analyzed using a model that includes the particle-particle interaction within the mean field model picture.

Keywords: biocompatible magnetic fluid, maghemite nanoparticles, self-assembling

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1747 Contribution of NLRP3 Inflammasome to the Protective Effect of 5,14-HEDGE, A 20-HETE Mimetic, against LPS-Induced Septic Shock in Rats

Authors: Bahar Tunctan, Sefika Pinar Kucukkavruk, Meryem Temiz-Resitoglu, Demet Sinem Guden, Ayse Nihal Sari, Seyhan Sahan-Firat, Mahesh P. Paudyal, John R. Falck, Kafait U. Malik

Abstract:

We hypothesized that 20-hydroxyeicosatetraenoic acid (20-HETE) mimetics such as N-(20-hydroxyeicosa-5[Z],14[Z]-dienoyl)glycine (5,14-HEDGE) may be beneficial for preventing mortality due to inflammation induced by lipopolysaccharide (LPS). This study aims to assess the effect of 5,14-HEDGE on the LPS-induced changes in nucleotide binding domain and leucine-rich repeat protein 3 (NLRP3)/apoptosis-associated speck-like protein containing a caspase activation and recruitment domain (ASC)/pro-caspase-1 inflammasome. Rats were injected with saline (4 ml/kg) or LPS (10 mg/kg) at time 0. Blood pressure and heart rate were measured using a tail-cuff device. 5,14-HEDGE (30 mg/kg) was administered to rats 1 h after injection of saline or LPS. The rats were sacrificed 4 h after saline or LPS injection and kidney, heart, thoracic aorta, and superior mesenteric artery were isolated for measurement of caspase-1/11 p20, NLRP3, ASC, and β-actin proteins as well as interleukin-1β (IL-1β) levels. Blood pressure decreased by 33 mmHg and heart rate increased by 63 bpm in the LPS-treated rats. In the LPS-treated rats, tissue protein expression of caspase-1/11 p20, NLRP3, and ASC in addition to IL-1β levels were increased. 5,14-HEDGE prevented the LPS-induced changes. Our findings suggest that inhibition of renal, cardiac, and vascular formation/activity of NLRP3/ASC/pro-caspase-1 inflammasome involved in the protective effect of 5,14-HEDGE on LPS-induced septic shock in rats. This work was financially supported by the Mersin University (2015-AP3-1343) and USPHS NIH (PO1 HL034300).

Keywords: 5, 14-HEDGE, lipopolysaccharide, NLRP3, inflammasome, septic shock

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1746 A Deletion in Duchenne Muscular Dystrophy Gene Found Through Whole Exome Sequencing in Iran

Authors: Negin Parsamanesh, Saman Ameri-Mahabadi, Ali Nikfar, Mojdeh Mansouri, Hossein Chiti, Gita Fatemi Abhari

Abstract:

Duchenne muscular dystrophy (DMD) is a severe progressive X-linked neuromuscular illness that affects movement through mutations in dystrophin gene. The mutation leads to insufficient, lack of or dysfunction of dystrophin. The cause of DMD was determined in an Iranian family. Exome sequencing was carried out along with a complete physical examination of the family. In silico methods were applied to find the alteration in the protein structure. The homozygous variant in DMD gene (NM-004006.2) was defined as c.2732-2733delTT (p.Phe911CysfsX8) in exon 21. In addition, phylogenetic conservation study of the human dystrophin protein sequence revealed that phenylalanine 911 is one of the evolutionarily conserved amino acids. In conclusion, our study indicated a new deletion in the DMD gene in the affected family. This deletion with an X-linked inheritance pattern is new in Iran. These findings could facilitate genetic counseling for this family and other patients in the future.

Keywords: duchenne muscular dystrophy, whole exome sequencing, iran, metabolic syndrome

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1745 Lucilia Sericata Netrin-A: Secreted by Salivary Gland Larvae as a Potential to Neuroregeneration

Authors: Hamzeh Alipour, Masoumeh Bagheri, Tahereh Karamzadeh, Abbasali Raz, Kourosh Azizi

Abstract:

Netrin-A, a protein identified for conducting commissural axons, has a similar role in angiogenesis. In addition, studies have shown that one of the netrin-A receptors is expressed in the growing cells of small capillaries. It will be interesting to study this new group of molecules because their role in wound healing will become clearer in the future due to angiogenesis. The greenbottle blowfly Luciliasericata (L. sericata) larvae are increasingly used in maggot therapy of chronic wounds. This aim of this was the identification of moleculareatures of Netrin-A in L. sericata larvae. Larvae were reared under standard maggotarium conditions. The nucleic acid sequence of L. sericataNetrin-A (LSN-A) was then identified using Rapid Amplification of cDNA Ends (RACE) and Rapid Amplification of Genomic Ends (RAGE). Parts of the Netrin-A gene, including the middle, 3′-, and 5′-ends were identified, TA cloned in pTG19 plasmid, and transferred into DH5ɑ Escherichia coli. Each part was sequenced and assembled using SeqMan software. This gene structure was further subjected to in silico analysis. The DNA of LSN-A was identified to be 2407 bp, while its mRNA sequence was recognized as 2115 bp by Oligo0.7 software. It translated the Netrin-A protein with 704 amino acid residues. Its molecular weight is estimated to be 78.6 kDa. The 3-D structure ofNetrin-A drawn by SWISS-MODEL revealed its similarity to the Netrin-1 of humans with 66.8% identity. The LSN-A protein conduces to repair the myelin membrane in neuronal cells. Ultimately, it can be an effective candidate in neural regeneration and wound healing. Furthermore, our next attempt is to deplore recombinant proteins for use in medical sciences.

Keywords: maggot therapy, netrin-A, RACE, RAGE, lucilia sericata

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1744 Carbon Based Classification of Aquaporin Proteins: A New Proposal

Authors: Parul Johri, Mala Trivedi

Abstract:

Major Intrinsic proteins (MIPs), actively involved in the passive transport of small polar molecules across the membranes of almost all living organisms. MIPs that specifically transport water molecules are named aquaporins (AQPs). The permeability of membranes is actively controlled by the regulation of the amount of different MIPs present but also in some cases by phosphorylation and dephosphorylation of the channel. Based on sequence similarity, MIPs have been classified into many categories. All of the proteins are made up of the 20 amino acids, the only difference is there in their orientations. Again all the 20 amino acids are made up of the basic five elements namely: carbon, hydrogen, oxygen, sulphur and nitrogen. These elements are responsible for giving the amino acids the properties of hydrophilicity/hydrophobicity which play an important role in protein interactions. The hydrophobic amino acids characteristically have greater number of carbon atoms as carbon is the main element which contributes to hydrophobic interactions in proteins. It is observed that the carbon level of proteins in different species is different. In the present work, we have taken a sample set of 150 aquaporins proteins from Uniprot database and a dynamic programming code was written to calculate the carbon percentage for each sequence. This carbon percentage was further used to barcode the aqauporins of animals and plants. The protein taken from Oryza sativa, Zea mays and Arabidopsis thaliana preferred to have carbon percentage of 31.8 to 35, whereas on the other hand sequences taken from Mus musculus, Saccharomyces cerevisiae, Homo sapiens, Bos Taurus, and Rattus norvegicus preferred to have carbon percentage of 31 to 33.7. This clearly demarks the carbon range in the aquaporin proteins from plant and animal origin. Hence the atom level analysis of protein sequences can provide us with better results as compared to the residue level comparison.

Keywords: aquaporins, carbon, dynamic prgramming, MIPs

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1743 Nutritional Quality of Partially Processed Chicken Meat Products from Egyptian and Saudi Arabia Markets

Authors: Ali Meawad Ahmad, Hosny A. Abdelrahman

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Chicken meat is a good source of protein of high biological value which contains most of essential amino-acids with high proportion of unsaturated fatty acids and low cholesterol level. Besides, it contain many vitamins as well as minerals which are important for the human body. Therefore, a total of 150 frozen chicken meat product samples, 800g each within their shelf-life, were randomly collected from commercial markets from Egypt (75 samples) and Saudi Arabian (75 samples) for chemical evaluation. The mean values of fat% in the examined samples of Egyptian and Saudi markets were 16.0% and 4.6% for chicken burger; 15.0% and 11% for nuggets and 11% and 11% for strips respectively. The mean values of moisture % in the examined samples of Egyptian and Saudi markets were 67.0% and 81% for chicken burger; 66.0% and 78% for nuggets and 71.0% and 72% for strips respectively. The mean values of protein % in the examined samples of Egyptian and Saudi markets were 15% and 17% for chicken burger; 16% and 16% for nuggets and 16% and 17% for strips respectively. The obtained results were compared with the Egyptian slandered and suggestions for improving the chemical quality of chicken products were given.

Keywords: chicken meat, nutrition, Egypt, markets

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1742 LGR5 and Downstream Intracellular Signaling Proteins Play Critical Roles in the Cell Proliferation of Neuroblastoma, Meningioma and Pituitary Adenoma

Authors: Jin Hwan Cheong, Mina Hwang, Myung Hoon Han, Je Il Ryu, Young ha Oh, Seong Ho Koh, Wu Duck Won, Byung Jin Ha

Abstract:

Leucine-rich repeat-containing G-protein coupled receptor 5 (LGR5) has been reported to play critical roles in the proliferation of various cancer cells. However, the roles of LGR5 in brain tumors and the specific intracellular signaling proteins directly associated with it remain unknown. Expression of LGR5 was first measured in normal brain tissue, meningioma, and pituitary adenoma of humans. To identify the downstream signaling pathways of LGR5, siRNA-mediated knockdown of LGR5 was performed in SH-SY5Y neuroblastoma cells followed by proteomics analysis with 2-dimensional polyacrylamide gel electrophoresis (2D-PAGE). In addition, the expression of LGR5-associated proteins was evaluated in LGR5-inꠓhibited neuroblastoma cells and in human normal brain, meningioma, and pituitary adenoma tissue. Proteomics analysis showed 12 protein spots were significantly different in expression level (more than two-fold change) and subsequently identified by peptide mass fingerprinting. A protein association network was constructed from the 12 identified proteins altered by LGR5 knockdown. Direct and indirect interactions were identified among the 12 proteins. HSP 90-beta was one of the proteins whose expression was altered by LGR5 knockdown. Likewise, we observed decreased expression of proteins in the hnRNP subfamily following LGR5 knockdown. In addition, we have for the first time identified significantly higher hnRNP family expression in meningioma and pituitary adenoma compared to normal brain tissue. Taken together, LGR5 and its downstream sigꠓnaling play critical roles in neuroblastoma and brain tumors such as meningioma and pituitary adenoma.

Keywords: LGR5, neuroblastoma, meningioma, pituitary adenoma, hnRNP

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1741 Nutritional Value Determination of Different Varieties of Oats and Barley Using Near-Infrared Spectroscopy Method for the Horses Nutrition

Authors: V. Viliene, V. Sasyte, A. Raceviciute-Stupeliene, R. Gruzauskas

Abstract:

In horse nutrition, the most suitable cereal for their rations composition could be defined as oats and barley. Oats have high nutritive value because it provides more protein, fiber, iron and zinc than other whole grains, has good taste, and an activity of stimulating metabolic changes in the body. Another cereal – barley is very similar to oats as a feed except for some characteristics that affect how it is used; however, barley is lower in fiber than oats and is classified as a "heavy" feed. The value of oats and barley grain, first of all is dependent on its composition. Near-infrared spectroscopy (NIRS) has long been considered and used as a significant method in component and quality analysis and as an emerging technology for authenticity applications for cereal quality control. This paper presents the chemical and amino acid composition of different varieties of barley and oats, also digestible energy of different cereals for horses. Ten different spring barley (n = 5) and oats (n = 5) varieties, grown in one location in Lithuania, were assayed for their chemical composition (dry matter, crude protein, crude fat, crude ash, crude fiber, starch) and amino acids content, digestible amino acids and amino acids digestibility. Also, the grains digestible energy for horses was calculated. The oats and barley samples reflectance spectra were measured by means of NIRS using Foss-Tecator DS2500 equipment. The chemical components: fat, crude protein, starch and fiber differed statistically (P<0.05) between the oats and barley varieties. The highest total amino acid content between oats was determined in variety Flamingsprofi (4.56 g/kg) and the lowest – variety Circle (3.57 g/kg), and between barley - respectively in varieties Publican (3.50 g/kg) and Sebastian (3.11 g/kg). The different varieties of oats digestible amino acid content varied from 3.11 g/kg to 4.07 g/kg; barley different varieties varied from 2.59 g/kg to 2.94 g/kg. The average amino acids digestibility of oats varied from 74.4% (Liz) to 95.6% (Fen) and in barley - from 75.8 % (Tre) to 89.6% (Fen). The amount of digestible energy in the analyzed varieties of oats and barley was an average compound 13.74 MJ/kg DM and 14.85 MJ/kg DM, respectively. An analysis of the results showed that different varieties of oats compared with barley are preferable for horse nutrition according to the crude fat, crude fiber, ash and separate amino acids content, but the analyzed barley varieties dominated the higher amounts of crude protein, the digestible Liz amount and higher DE content, and thus, could be recommended for making feed formulation for horses combining oats and barley, taking into account the chemical composition of using cereal varieties.

Keywords: barley, digestive energy, horses, nutritional value, oats

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1740 Quality Rabbit Skin Gelatin with Acetic Acid Extract

Authors: Wehandaka Pancapalaga

Abstract:

This study aimed to analyze the water content, yield, fat content, protein content, viscosity, gel strength, pH, melting and organoleptic rabbit skin gelatin with acetic acid extraction levels are different. The materials used in this study were Rex rabbit skin male. Treatments that P1 = the extraction of acetic acid 2% (v / v); P2 = the extraction of acetic acid 3% (v / v); P3 = the extraction of acetic acid 4 % (v / v). P5 = the extraction of acetic acid 5% (v / v). The results showed that the greater the concentration of acetic acid as the extraction of rabbit skin can reduce the water content and fat content of rabbit skin gelatin but increase the protein content, viscosity, pH, gel strength, yield and melting point rabbit skin gelatin. texture, color and smell of gelatin rabbits there were no differences with cow skin gelatin. The results showed that the quality of rabbit skin gelatin accordance Indonesian National Standard (SNI). Conclusion 5% acetic acid extraction produces the best quality gelatin.

Keywords: gelatin, skin rabbit, acetic acid extraction, quality

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1739 Cumulus-Oocyte Complexes and Follicular Fluid Proteins of Pig during Folliculogenesis

Authors: Panomporn Wisuthseriwong, Hatairuk Tungkasen, Siyaporn Namsongsan, Chanikarn Srinark, Mayuva Youngsabanant-Areekijseree

Abstract:

The objective of the present study was to evaluate the morphology of porcine cumulus-oocyte complexes (pCOCs) and follicular fluid during follicular development. The samples were obtained from local slaughterhouses in Nakorn Pathom Province, Thailand. Pigs were classified as either in the follicular phase or luteal phase. Porcine follicles (n = 3,510) were categorized as small (1-3 mm in diameters; n=2,910), medium (4-6 mm in diameters; n=530) and large (7-8 mm in diameters; n=70). Then pCOCs and follicular fluid were collected. Finally, we found that the oocytes can be categorized into intact cumulus cells layer oocyte, multi-cumulus cells layer oocyte, partial cumulus cells layer oocyte, completely denuded oocyte and degenerated oocyte. They showed high percentage of intact and multi-cumulus cells layer oocytes from small follicles (54.68%) medium follicles (69.06%) and large follicles (68.57%), which have high potential to develop into matured oocytes in vitro. Protein composition of the follicular fluid was separated by SDS-PAGE technique. The result shows that the protein molecular weight in the small and medium follicles are 23, 50, 66, 75, 92, 100, 132, 163, 225 and >225 kDa. Meanwhile, protein molecular weight in large follicles are 12, 16, 23, 50, 66, 75, 92, 100, 132, 163, 225 and >225 kDa. All proteins play an important role in promotion and regulation on development, maturation of oocytes and regulation of ovulation. We conclude that the results of discovery can be used porcine secretion proteins for supplement in IVM/IVF technology. Acknowledgements: The project was funded by a grant from Silpakorn University Research and Development Institute (SURDI) and Faculty of Science, Silpakorn University, Thailand.

Keywords: porcine follicles, porcine oocyte, follicular fluid, SDS-PAGE

Procedia PDF Downloads 237
1738 Caffeic Acid Methyl and Ethyl Esters Exhibit Beneficial Effect on Glucose and Lipid Metabolism in Cultured Murine Insulin-Sensitive Cells

Authors: Hoda M. Eid, Abir Nachar, Farah Thong, Gary Sweeney, Pierre S. Haddad

Abstract:

Caffeic acid methyl ester (CAME) and caffeic ethyl esters (CAEE) were previously reported to potently stimulate glucose uptake in cultured C2C12 skeletal muscle cells via insulin-independent mechanisms involving the activation of adenosine monophosphate-activated protein kinase (AMPK). In the present study, we investigated the effect of the two compounds on the translocation of glucose transporter GLUT4 in L6 skeletal muscle cells. The cells were treated with the optimum non-toxic concentration (50 µM) of either CAME or CAEE for 18 h. Levels of GLUT4myc at the cell surface were measured by O-phenylenediamine dihydrochloride (OPD) assay. The effects of CAME and CAEE on GLUT1 and GLUT4 protein content were also measured by western immunoblot. Our results show that CAME and CAEE significantly increased glucose uptake, GLUT4 translocation and GLUT4 protein content. Furthermore, the effect of the two CA esters on two insulin-sensitive cell lines: H4IIE rat hepatoma and 3T3-L1 adipocytes were investigated. CAME and CAEE reduced the enzymatic activity of the key hepatic gluconeogenic enzyme glucose-6-phosphatase in a concentration-dependent manner. In addition, they exerted a concentration-dependent antiadipogenic effect on 3T3-L1 cells. Mitotic clonal expansion (MCE), a prerequisite for adipocytes differentiation was also concentration-dependently inhibited. The two compounds abrogated lipid droplet accumulation, blocked MCE and maintained cells in fibroblast-like state when applied at the maximum non-toxic concentration (100 µM). In addition, the expression of the early key adipogenic transcription factors CCAAT enhancer-binding protein beta (C/EBP-β) and the master regulator of adipogenesis peroxisome-proliferator-activated receptor gamma (PPAR-γ) were inhibited. We, therefore, conclude that CAME and CAEE exert pleiotropic benefits in several insulin-sensitive cell lines through insulin-independent mechanisms involving AMPK, hence they may treat obesity, diabetes and other metabolic diseases.

Keywords: type 2 diabetes mellitus, insulin resistance, GLUT4, Akt, AMPK.

Procedia PDF Downloads 285
1737 A Structure-Switching Electrochemical Aptasensor for Rapid, Reagentless and Single-Step, Nanomolar Detection of C-Reactive Protein

Authors: William L. Whitehouse, Louisa H. Y. Lo, Andrew B. Kinghorn, Simon C. C. Shiu, Julian. A. Tanner

Abstract:

C-reactive protein (CRP) is an acute-phase reactant and sensitive indicator for sepsis and other life-threatening pathologies, including systemic inflammatory response syndrome (SIRS). Currently, clinical turn-around times for established CRP detection methods take between 30 minutes to hours or even days from centralized laboratories. Here, we report the development of an electrochemical biosensor using redox probe-tagged DNA aptamers functionalized onto cheap, commercially available screen-printed electrodes. Binding-induced conformational switching of the CRP-targeting aptamer induces a specific and selective signal-ON event, which enables single-step and reagentless detection of CRP in as little as 1 minute. The aptasensor dynamic range spans 5-1000nM (R=0.97) or 5-500nM (R=0.99) in 50% diluted human serum, with a LOD of 3nM, corresponding to 2-orders of magnitude sensitivity under the clinically relevant cut-off for CRP. The sensor is stable for up to one week and can be reused numerous times, as judged from repeated real-time dosing and dose-response assays. By decoupling binding events from the signal induction mechanism, structure-switching electrochemical aptamer-based sensors (SS-EABs) provide considerable advantages over their adsorption-based counterparts. Our work expands on the retinue of such sensors reported in the literature and is the first instance of an SS-EAB for reagentless CRP detection. We hope this study can inspire further investigations into the suitability of SS-EABs for diagnostics, which will aid translational R&D toward fully realized devices aimed at point-of-care applications or for use more broadly by the public.

Keywords: structure-switching, C-reactive protein, electrochemical, biosensor, aptasensor.

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1736 The Effect of Fermented Organic Feed into Nutritive Contents of Kampong Chicken Meat

Authors: Wahyu Widodo, Imbang Dwi Rahayu, Adi Sutanto

Abstract:

The purpose of this research was to analyze the effect of the fermented organic feed to dry matter, ash, organic matter, protein, fat and crude fiber contents of kampong chicken meat. The research had conducted at January until June, 2016. One hundreds chickens were used in this research. Experimental method and completely randomized design were used to support this research. We had 4 treatment namely P0: organic feed without fermentation, P1: Organic feed with fermented rice bran, P2: Organic feed with fermented corn, P3: Organic feed with fermented rice bran and corn with 5 replication. The conclusion was the treatment had not a significant effect in the dry matter, ash, organic matter and protein contents of chicken meat. On the other hand, it had a significant effect in the fat and crude fiber contents of chicken meat.

Keywords: corn, fermented organic feed, nutritive contents, rice bran

Procedia PDF Downloads 294
1735 The Effect of Acute Aerobic Exercise after Consumption of Four Different Diets on Serum Levels Irisin, Insulin and Glucose in Overweight Men

Authors: Majid Mardaniyan Ghahfarokhi, Abdolhamid Habibi, Majid Mohammad Shahi

Abstract:

The combination of exercise and diet as the most important strategy to reduce weight and control obesity-related factors, including Irisin, Insulin, and Glucose was raised. The aim of this study was to investigate the effect of aerobic exercise combined with four different diets on serum levels of Irisin, Insulin, and Glucose in overweight men. Methods: In this quasi-experimental study, 8 overweight men (BMI 29.23±0.47) with average age of (23±1.6) voluntarily participated in 4 sessions by one-week interval. The study was done in exercise physiology lab. In each session, subjects performed a 30 minutes treadmill test with 60-70% of maximum heart rate, after consuming a high carbohydrate, high-fat, high-protein and normal diet. For biochemical measurement, three blood samples were taken in fasting state, two hours after meals and after exercise Results: Statistical analysis of data showed that the serum levels of Irisin after consumption all four diets had been reduced which this reduce as a result of high-fat diet that were significantly (p ≤ 0/038). Serum concentration of Insulin and Glucose increased after consuming four diets. However, increase in serum Insulin and Glucose was significant only after consuming high-carbohydrate diet (Respectively p ≤ 0/001, p ≤ 0/042). In addition, during exercise after consuming all four regular diet, high carbohydrate, high-protein and high-fat, Irisin significant increased significantly (Respectively p ≤ 0/021, p ≤ 0/049, p ≤ 0/001, P ≤ 0/003), Insulin decreased significantly (Respectively p ≤ 0/002, p ≤ 0/001, p ≤ 0/001, p ≤ 0/002) and Glucose were significantly reduced (Respectively p ≤ 0/001, p ≤ 0/001, P ≤ 0/001, p ≤ 0/002). After aerobic activity following the consumption of a high protein diet the highest increase in irisin levels, and after aerobic exercise following consumption of high carbohydrate diet the greatest decrease in insulin and glucose levels were observed. Conclusion: It seems that diet alone and exercises following different consumption diets can have a significant effect on Irisin, Insulin, and Glucose serum levels in overweight young men.

Keywords: acute aerobic exercise, diet, irisin, overweight

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1734 Characterization of (GRAS37) Gibberellin Acid Insensitive (GAI), Repressor (RGA), and Scarecrow (SCR) Gene by Using Bioinformatics Tools

Authors: Yusra Tariq

Abstract:

The Grass 37 gene is presently known in tomatoes, which are the source of healthy substances such as ascorbic acid, polyphenols, carotenoids and nutrients. It has a significant impact on the growth and development of humans. The GRASS 37 gene is a plant Transcription factor group assuming significant parts in various reactions of different Abiotic stresses such as (drought, salinity, thermal stresses, temperature, and bright waves) which could highly affect the growth. Tomatoes are very sensitive to temperature, and their growth or production occurs optimally in a temperature range from 21 C to 29.5 C during the daytime and from 18.5 C to 21 C during the night. This protein acts as a positive regulator of salt stress response and abscisic acid signaling. This study summarizes the structure characterized by molecular formula and protein-binding domains by different bioinformatics tools such as Expasy translate tool, Expasy Portparam, Swiss Prot and Inter Pro Scan, Clustal W tool regulatory procedure of GRASS gene components, also their reactions to both biotic and Abiotic stresses.

Keywords: GRAS37, gene, bioinformatics, tool

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1733 Manipulating The PAAR Proteins of Acinetobacter Baumannii

Authors: Irene Alevizos, Jessica Lewis, Marina Harper, John Boyce

Abstract:

Acinetobacter baumannii causes a range of severe nosocomial-acquired infections, and many strains are multi-drug resistant. A. baumannii possesses survival mechanisms allowing it to thrive in competitive polymicrobial environments, including a Type VI Secretion System (T6SS) that injects effector proteins into other bacteria to give a competitive advantage. The effects of T6SS firing are broad and depend entirely on the effector that is delivered. Effects can include toxicity against prokaryotic or eukaryotic cells and the acquisition of essential nutrients. The T6SS of some species can deliver ‘specialised effectors’ that are fused directly to T6SS components, such as PAAR proteins. PAAR proteins are predicted to form the piercing tip of the T6SS and are essential for T6SS function. Although no specialised effectors have been identified in A. baumannii, many strains encode multiple PAAR proteins. Analysis of PAAR proteins across the species identified 12 families of PAAR proteins with distinct C-terminal extensions. A. baumannii AB307-0294 encodes two PAAR proteins, one of which has a C-terminal extension. Mutation of one or both of the PAAR-encoding genes in this strain showed that expression of either PAAR protein was sufficient for T6SS function. We employed a heterologous expression approach and determined that PAAR proteins from different A. baumannii strains, as well as the closely related A. baylyi species, could complement the A. baumannii ∆paar mutant and restore T6SS function. Furthermore, we showed that PAAR fusions could be used to deliver artificially cloned protein fragments by generating Histidine- and Streptavidin- tagged PAAR specialised effectors, which restored T6SS activity. This provides evidence that the fusion of protein fragments onto PAAR proteins in A. baumannii is compatible with a functional T6SS. Successful delivery by this mechanism extends the scope of what the T6SS can deliver, including user designed proteins.

Keywords: A. baumannii, effectors, PAAR, T6SS

Procedia PDF Downloads 70
1732 Incorporation of Noncanonical Amino Acids into Hard-to-Express Antibody Fragments: Expression and Characterization

Authors: Hana Hanaee-Ahvaz, Monika Cserjan-Puschmann, Christopher Tauer, Gerald Striedner

Abstract:

Incorporation of noncanonical amino acids (ncAA) into proteins has become an interesting topic as proteins featured with ncAAs offer a wide range of different applications. Nowadays, technologies and systems exist that allow for the site-specific introduction of ncAAs in vivo, but the efficient production of proteins modified this way is still a big challenge. This is especially true for 'hard-to-express' proteins where low yields are encountered even with the native sequence. In this study, site-specific incorporation of azido-ethoxy-carbonyl-Lysin (azk) into an anti-tumor-necrosis-factor-α-Fab (FTN2) was investigated. According to well-established parameters, possible site positions for ncAA incorporation were determined, and corresponding FTN2 genes were constructed. Each of the modified FTN2 variants has one amber codon for azk incorporated either in its heavy or light chain. The expression level for all variants produced was determined by ELISA, and all azk variants could be produced with a satisfactory yield in the range of 50-70% of the original FTN2 variant. In terms of expression yield, neither the azk incorporation position nor the subunit modified (heavy or light chain) had a significant effect. We confirmed correct protein processing and azk incorporation by mass spectrometry analysis, and antigen-antibody interaction was determined by surface plasmon resonance analysis. The next step is to characterize the effect of azk incorporation on protein stability and aggregation tendency via differential scanning calorimetry and light scattering, respectively. In summary, the incorporation of ncAA into our Fab candidate FTN2 worked better than expected. The quantities produced allowed a detailed characterization of the variants in terms of their properties, and we can now turn our attention to potential applications. By using click chemistry, we can equip the Fabs with additional functionalities and make them suitable for a wide range of applications. We will now use this option in a first approach and develop an assay that will allow us to follow the degradation of the recombinant target protein in vivo. Special focus will be laid on the proteolytic activity in the periplasm and how it is influenced by cultivation/induction conditions.

Keywords: degradation, FTN2, hard-to-express protein, non-canonical amino acids

Procedia PDF Downloads 207
1731 Computer Aided Screening of Secreted Frizzled-Related Protein 4 (SFRP4): A Potential Control for Diabetes Mellitus

Authors: Shazia Anwer Bukhari, Waseem Akhtar Shamshari, Mahmood-Ur-Rahman, Muhammad Zia-Ul-Haq, Hawa Z. E. Jaafar

Abstract:

Diabetes mellitus is a life threatening disease and scientists are doing their best to find a cost effective and permanent treatment of this malady. The recent trend is to control the disease by target base inhibiting of enzymes or proteins. Secreted frizzled-related protein 4 (SFRP4) is found to cause five times more risk of diabetes when expressed above average levels. This study was therefore designed to analyze the SFRP4 and to find its potential inhibitors. SFRP4 was analyzed by bio-informatics tools of sequence tool and structure tool. A total of three potential inhibitors of SFRP4 were found, namely cyclothiazide, clopamide and perindopril. These inhibitors showed significant interactions with SFRP4 as compared to other inhibitors as well as control (acetohexamide). The findings suggest the possible treatment of diabetes mellitus type 2 by inhibiting the SFRP4 using the inhibitors cyclothiazide, clopamide and perindopril.

Keywords: bioscreening, clopamide, cyclothiazide, diabetes mellitus, perindopril, SFRP4

Procedia PDF Downloads 428
1730 Exploring the Biochemical and Therapeutic Properties of Aged Garlic

Authors: Farhan Saeed

Abstract:

The core objective of this work is to explicate the biochemical and therapeutic properties of aged garlic. For this purpose, two varieties of garlic were obtained from Ayub Agricultural Research Institute (AARI) Faisalabad-Pakistan. Additionally, fresh garlic was converted into aged garlic via fermentation method in the incubator at 70 to 80 % humidity level and 60C0 temperature for one month. Similarly, biochemical and antioxidant properties of fresh and aged garlic were also elucidated. Mean values showed that moisture content was decreased, whereas crude fat, crude protein, crude fiber, crude ash and total carbohydrates were enhanced after fermentation. Additionally, crude protein of fresh and aged garlic was 7.57±0.16 and 5.52±0.12%, respectively, whilst 9.68±0.41 and 8.78±0.29%, respectively, after the fermentation process. In addition, NFE contents were also enhanced up to 39% after the fermentation method. Moreover, Zn, S, Al, K, Fe, Na, Mg, and Cu contents were also increased. Furthermore, Total phenolic contents (TPC) of fresh and aged garlic were 2498.70 & 2188.50mg GAE/kg whilst 3008.59, & 2591.81mg GAE/kg for aged garlic. In conclusion, aged garlic explicated the better biochemical properties, mineral profile and antioxidant properties as compared to fresh garlic.

Keywords: aged garlic, nutritional values, bioactive properties, fermentation

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1729 The Effect of Elapsed Time on the Cardiac Troponin-T Degradation and Its Utility as a Time Since Death Marker in Cases of Death Due to Burn

Authors: Sachil Kumar, Anoop K.Verma, Uma Shankar Singh

Abstract:

It’s extremely important to study postmortem interval in different causes of death since it assists in a great way in making an opinion on the exact cause of death following such incident often times. With diligent knowledge of the interval one could really say as an expert that the cause of death is not feigned hence there is a great need in evaluating such death to have been at the CRIME SCENE before performing an autopsy on such body. The approach described here is based on analyzing the degradation or proteolysis of a cardiac protein in cases of deaths due to burn as a marker of time since death. Cardiac tissue samples were collected from (n=6) medico-legal autopsies, (Department of Forensic Medicine and Toxicology), King George’s Medical University, Lucknow India, after informed consent from the relatives and studied post-mortem degradation by incubation of the cardiac tissue at room temperature (20±2 OC) for different time periods (~7.30, 18.20, 30.30, 41.20, 41.40, 54.30, 65.20, and 88.40 Hours). The cases included were the subjects of burn without any prior history of disease who died in the hospital and their exact time of death was known. The analysis involved extraction of the protein, separation by denaturing gel electrophoresis (SDS-PAGE) and visualization by Western blot using cTnT specific monoclonal antibodies. The area of the bands within a lane was quantified by scanning and digitizing the image using Gel Doc. As time postmortem progresses the intact cTnT band degrades to fragments that are easily detected by the monoclonal antibodies. A decreasing trend in the level of cTnT (% of intact) was found as the PM hours increased. A significant difference was observed between <15 h and other PM hours (p<0.01). Significant difference in cTnT level (% of intact) was also observed between 16-25 h and 56-65 h & >75 h (p<0.01). Western blot data clearly showed the intact protein at 42 kDa, three major (28 kDa, 30kDa, 10kDa) fragments, three additional minor fragments (12 kDa, 14kDa, and 15 kDa) and formation of low molecular weight fragments. Overall, both PMI and cardiac tissue of burned corpse had a statistically significant effect where the greatest amount of protein breakdown was observed within the first 41.40 Hrs and after it intact protein slowly disappears. If the percent intact cTnT is calculated from the total area integrated within a Western blot lane, then the percent intact cTnT shows a pseudo-first order relationship when plotted against the time postmortem. A strong significant positive correlation was found between cTnT and PM hours (r=0.87, p=0.0001). The regression analysis showed a good variability explained (R2=0.768) The post-mortem Troponin-T fragmentation observed in this study reveals a sequential, time-dependent process with the potential for use as a predictor of PMI in cases of burning.

Keywords: burn, degradation, postmortem interval, troponin-T

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1728 Nutritive Advantage of Mealworm (Tenebrio molitor) in the Diet of White Shrimp (Litopenaeus vannamei)

Authors: Tae-ho Chung, Chul Park, Gi-wook Shin, Joo-min Kim, Seong-hyun Kim, Namjung Kim

Abstract:

Mealworm (Tenebrio molitor) was evaluated to investigate the effect of partial or total replacement of fish meal in diets for white shrimp, Litopenaeus vannamei. Experimental groups of shrimp with average initial body weight (2.43 ± 0.54 g) were fed each with 4 isonitrogeneous (38% crude protein) diets formulated to include 0, 25, 50 and 100% (diets 1 to 4, respectively) of fish meal substituted with mealworm. After eight weeks of feeding trials, shrimp fed with diet 3 and 4 revealed the highest values for live weight gain(8.01 ± 2.51 and 7.93 ± 1.12), specific growth rates (2.70 ± 1.12 and 2.59 ± 0.51) as well as better feed conversion ratio (2.69 ± 0.09 and 2.72 ± 0.19) compared to the control group with statistically significant manner (p<0.05). Survival range was 98% in all the treatments. An increase in weight gain and other growth associated parameters was observed with higher replacement. These results clearly indicate that 50% and 100% of fish meal protein in shrimp diet can be replaced by mealworm not only without any adverse effect but also the effect of promoting growth performance.

Keywords: mealworm, Litopenaeus vannamei, Tenebrio molitor, white shrimp

Procedia PDF Downloads 446
1727 Molecular Implication of Interaction of Human Enteric Pathogens with Phylloplane of Tomato

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

Abstract:

Cultivation and consumption of organically grown fruits and vegetables have increased by several folds. However, the presence of Human Enteric Pathogens on the surface of organically grown vegetables causing Gastro-intestinal diseases, are most likely due to contaminated water and fecal matter of farm animals. Human Enteric Pathogens are adapted to colonize the human gut, and also colonize plant surface. Microbes on plant surface communicate with each other to establish quorum sensing. The cross talk study is important because the enteric pathogens on phylloplane have been reported to mask the beneficial resident bacteria of plant. In the present study, HEPs and bacterial colonizers were identified using 16s rRNA sequencing. Microbial colonization patterns after interaction between Human Enteric Pathogens and natural bacterial residents on tomato phylloplane was studied. Tomato plants raised under aseptic conditions were inoculated with a mixture of Serratia fonticola and Klebsiella pneumoniae. The molecules involved in cross-talk between Human Enteric Pathogens and regular bacterial colonizers were isolated and identified using molecular techniques and HPLC. The colonization pattern was studied by leaf imprint method after 48 hours of incubation. The associated protein-protein interaction in the host cytoplasm was studied by use of crosslinkers. From treated leaves the crosstalk molecules and interaction proteins were separated on 1D SDS-PAGE and analyzed by MALDI-TOF-TOF analysis. The study is critical in understanding the molecular aspects of HEP’s adaption to phylloplane. The study revealed human enteric pathogens aggressively interact among themselves and resident bacteria. HEPs induced establishment of a signaling cascade through protein-protein interaction in the host cytoplasm. The study revealed that the adaptation of Human Enteric Pathogens on phylloplane of Solanum lycopersicum involves the establishment of complex molecular interaction between the microbe and the host including microbe-microbe interaction leading to an establishment of quorum sensing. The outcome will help in minimizing the HEP load on fresh farm produce, thereby curtailing incidences of food-borne diseases.

Keywords: crosslinkers, human enteric pathogens (HEPs), phylloplane, quorum sensing

Procedia PDF Downloads 251