Search results for: tigh junction proteins

Authors: Arun Prasad, Abhishek Tiwari, Rekha Jaiswal, Vivek Chaudhary

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Background: Robotic Roux en Y gastric bypass is being done for some time but is technically difficult, requiring operating in both the sub diaphragmatic and infracolic compartments of the abdomen. This can mean a dual docking of the robot or a hybrid partial laparoscopic and partial robotic surgery. The Mini /One anastomosis /omega loop gastric bypass (MGB) has the advantage of having all dissection and anastomosis in the supracolic compartment and is therefore suitable technically for robotic surgery. Methods: We have done 208 robotic mini gastric bypass surgeries. The robot is docked above the head of the patient in the midline. Camera port is placed supra umbilically. Two ports are placed on the left side of the patient and one port on the right side of the patient. An assistant port is placed between the camera port and right sided robotic port for use of stapler. Distal stomach is stapled from the lesser curve followed by a vertical sleeve upwards leading to a long sleeve pouch. Jejunum is taken at 200 cm from the duodenojejunal junction and brought up to do a side to side gastrojejunostomy. Results: All patients had a successful robotic procedure. Mean time taken was 85 minutes. There were major intraoperative or post operative complications. No patient needed conversion or re-explorative surgery. Mean excess weight loss over a period of 2 year was about 75%. There was no mortality. Patient satisfaction score was high and was attributed to the good weight loss and minimal dietary modifications that were needed after the procedure. Long term side effects were anemia and bile reflux in a small number of patients. Conclusions: MGB / OAGB is gaining worldwide interest as a short simple procedure that has been shown to very effective and safe bariatric surgery. The purpose of this study was to report on the safety and efficacy of robotic surgery for this procedure. This is the first report of totally robotic mini gastric bypass.

Keywords: MGB, mini gastric bypass, OAGB, robotic bariatric surgery

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Authors: Jovana Petrović, Ivana Lončarević, Vesna Tumbas Šaponjac, Biljana Pajin, Danica Zarić

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Pomace, a by-product from fruit processing industry is the potential source of valuable bioactive. Cookies are popular, ready to eat and low price foods; therefore, enrichment of these products is of great importance. In this work, bioactive compounds extracted from cherry pomace, encapsulated in soy and whey proteins, have been incorporated in cookies, replacing 10 (SP10 and WP10) and 15% of wheat flour (SP15 and WP15). Cookie geometry (diameter (D), thickness (T) and spread ratio (D/T)), cookie weight, cookie hardness and cookie surface colour were measured. Sensory characteristics are also examined. The results show that encapsulated cherry pomace bioactives have positively influenced the cookie mass. Diameter, redness (a* value) and cookie hardness increased. Sensory evaluation of cookies, revealed that up to 15% substitution of wheat flour with WP encapsulate produced acceptable cookies similar to the control (100% wheat flour) cookies.

Keywords: cherry pomace, polyphenols, microencapsulation, cookies, physical characteristics

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Authors: Dileep Kumar, Janhavi Ramchandra Rao Kumar, Rao

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COVID-19 has ravaged the globe, and it is spreading its Spectre day by day. In the absence of established drugs, this disease has created havoc. Some of the infected persons are symptomatic or asymptomatic. The respiratory system, cardiac system, digestive system, etc. in human beings are affected by this virus. In our present investigation, we have undertaken a study of the Indian Ayurvedic herb, Ocimum sanctum against SARS-CoV-2 using molecular docking and dynamics studies. The docking analysis was performed on the Glide module of Schrödinger suite on two different proteins from SARS-CoV-2 viz. NSP15 Endoribonuclease and spike receptor-binding domain. MM-GBSA based binding free energy calculations also suggest the most favorable binding affinities of carvacrol, β elemene, and β caryophyllene with binding energies of −61.61, 58.23, and −54.19 Kcal/mol respectively with spike receptor-binding domain and NSP15 Endoribonuclease. It rekindles our hope for the design and development of new drug candidates for the treatment of COVID19.

Keywords: molecular docking, COVID-19, ocimum sanctum, binding energy

Procedia PDF Downloads 187

Authors: Saiada Fuadi Fancy, Fahim Ahmed, Shofiq Ahmed, Raquib Ahsan

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The worldwide use of reinforced concrete construction stems from the wide availability of reinforcing steel as well as concrete ingredients. However, concrete construction requires a certain level of technology, expertise, and workmanship, particularly, in the field during construction. As a supporting technology for a concrete column or wall construction, kicker is cast as part of the slab or foundation to provide a convenient starting point for a wall or column ensuring integrity at this important junction. For that reason, a comprehensive study was carried out here to investigate the behavior of reinforced concrete frame with different kicker parameters. To achieve this objective, six half-scale specimens of portal reinforced concrete frame with kickers and one portal frame without kicker were constructed according to common practice in the industry and subjected to cyclic incremental horizontal loading with sustained gravity load. In this study, the experimental data, obtained in four deflections controlled cycle, were used to evaluate the behavior of kickers. Load-displacement characteristics were obtained; maximum loads and deflections were measured and assessed. Finally, the test results of frames constructed with three different types of kicker thickness were compared with the kickerless frame. Similar crack patterns were observed for all the specimens. From this investigation, specimens with kicker thickness 3″ were shown better results than specimens with kicker thickness 1.5″, which was specified by maximum load, stiffness, initiation of first crack and residual displacement. Despite of better performance, it could not be firmly concluded that 4.5″ kicker thickness is the most appropriate one. Because, during the test of that specimen, separation of dial gauge was needed. Finally, comparing with kickerless specimen, it was observed that performance of kickerless specimen was relatively better than kicker specimens.

Keywords: crack, cyclic, kicker, load-displacement

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Authors: M. Markova, E. Filova, O. Kaplan, R. Matejka, L. Bacakova

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The porcine pericardium is used as a material for cardiac and aortic valves substitutes. Current biological aortic heart valve prosthesis have a limited lifetime period because they undergo degeneration. In order to make them more biocompatible and prolong their lifetime it is necessary to reseed the decellularized prostheses with endothelial cells and with valve interstitial cells. The endothelialization of the prosthesis-surface may be supported by suitable chemical surface modification of the prosthesis. The aim of this study is to prepare bioactive fibrin layers which would both support endothelialization of porcine pericardium and enhance differentiation and maturation of the endothelial cells seeded. As a material for surface adjustments we used layers of fibrin with/without heparin and some of them with adsorbed or chemically bound FGF2, VEGF or their combination. Fibrin assemblies were prepared in 24-well cell culture plate and were seeded with HSVEC (Human Saphenous Vein Endothelial Cells) at a density of 20,000 cells per well in EGM-2 medium with 0.5% FS and without heparin, without FGF2 and without VEGF; medium was supplemented with aprotinin (200 U/mL). As a control, surface polystyrene (PS) was used. Fibrin was also used as homogeneous impregnation of the decellularized porcine pericardium throughout the scaffolds. Morphology, density, and viability of the seeded endothelial cells were observed from micrographs after staining the samples by LIVE/DEAD cytotoxicity/viability assay kit on the days 1, 3, and 7. Endothelial cells were immunocytochemically stained for proteins involved in cell adhesion, i.e. alphaV integrin, vinculin, and VE-cadherin, markers of endothelial cells differentiation and maturation, i.e. von Willebrand factor and CD31, and for extracellular matrix proteins typically produced by endothelial cells, i.e. type IV collagen and laminin. The staining intensities were subsequently quantified using a software. HSVEC cells grew on each of the prepared surfaces better than on control surface. They reached confluency. The highest cell densities were obtained on the surface of fibrin with heparin and both grow factors used together. Intensity of alphaV integrins staining was highest on samples with remained fibrin layer, i.e. on layers with lower cell densities, i.e. on fibrin without heparin. Vinculin staining was apparent, but was rather diffuse, on fibrin with both FGF2 and VEGF and on control PS. Endothelial cells on all samples were positively stained for von Willebrand factor and CD31. VE-cadherin receptors clusters were best developed on fibrin with heparin and growth factors. Significantly stronger staining of type IV collagen was observed on fibrin with heparin and both growth factors. Endothelial cells on all samples produced laminin-1. Decellularized pericardium was homogeneously filled with fibrin structures. These fibrin-modified pericardium samples will be further seeded with cells and cultured in a bioreactor. Fibrin layers with/without heparin and with adsorbed or chemically bound FGF2, VEGF or their combination are good surfaces for endothelialization of cardiovascular prostheses or porcine pericardium based heart valves. Supported by the Ministry of Health, grants No15-29153A and 15-32497A, and the Grant Agency of the Czech Republic, project No. P108/12/G108.

Keywords: aortic valves prosthesis, FGF2, heparin, HSVEC cells, VEGF

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Authors: Arsyam Mawardi, Sony Suhandono, Azzania Fibriani, Fifi Fitriyah Masduki

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Malaria is an infectious disease caused by Plasmodium sp. This disease has a high prevalence in Indonesia, especially in Jayapura. The vaccine that is currently being developed has not been effective in overcoming malaria. This is due to the high polymorphism in the Plasmodium genome especially in areas that encode Plasmodium surface proteins. Merozoite Surface Protein 1 (MSP1) Plasmodium falciparum is a surface protein that plays a role in the invasion process in human erythrocytes through the interaction of Glycophorin A protein receptors and sialic acid in erythrocytes with Reticulocyte Binding Proteins (RBP) and Duffy Adhesion Protein (DAP) ligands in merozoites. MSP1 can be targeted to be a specific antigen and predicted epitope area which will be used for the development of diagnostic and malaria vaccine therapy. MSP1 consists of 17 blocks, each block is dimorphic, and has been marked as the K1 and MAD20 alleles. Exceptions only in block 2, because it has 3 alleles, among others K1, MAD20 and RO33. These polymorphisms cause allelic variations and implicate the severity of patients infected P. falciparum. In addition, polymorphism of MSP1 in Jayapura isolates has not been reported so it is interesting to be further identified and projected as a specific antigen. Therefore, in this study, we analyzed the allele polymorphism as well as detected the MSP1 epitope antigen candidate on block 2 P. falciparum. Clinical samples of selected malaria patients followed the consecutive sampling method, examining malaria parasites with blood preparations on glass objects observed through a microscope. Plasmodium DNA was isolated from the blood of malarial positive patients. The block 2 MSP1 gene was amplified using PCR method and cloned using the pGEM-T easy vector then transformed to TOP'10 E.coli. Positive colonies selection was performed with blue-white screening. The existence of target DNA was confirmed by PCR colonies and DNA sequencing methods. Furthermore, DNA sequence analysis was done through alignment and formation of a phylogenetic tree using MEGA 6 software and insilico analysis using IEDB software to predict epitope candidate for P. falciparum. A total of 15 patient samples have been isolated from Plasmodium DNA. PCR amplification results show the target gene size about ± 1049 bp. The results of MSP1 nucleotide alignment analysis reveal that block 2 MSP1 genes derived from the sample of malarial patients were distributed in four different allele family groups, K1 (7), MAD20 (1), RO33 (0) and MSP1_Jayapura (10) alleles. The most commonly appears of the detected allele is MSP1_Jayapura single allele. There was no significant association between sex variables, age, the density of parasitemia and alel variation (Mann Whitney, U > 0.05), while symptomatic signs have a significant difference as a trigger of detectable allele variation (U < 0.05). In this research, insilico study shows that there is a new epitope antigen candidate from the MSP1_Jayapura allele and it is predicted to be recognized by B cells with 17 amino acid lengths in the amino acid sequence 187 to 203.

Keywords: epitope candidate, insilico analysis, MSP1 P. falciparum, polymorphism

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Authors: Cristiano Giordani, Marco Diociaiuti, Cecilia Bombelli, Laura Zanetti-Polzi, Marcello Belfiore, Raoul Fioravanti, Gianfranco Macchia

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We investigated induced functional effects by evaluating Ca2+-influx in liposomes and cell viability in HT22-DIFF neurons. Only solutions rich in unstructured Prefibrillar-Oligomers (PFOs) were able, in the presence of Monosialoganglioside-GM1 (GM1), to induce Ca2+-influx and were also neurotoxic, suggesting a correlation between the two phenomena. Thus, in the presence of GM1, we investigated the protein conformation and liposome modification due to the interaction. Circular Dichroism showed that GM1 fostered the formation of β-structures and Energy Filtered-Transmission Electron Microscopy that PFOs formed “amyloid-channels” as reported for Aβ. We speculate that electrostatic forces occurring between the positive PFOs and negative GM1 drive the initial binding, while the hydrophobic profile of the flexible PFO is responsible for the subsequent pore formation. Conversely, the rigid β-structured mature/fibers (MFs) and proto-fibers (PFs) were unable to induce membrane damage and Ca2+- influx.

Keywords: amyloid proteins, neurotoxicity, lipid-rafts, GM1

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Authors: Harche Rima, Laoufi Nadia Aicha

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This work focuses on the study and modeling of the fouling phenomenon in a vertical pipe. In the first step, milk is one of the fluids obeying the phenomenon of fouling because of the denaturation of these proteins, especially lactoglobulin, which is the active element of milk, and to facilitate its use, we chose to study milk as a fouling fluid. In another step, we consider the test section of our installation as a tubular-type heat exchanger that works against the current and in a closed circuit. A simple mathematical model of Kern & Seaton, based on the kinetics of the fouling resistance, was used to evaluate the influence of the operating parameters (fluid flow velocity and exchange wall temperature) on the fouling resistance. The influence of the variation of the fouling resistance with the operating conditions on the efficiency of the heat exchanger and the importance of the dirty state exchange coefficient as an exchange quality control parameter were discussed and examined. On the other hand, an electronic scanning microscope analysis was performed on the milk deposit in order to obtain its actual image and composition, which allowed us to calculate the thickness of this deposit.

Keywords: fouling, milk, tubular heat exchanger, fouling resistance

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Authors: Priyanka Jain, Ashish K. Sharma, Esha Shukla, K. J. Mukherjee

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We propose a paradigm shift in our approach to design improved platforms for recombinant protein production, by addressing system level issues rather than the individual steps associated with recombinant protein synthesis like transcription, translation, etc. We demonstrate that by controlling and modulating the cellular stress response (CSR), which is responsible for feedback control of protein synthesis, we can generate hyper-producing strains. We did transcriptomic profiling of post-induction cultures, expressing different types of protein, to analyze the nature of this cellular stress response. We found significant down-regulation of substrate utilization, translation, and energy metabolism genes due to generation CSR inside the host cell. However, transcription profiling has also shown that many genes are up-regulated post induction and their role in modulating the CSR is unclear. We hypothesized that these up-regulated genes trigger signaling pathways, generating the CSR and concomitantly reduce the recombinant protein yield. To test this hypothesis, we knocked out the up-regulated genes, which did not have any downstream regulatees, and analyzed their impact on cellular health and recombinant protein expression. Two model proteins i.e., GFP and L-Asparaginase were chosen for this analysis. We observed a significant improvement in expression levels, with some knock-outs showing more than 7-fold higher expression compared to control. The 10 best single knock-outs were chosen to make 45 combinations of all possible double knock-outs. A further increase in expression was observed in some of these double knock- outs with GFP levels being highest in a double knock-out ΔyhbC + ΔelaA. However, for L-Asparaginase which is a secretory protein, the best results were obtained using a combination of ΔelaA+ΔcysW knock-outs. We then tested all the knock outs for their ability to enhance the expression of a 'difficult-to-express' protein. The Rubella virus E1 protein was chosen and tagged with sfGFP at the C-terminal using a linker peptide for easy online monitoring of expression of this fusion protein. Interestingly, the highest increase in Rubella-sGFP levels was obtained in the same double knock-out ΔelaA + ΔcysW (5.6 fold increase in expression yield compared to the control) which gave the highest expression for L-Asparaginase. However, for sfGFP alone, the ΔyhbC+ΔmarR knock-out gave the highest level of expression. These results indicate that there is a fair degree of commonality in the nature of the CSR generated by the induction of different proteins. Transcriptomic profiling of the double knock out showed that many genes associated with the translational machinery and energy biosynthesis did not get down-regulated post induction, unlike the control where these genes were significantly down-regulated. This confirmed our hypothesis of these genes playing an important role in the generation of the CSR and allowed us to design a strategy for making better expression hosts by simply knocking out key genes. This strategy is radically superior to the previous approach of individually up-regulating critical genes since it blocks the mounting of the CSR thus preventing the down-regulation of a very large number of genes responsible for sustaining the flux through the recombinant protein production pathway.

Keywords: cellular stress response, GFP, knock-outs, up-regulated genes

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Authors: Salmin K. Alshalmani, Nada H. Zobi, Ismaeel H. Bozakouk

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Marine organisms are potentially prolific sources of highly bio active secondary metabolites that might represent useful leads in the development of new pharmaceutical agents. The Libyan marine biodiversity including macroalgae remains partially unexplored in term of their potential bio activities. The phytochemical analysis of the alcoholic extracts of some commonly occurring seaweed Cystoseira compressa, enteromorpha intestinals, corallina, and Ulva lactuca and their evaluated for antibacterial activity by well diffusion assay were studied. Four different solvents namely water, ethanol 99 %, methanol 99 %, and methylated spirit 95 % were used for extraction. The phytochemical analysis revealed the presence of carbohydrates, steroids, tannin & phenols, saponins, proteins, and glycosides. The extracts were subjected for study of antibacterial activity. The zone of inhibition ranged between 8 to 16 mm in aqueous extract and up to 16 mm in methanol extract. The maximum activity (16 mm) was recorded from methanol extract of Ulva lactuca against Staphylococcus aureus and, minimum activity (8mm) recorded by Cystoseira compressa against S. aureus.

Keywords: macroalgae, phytochemicals, antibacterial activity, methanolic extract

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Authors: T. Büyüksırıt, H. Kuleaşan

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Natural antimicrobials are used to preserve foods that can be found in plants, animals, and microorganisms. Antimicrobial substances are natural or artificial agents that produced by microorganisms or obtained semi/total chemical synthesis are used at low concentrations to inhibit the growth of other microorganisms. Food borne pathogens and spoilage microorganisms are inactivated by the use of antagonistic microorganisms and their metabolites. Yeasts can produce toxic proteins or glycoproteins (toxins) that cause inhibition of sensitive bacteria and yeast species. Antimicrobial substance producing phenotypes belonging different yeast genus were isolated from different sources. Toxins secreted by many yeast strains inhibiting the growth of other yeast strains. These strains show antimicrobial activity, inhibiting the growth of mold and bacteria. The effect of antimicrobial agents produced by yeasts can be extremely fast, and therefore may be used in various treatment procedures. Rapid inhibition of microorganisms is possibly caused by microbial cell membrane lipopolysaccharide binding and in activation (neutralization) effect. Antimicrobial agents inhibit the target cells via different mechanisms of action.

Keywords: antimicrobial agents, yeast, toxic protein, glycoprotein

Procedia PDF Downloads 362

Authors: Wenfa Yu, Thuva Gnanasampanthan, John Finlay, Jessica Clarke, Charlotte Anderson, Tony Clare, Axel Rosenhahn

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Marine biofouling is a worldwide problem at vast economic and ecological costs. Historically it was combated with toxic coatings such as tributyltin. As those coatings being banned nowadays, finding environmental friendly antifouling solution has become an urgent topic. In this study antifouling coatings consisted of natural occurring polysaccharides hyaluronic acid (HA), alginic acid (AA), chitosan (Ch) and polyelectrolyte polyethylenimine (PEI) are constructed into polyelectrolyte multilayers (PEMs) in a Layer-by-Layer (LbL) method. LbL PEM construction is a straightforward way to assemble biomacromolecular coatings on surfaces. Advantages about PEM include ease of handling, highly diverse PEM composition, precise control over the thickness and so on. PEMs have been widely employed in medical application and there are numerous studies regarding their protein adsorption, elasticity and cell adhesive properties. With the adjustment of coating composition, termination layer charge, coating morphology and cross-linking method, it is possible to prepare low marine biofouling coatings with PEMs. In this study, using spin coating technology, PEM construction was achieved at smooth multilayers with roughness as low as 2nm rms and highly reproducible thickness around 50nm. To obtain stability in sea water, the multilayers were covalently cross-linked either thermally or chemically. The cross-linking method affected surface energy, which was reflected in water contact angle, thermal cross-linking led to hydrophobic surfaces and chemical cross-linking generated hydrophilic surfaces. The coatings were then evaluated regarding its protein resistance and biological species resistance. While the hydrophobic thermally cross-linked PEM had low resistance towards proteins, the resistance of chemically cross-linked PEM strongly depended on the PEM termination layer and the charge of the protein, opposite charge caused high adsorption and same charge low adsorption, indicating electrostatic interaction plays a crucial role in the protein adsorption processes. Ulva linza was chosen as the biological species for antifouling performance evaluation. Despite of the poor resistance towards protein adsorption, thermally cross-linked PEM showed good resistance against Ulva spores settlement, the chemically cross-linked multilayers showed poor resistance regardless of the termination layer. Marine species adhesion is a complex process, although it involves proteins as bioadhesives, protein resistance its own is not a fully indicator for its antifouling performance. The species will pre select the surface, responding to cues like surface energy, chemistry, or charge and so on. Thus making it difficult for one single factors to determine its antifouling performance. Preparing PEM coating is a comprehensive work involving choosing polyelectrolyte combination, determining termination layer and the method for cross-linking. These decisions will affect PEM properties such as surface energy, charge, which is crucial, since biofouling is a process responding to surface properties in a highly sensitive and dynamic way.

Keywords: hyaluronic acid, polyelectrolyte multilayers, protein resistance, Ulva linza zoospores

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Authors: A. T. Adetunji, F. B. Lewu, R. Mundembe

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Plants require nitrogen (N) to support desired production levels. Nitrogen is available to plants in the form of nitrate or ammonium, which are transported into the cell with the aid of various transport proteins. Ammonium transporters (AMTs) play a role in the uptake of ammonium, the form in which nitrogen is preferentially absorbed by plants. Solanum tuberosum AMT1 (StAMT1) was characterized using molecular biology and bioinformatics methods. Nucleotide database sequences were used to design AMT1-specific primers which were used to amplify the AMT1 internal regions. Nucleotide sequencing, alignment and phylogenetic analysis assigned StAMT1 to the AMT1 family. The deduced amino acid sequences showed that StAMT1 is 92%, 83% and 76% similar to Solanum lycopersicum LeAMT1.1, Lotus japonicus LjAMT1.1 and Solanum lycopersicum LeAMT1.2 respectively. StAMT1 fragments were shown to correspond to the 5th - 10th trans-membrane domains. Residue StAMT1 D15 is predicted to be essential for ammonium transport, while mutations of StAMT1 S76A may further enhance ammonium transport.

Keywords: ammonium transporter, bioinformatics, nitrogen, primers, Solanum tuberosum

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Authors: Ewelina Wisnik, Zsolt Regdon, Kinga Chmielewska, Laszlo Virag, Agnieszka Robaszkiewicz

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Apart from protecting genome, PARP1 has been documented to regulate many intracellular processes inter alia gene transcription by physically interacting with chromatin bound proteins and by their ADP-ribosylation. Our recent findings indicate that expression of PARP1 decreases during the differentiation of human CD34+ hematopoietic stem cells to monocytes as a consequence of differentiation-associated cell growth arrest and formation of E2F4-RBL2-HDAC1-SWI/SNF repressive complex at the promoter of this gene. Since the RBL2 complexes repress genes in a E2F-dependent manner and are widespread in the genome in G0 arrested cells, we asked (a) if RBL2 directly contributes to defining monocyte phenotype and function by targeting gene promoters and (b) if RBL2 controls gene transcription indirectly by repressing PARP1. For identification of genes controlled by RBL2 and/or PARP1,we used primer libraries for surface receptors and TLR signaling mediators, genes were silenced by siRNA or shRNA, analysis of gene promoter occupation by selected proteins was carried out by ChIP-qPCR, while statistical analysis in GraphPad Prism 5 and STATISTICA, ChIP-Seq data were analysed in Galaxy 2.5.0.0. On the list of 28 genes regulated by RBL2, we identified only four solely repressed by RBL2-E2F4-HDAC1-BRM complex. Surprisingly, 24 out of 28 emerged genes controlled by RBL2 were co-regulated by PARP1 in six different manners. In one mode of RBL2/PARP1 co-operation, represented by MAP2K6 and MAPK3, PARP1 was found to associate with gene promoters upon RBL2 silencing, which was previously shown to restore PARP1 expression in monocytes. PARP1 effect on gene transcription was observed only in the presence of active EP300, which acetylated gene promoters and activated transcription. Further analysis revealed that PARP1 binding to MA2K6 and MAPK3 promoters enabled recruitment of EP300 in monocytes, while in proliferating cancer cell lines, which actively transcribe PARP1, this protein maintained EP300 at the promoters of MA2K6 and MAPK3. Genome-wide analysis revealed a similar distribution of PARP1 and EP300 around transcription start sites and the co-occupancy of some gene promoters by PARP1 and EP300 in cancer cells. Here, we described a new RBL2/PARP1/EP300 axis which controls gene transcription regardless of the cell type. In this model cell, cycle-dependent transcription of PARP1 regulates expression of some genes repressed by RBL2 upon cell cycle limitation. Thus, RBL2 may indirectly regulate transcription of some genes by controlling the expression of EP300-recruiting PARP1. Acknowledgement: This work was financed by Polish National Science Centre grants nr DEC-2013/11/D/NZ2/00033 and DEC-2015/19/N/NZ2/01735. L.V. is funded by the National Research, Development and Innovation Office grants GINOP-2.3.2-15-2016-00020 TUMORDNS, GINOP-2.3.2-15-2016-00048-STAYALIVE and OTKA K112336. AR is supported by Polish Ministry of Science and Higher Education 776/STYP/11/2016.

Keywords: retinoblastoma transcriptional co-repressor like 2 (RBL2), poly(ADP-ribose) polymerase 1 (PARP1), E1A binding protein p300 (EP300), monocytes

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Authors: Abdelazeem Algammal, Reham Abouelmaatti, Xiaokun Li, Jisheng Ma, Eman Abdelnaby, Wael Elfeil

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Toll-like receptors (TLRs) are the best understood of the innate immune receptors that detect infections in vertebrates. However, the fish TLRs also exhibit very distinct features and a large diversity, which is likely derived from their diverse evolutionary history and the distinct environments that they occupy. Little is known about the fish immune system structure. Our work was aimed to identify and clone the Nile tilapiaTLR-3 as a model of freshwater fish species; we cloned the full-length cDNA sequence of Nile tilapia (Oreochromis niloticus) TLR-3 and according to our knowledge, it is the first report illustrating tilapia TLR-3. The complete cDNA sequence of Nile tilapia TLR-3 was 2736 pair base and it encodes a polypeptide of 912 amino acids. Analysis of the deduced amino acid sequence indicated that Nile tilapia TLR-3 has typical structural features and main components of proteins belonging to the TLR family. Our results illustrate a complete and functional Nile tilapia TLR-3 and it is considered an ortholog of the other vertebrate’s receptor.

Keywords: Nile tilapia, TLR-3, cloning, gene expression

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Authors: Asma Ben Hmidene, Mizuho Hanaki, Kazuma Murakami, Kazuhiro Irie, Hiroko Isoda, Hideyuki Shigemori

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Type 2 diabetes mellitus (T2DM) and Alzheimer’s disease (AD) are characterized as a peripheral metabolic disorder and a degenerative disease of the central nervous system, respectively. It is now widely recognized that T2DM and AD share many pathophysiological features including glucose metabolism, increased oxidative stress and amyloid aggregation. Amyloid beta (Aβ) is the components of the amyloid deposits in the AD brain and while the component of the amyloidogenic peptide deposit in the pancreatic islets of Langerhans is identified as human islet amyloid polypeptide (hIAPP). These two proteins are originated from the amyloid precursor protein and have a high sequence similarity. Although the amino acid sequences of amyloidogenic proteins are diverse, they all adopt a similar structure in aggregates called cross-beta-spine. Add at that, extensive studies in the past years have found that like Aβ1-42, IAPP forms early intermediate assemblies as spherical oligomers, implicating that these oligomers possess a common folding pattern or conformation. These similarities can be used in the search for effective pharmacotherapy for DM, since potent therapeutic agents such as antioxidants with a catechol moiety, proved to inhibit Aβ aggregation, may play a key role in the inhibit the aggregation of hIAPP treatment of patients with DM. Tamarix gallica is one of the halophyte species having a powerful antioxidant system. Although it was traditionally used for the treatment of various liver metabolic disorders, there is no report about the use of this plant for the treatment or prevention of T2DM and AD. Therefore, the aim of this work is to investigate their protective effect towards T2DM and AD by isolation and identification of α-glucosidase inhibitors, with antioxidant potential, that play an important role in the glucose metabolism in diabetic patient, as well as, the polymerization of hIAPP and Aβ aggregation inhibitors. Structure-activity relationship study was conducted for both assays. And as for α-glucosidase inhibitors, their mechanism of action and their synergistic potential when applied with a very low concentration of acarbose were also suggesting that they can be used not only as α-glucosidase inhibitors but also be combined with established α-glucosidase inhibitors to reduce their adverse effect. The antioxidant potential of the purified substances was evaluated by DPPH and SOD assays. Th-T assay using 42-mer amyloid β-protein (Aβ42) for AD and hIAPP which is a 37-residue peptide secreted by the pancreatic β –cells for T2DM and Transmission electronic microscopy (TEM) were conducted to evaluate the amyloid aggragation of the actives substances. For α-glucosidase, p-NPG and glucose oxidase assays were performed for determining the inhibition potential and structure-activity relationship study. The Enzyme kinetic protocol was used to study the mechanism of action. From this research, it was concluded that polyphenols playing a role in the glucose metabolism and oxidative stress can also inhibit the amyloid aggregation, and that substances with a catechol and glucuronide moieties inhibiting amyloid-β aggregation, might be used to inhibit the aggregation of hIAPP.

Keywords: α-glucosidase inhibitors, amyloid aggregation inhibition, mechanism of action, polyphenols, structure activity relationship, synergistic potential, tamarix gallica

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Authors: Soualem Mami Zoubida, Brixi Nassima, Beghdad Choukri, Belarbi Meriem

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Cereals are the most important source of dietary fiber in the Nordic diet. The fiber in cereals is located mainly in the outer layers of the kernel; particularly in the bran. Improved diet can help unlock the door to good health. Whole grains are an important source of nutrients that are in short supply in our diet, including digestible carbohydrates, dietary fiber, trace minerals, and other compounds of interest in disease prevention, including phytoestrogens and antioxidants (1). The objective of this study is to know the composition of whole grain cereals (rye, millet, white, and red sorghum) which a majority pushes in the south of Algeria. This shows that the millet has a high rate of the sugar estimated at 67.6%. The high proportion of proteins has been found in the two varieties of sorghum and rye. The millet presents the great percentage in lipids compared with the others cereals. And at the last, a red sorghum has the highest rate of fiber(2). These nutrients, as well as other components of whole grain cereals, have, in terms of health, an increased effect if they are consumed together.

Keywords: chemical composition, miller, Secale cereal, Sorghum bicolor

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

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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

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Authors: Azza A. Ali, Doaa M. Abd El-Latif, Amany M. Gad, Yasser M. A. Elnahas, Karema Abu-Elfotuh

Abstract:

Background: Exposure to Aluminium (Al) has been increased recently. It is found in food products, food additives, drinking water, cosmetics and medicines. Chronic consumption of Al causes oxidative stress and has been implicated in several chronic disorders. Liver is considered as the major site for detoxification while kidney is involved in the elimination of toxic substances and is a target organ of metal toxicity. Social isolation (SI) or protein malnutrition (PM) also causes oxidative stress and has negative impact on Al-induced nephrotoxicity as well as hepatotoxicity. Coenzyme Q10 (CoQ10) is a powerful intracellular antioxidant with mitochondrial membrane stabilizing ability while wheat grass is a natural product with antioxidant, anti-inflammatory and different protective activities, cocoa is also potent antioxidants and can protect against many diseases. They provide different degrees of protection from the impact of oxidative stress. Objective: To study the impact of social isolation together with Protein malnutrition on nephro- and hepato-toxicity induced by chronic Al exposure in rats as well as to investigate the postulated protection using a combination of Co Q10, wheat grass and cocoa. Methods: Eight groups of rats were used; four served as protected groups and four as un-protected. Each of them received daily for five weeks AlCl3 (70 mg/kg, IP) for Al-toxicity model groups except one group served as control. Al-toxicity model groups were divided to Al-toxicity alone, SI- associated PM (10% casein diet) and Al- associated SI&PM groups. Protection was induced by oral co-administration of CoQ10 (200mg/kg), wheat grass (100mg/kg) and cocoa powder (24mg/kg) combination together with Al. Biochemical changes in total bilirubin, lipids, cholesterol, triglycerides, glucose, proteins, creatinine and urea as well as alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), lactate deshydrogenase (LDH) were measured in serum of all groups. Specimens of kidney and liver were used for assessment of oxidative parameters (MDA, SOD, TAC, NO), inflammatory mediators (TNF-α, IL-6β, nuclear factor kappa B (NF-κB), Caspase-3) and DNA fragmentation in addition to evaluation of histopathological changes. Results: SI together with PM severely enhanced nephro- and hepato-toxicity induced by chronic Al exposure. Co Q10, wheat grass and cocoa combination showed clear protection against hazards of Al exposure either alone or when associated with SI&PM. Their protection were indicated by the significant decrease in Al-induced elevations in total bilirubin, lipids, cholesterol, triglycerides, glucose, creatinine and urea levels as well as ALT, AST, ALP, LDH. Liver and kidney of the treated groups also showed significant decrease in MDA, NO, TNF-α, IL-6β, NF-κB, caspase-3 and DNA fragmentation, together with significant increase in total proteins, SOD and TAC. Biochemical results were confirmed by the histopathological examinations. Conclusion: SI together with PM represents a risk factor in enhancing nephro- and hepato-toxicity induced by Al in rats. CoQ10, wheat grass and cocoa combination provide clear protection against nephro- and hepatotoxicity as well as the consequent degenerations induced by chronic Al-exposure even when associated with the risk of SI together with PM.

Keywords: aluminum, nephrotoxicity, hepatotoxicity, isolation and protein malnutrition, coenzyme Q10, wheatgrass, cocoa, nutrients combinations

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Authors: Jinyoung Choi, Sunmook Lee

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In general, single parameter, i.e. temperature, as an accelerating parameter is used to assess the accelerated stability of Point-of-Care Testing (POCT) diagnostic devices. However, humidity also plays an important role in deteriorating the strip performance since major components of test strips are proteins such as enzymes. 4 different Temp./Humi. Conditions were used to assess the lifetime of strips. Degradation of test strips were studied through the accelerated stability test and the lifetime was assessed using commercial POCT products. The life distribution of strips, which were obtained by monitoring the failure time of test strip under each stress condition, revealed that the weibull distribution was the most proper distribution describing the life distribution of strips used in the present study. Equal shape parameters were calculated to be 0.9395 and 0.9132 for low and high concentrations, respectively. The lifetime prediction was made by adopting Peck Eq. Model for Stress-Life relationship, and the B10 life was calculated to be 70.09 and 46.65 hrs for low and high concentrations, respectively.

Keywords: accelerated degradation, diagnostic device, lifetime assessment, POCT

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Authors: Sellidj Abdelaziz, Lebaili Soltane

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A cobalt-based alloy type Co-Cr-Ni-WC was deposited by plasma transferred arc projection (PTA) on a stainless steel valve. The alloy is characterized at the equilibrium by a solid solution Co (γ) mainly dendritic, and eutectic carbides M₇C₃ and ηM₆C. At the deposit/substrate interface, this microstructure is modified by the fast cooling mode of the alloy when applied in the liquid state on the relatively cold steel substrate. The structure formed in this case is heterogeneous and metastable phases can occur and evolve over temperature service. Coating properties and reliability are directly related to microstructures formed during deposition. We were interested more particularly in this microstructure formed during the solidification of the deposit in the region of the interface joining the soldered couple and its evolution during cyclic heat treatments at temperatures similar to those of the thermal environment of the valve. The characterization was carried out by SEM-EDS microprobe CAMECA, XRD, and micro hardness profiles. The deposit obtained has a linear and regular appearance that is free of cracks and with little porosity. The morphology of the microstructure represents solidification stages that are relatively fast with a temperature gradient high at the beginning of the interface by forming a plane front solid solution Co (γ). It gradually changes with the decreasing temperature gradient by getting farther from the junction towards the outer limit of the deposit. The matrix takes the forms: cellular, mixed (cells and dendrites) and dendritic. Dendritic growth is done according to primary ramifications in the direction of the heat removal which takes place in the direction perpendicular to the interface, towards the external surface of the deposit, following secondary and tertiary undeveloped arms. The eutectic carbides M₇C₃ and ηM₆C formed are very thin and are located in the intercellular and interdendritic spaces of the solid solution Co (γ).

Keywords: Co-Ni-Cr-W-C alloy, solid deposit, microstructure, carbides, cyclic heat treatment

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Authors: Sidra Islam, Moin Uddin, Mir A. Rouf

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A wide variety of pathological disorders owing to hyperglycemic conditions involves structural rearrangements and condensations of proteins. The implication of methylglyoxal (MG) modified immunoglobulin G (IgG) in the onset and progression of diabetes type 2 (T2DM) is studied in the present study. Using biophysical and biochemical approaches MG was found to perturb the structure of IgG, effect its microenvironment and leads to aggregate formation. Furthermore, MG-IgG was found to be highly immunogenic inducing high titre antibodies in female rabbits. Clinical studies revealed the presence of circulating anti-MG-IgG antibodies as analyzed by direct binding ELISA. The circulating auto antibodies were highly specific for MG-IgG as revealed by inhibition ELISA. Thus it can be concluded that MG is a powerful agent with a high damaging potential. To IgG. It is highly capable of generating immune response that contributes to the immunopathology associated with diabetes. Dicarbonyl adducts may emerge as potential biomarkers for T2DM.

Keywords: immunogenicity, Immunoglobulin G, methylglyoxal, Type 2 Diabetes Mellitus

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Authors: Nichole Haag, Kimberly Velk, Tyler McCune, Chun Wu

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Methicillin/multiple-resistant Staphylococcus aureus (MRSA) are infectious bacteria that are resistant to common antibiotics. A previous in silico study in our group has identified a hypothetical protein SAV1226 as one of the potential drug targets. In this study, we reported the bioinformatics characterization, as well as cloning, expression, purification and kinetic assays of hypothetical protein SAV1226 from methicillin/vancomycin-resistant Staphylococcus aureus Mu50 strain. MALDI-TOF/MS analysis revealed a low degree of structural similarity with known proteins. Kinetic assays demonstrated that hypothetical protein SAV1226 is neither a domain of an ATP dependent dihydroxyacetone kinase nor of a phosphotransferase system (PTS) dihydroxyacetone kinase, suggesting that the function of hypothetical protein SAV1226 might be misannotated on public databases such as UniProt and InterProScan 5.

Keywords: Methicillin-resistant Staphylococcus aureus, dihydroxyacetone kinase, essential genes, drug target, phosphoryl group donor

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Authors: Sema Şenoğlu, Sevgi Karakuş

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Hydrazone scaffold is important to design new drug groups and is found to possess numerous uses in pharmaceutical chemistry. Besides, hydrazone derivatives are also known for biological activities such as anticancer, antimicrobial, antiviral, and antifungal. Hydrazone derivatives are promising anticancer agents because they inhibit cancer proliferation and induce apoptosis. Human carbonic anhydrase IX has a high potential to be an antiproliferative drug target, and targeting this protein is also important for obtaining potential anticancer inhibitors. The protein construct was retrieved as a PDB file from the RCSB protein database. This binding interaction of proteins and ligands was performed using Discovery Studio Visualizer. In vitro inhibitory activity of hydrazone derivatives was tested against enzyme carbonic anhydrase IX on the PyRx programme. Most of these molecules showed remarkable human carbonic anhydrase IX inhibitory activity compared to the acetazolamide. As a result, these compounds appear to be a potential target in drug design against human carbonic anhydrase IX.

Keywords: cancer, carbonic anhydrase IX enzyme, docking, hydrazone

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Authors: Ihtisham Bukhari

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In the current study, we enrolled a 46, XY phenotypically female patient bearing testes in her inguinal canal. DNA sequencing of the AR gene detected a missense mutation C.1715A > G (p. Y572C) in exon 2 which is already known to cause Complete androgen insensitivity syndrome (CAIS). We further studied the effects of this mutation on the testicular histopathology of the patient. No spermatocytes were seen in the surface spreading of testicular tissues while H&E staining showed that seminiferous tubules predominantly have only Sertoli cells. To confirm this meiotic failure is likely due to the current AR mutation we performed mRNA expression of genes associated with AR pathway, expression and location of the associated proteins in testicular tissues. Western blot and real-time PCR data showed that the patient had high levels of expression of AMH, SOX9, and INNB in testis. Tubules were stained with SOX9 and AMH which revealed Sertoli cell maturation arrest. Therefore, we suggest that AR mutation enhances AMH expression which ultimately leads to failure in the maturation of Sertoli cells and failure in spermatogenesis.

Keywords: androgen receptor, spermatogenesis, infertility, Sertoli cell only syndrome

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Authors: G. Medkour Ishak-Boushaki, A. Taibi, M. Allab

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Quantitative environmental radioactivity measurements are needed to determine the level of exposure of a population to ionizing radiations and for the assessment of the associated risks. Gamma spectrometry remains a very powerful tool for the analysis of radionuclides present in an environmental sample but the basic problem in such measurements is the low rate of detected events. Using large environmental samples could help to get around this difficulty but, unfortunately, new issues are raised by gamma rays attenuation and self-absorption. Recently, a new method has been suggested, to detect and identify without quantification, in a short time, a gamma ray of a low count source. This method does not require, as usually adopted in gamma spectrometry measurements, a pulse height spectrum acquisition. It is based on a chronological record of each detected photon by simultaneous measurements of its energy ε and its arrival time τ on the detector, the pair parameters [ε,τ] defining an event mode sequence (EMS). The EMS serials are analyzed sequentially by a Bayesian approach to detect the presence of a given radioactive source. The main object of the present work is to test the applicability of this sequential approach in radioactive environmental materials detection. Moreover, for an appropriate health oversight of the public and of the concerned workers, the analysis has been extended to get a reliable quantification of the radionuclides present in environmental samples. For illustration, we consider as an example, the problem of detection and quantification of 238U. Monte Carlo simulated experience is carried out consisting in the detection, by a Ge(Hp) semiconductor junction, of gamma rays of 63 keV emitted by 234Th (progeny of 238U). The generated EMS serials are analyzed by a Bayesian inference. The application of the sequential Bayesian approach, in environmental radioactivity analysis, offers the possibility of reducing the measurements time without requiring large environmental samples and consequently avoids the attached inconvenient. The work is still in progress.

Keywords: Bayesian approach, event mode sequence, gamma spectrometry, Monte Carlo method

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Authors: Venoos Iman, Suzita Mohd Noor, Syam Mohan, Mohamad Ibrahim Noordin

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Girinimbine, a carbazole alkaloid was isolated from the stem bark and root of Murraya koenigii and its structure and purity was identified by HPLC and LC-MS. Here we report that Girinimbine strongly inhibit angiogenesis activity both in vitro and in vivo. MTT result showed that girinimbine inhibits cell proliferation of the HUVECS cell line in vitro. Result of endothelial cell invasion, migration, tube formation and wound healing assays also demonstrated significant time and does dependent inhibition by girinimbine. Moreover, girinibine mediates its anti-angiogenic activity through up- and down-regulation of angiogenic and anti-aniogenic proteins. Furthermore, anti-angiogenic potential of girinimbine was evidenced in vivo on zebrafish model. Girinimbine inhibited neo-vessels formation in zebrafish embryos during 24 hours exposure time. Together, these results demonstrated for the first time that girinimbine could effectively suppress angiogenesis and strongly suggest that it might be a novel angiogenesis inhibitor.

Keywords: anti-angiogenic, carbazole alkaloid, girinimbine, zebrafish

Procedia PDF Downloads 376

Authors: Md. Maksudur Rahman Khan, M. Rahim Uddin, Hamidah Abdullah, Kaykobad Md. Rezaul Karim, Abu Yousuf, Chin Kui Cheng, Huei Ruey Ong

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A systematic study was conducted to explore the photocatalytic reduction of carbon dioxide (CO₂) into methanol on TiO₂ loaded copper ferrite (CuFe₂O₄) photocatalyst under visible light irradiation. The phases and crystallite size of the photocatalysts were characterized by X-ray diffraction (XRD) and it indicates CuFe₂O₄ as tetragonal phase incorporation with anatase TiO₂ in CuFe₂O₄/TiO₂ hetero-structure. The XRD results confirmed the formation of spinel type tetragonal CuFe₂O₄ phases along with predominantly anatase phase of TiO₂ in the CuFe₂O₄/TiO₂ hetero-structure. UV-Vis absorption spectrum suggested the formation of the hetero-junction with relatively lower band gap than that of TiO₂. Photoluminescence (PL) technique was used to study the electron–hole (e⁻/h⁺) recombination process. PL spectra analysis confirmed the slow-down of the recombination of electron–hole (e⁻/h⁺) pairs in the CuFe₂O₄/TiO₂ hetero-structure. The photocatalytic performance of CuFe₂O₄/TiO₂ was evaluated based on the methanol yield with varying amount of TiO₂ over CuFe₂O₄ (0.5:1, 1:1, and 2:1) and changing light intensity. The mechanism of the photocatalysis was proposed based on the fact that the predominant species of CO₂ in aqueous phase were dissolved CO₂ and HCO₃^- at pH ~5.9. It was evident that the CuFe₂O₄ could harvest the electrons under visible light irradiation, which could further be injected to the conduction band of TiO₂ to increase the life time of the electron and facilitating the reactions of CO₂ to methanol. The developed catalyst showed good recycle ability up to four cycles where the loss of activity was ~25%. Methanol was observed as the main product over CuFe₂O₄, but loading with TiO₂ remarkably increased the methanol yield. Methanol yield over CuFe₂O₄/TiO₂ was found to be about three times higher (651 μmol/g_cat L) than that of CuFe₂O₄ photocatalyst. This occurs because the energy of the band excited electrons lies above the redox potentials of the reaction products CO₂/CH₃OH.

Keywords: photocatalysis, CuFe2O4/TiO2, band-gap energy, methanol

Procedia PDF Downloads 244

Authors: Z. Herceg, V. Stulic, T. Vukusic, A. Rezek Jambrak

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High voltage electrical discharge plasmas are new nonthermal developing techniques used for water decontamination. To the full understanding of cell inactivation mechanisms, this study brings inactivation, recovery and cellular leakage of L. innocua cells before and after the treatment. Bacterial solution (200 mL) of L. innocua was treated in a glass reactor with a point-to-plate electrode configuration (high voltage electrode-titanium wire, was in the gas phase and grounded electrode was in the liquid phase). Argon was injected into the headspace of the reactor at the gas flow of 5 L/min. Frequency of 60, 90 and 120 Hz, time of 5 and 10 min, positive polarity and conductivity of media of 100 µS/cm were chosen to define listed parameters. With a longer treatment time inactivation was higher as well as the increase in cellular leakage. Despite total inactivation recovery of cells occurred probably because of a high leakage of proteins, compared to lower leakage of nucleic acids (DNA and RNA). In order to define mechanisms of inactivation further research is needed.

Keywords: Listeria innocua ATCC 33092, inactivation, gas phase plasma, cellular leakage, recovery of cells

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Authors: Ali Ammari, Karin Schroen

Abstract:

In brewing industries, stripping is a downstream process to separate volatiles from beer. Due to physiochemical similarities between flavour components, the selectivity of this method is not favourable. Besides, the presence of non-volatile compounds such as proteins and carbohydrates may affect the separation of flavours due to their retaining properties. By using a stripping column with structured packing coupled with a gas chromatography, in this work, the overall mass transfer coefficient along with their corresponding equilibrium data was investigated for a model solution consist of water, ethanol, ethyl acetate and isoamyl acetate. Static headspace analysis also was employed to derive equilibrium data for flavours in the presence of beer dry matter. As it was expected ethanol and dry matter showed retention properties; however, the effect of viscosity in mass transfer coefficient was discarded due to the fact that the viscosity of solution decreased during stripping. The effect of ethanol and beer dry matter were mapped to be used for designing stripping could.

Keywords: flavour, headspace, Henry’s coefficient, mass transfer coefficient, stripping

Procedia PDF Downloads 194