Search results for: fusion proteins

Authors: Jasna M. Čanadanović-Brunet, Gordana S. Ćetković, Jelena J. Vulić, Sonja M. Djilas, Vesna T. Tumbas Šaponjac, Sladjana M. Stajčić

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Honey serves as a source of natural antioxidants, which are effective in reducing the risk of heart disease, cancer, immune-system decline, cataracts, different inflammatory processes, and also prevent deteriorative oxidation reactions in foods such as enzymatic browning of fruit and vegetables. Honey is a natural saturated sugar solution, but it also contains certain minor constituents, proteins, enzymes, amino and organic acids, lipids, vitamins, phenolic acids, flavonoids and carotenoids. It is consumed in its natural form alone, but also in combination with nuts and various kinds of dried fruits. The aim of this research was to investigate the contribution of dried cherry on phenols (TPh) and flavonoids (Fl) contents and antioxidant activities of honey. Phenolic compounds in Serbian polyfloral (PH), linden (LH) and acacia (AH) honey and also in their mixtures with dried cherry, in 40% mass concentrations (PH40; LH40, AH40), were determined. In comparison to honey, TPh increased 2.25 times for LH40, 2.16 times for AH40 and 1.45 times for PH40, while Fl increased 2.81-fold for PH40, 1.21-fold for LH40 and 1.44-fold for AH40. Antioxidant activity was investigated with two assays, DPPH test and reducing power (RP), and expressed as EC50DPPH and RP0.5 values. The EC50DPPH values were: EC50PH40 = 1.16 mg/ml; EC50LH40= 1.42 mg/ml and EC50AH40= 1.69 mg/ml, while RP0.5 were: RP0.5PH40 = 15.05 mg/ml; RP0.5LH40 = 16.09 mg/ml and P0.5AH40 = 17.60 mg/ml. Our results indicate that supplementation of polyfloral, linden and acacia honey with 40% dried cherry improves antioxidant activity of honey by enriching the phenolic composition.

Keywords: antioxidant activity, dried cherry, honey, phenolics

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Authors: Agbugui M. O., Inobeme A.

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Fish constitutes a vital component of human diets due to their rich nutritional compositions. They serve as a remarkable source of proteins, vitamins, and fatty acids, which are indispensable for the effective growth and development of humans. The need to explore the nutritional compositions of various species of fish in different water bodies becomes paramount. Presently, consumer concern is not just on food's nutritional value but also on the safety level. Environmental contamination by heavy metals has become an issue of pressing concern in recent times. Heavy metals, due to their ubiquitous nature, are found in various water bodies as they are released from various anthropogenic activities. This work investigated the proximate compositions, mineral contents, and heavy metals concentrations of four different species of fish (P. annectens, L. niloticus, G. niloticus, and H. niloticus) collected from the lower Niger at Agenebode using standard procedures. The highest protein contents were in Gymnarchus niloticus (37.32%), while the least was in Heterotis niloticus (20.41%). Protopterus annectens had the highest carbohydrate content (34.55%), while Heterotis niloticus had the least (12.24%). The highest lipid content (14.41%) was in Gymnarchus niloticus. The highest concentration of potassium was 21.00 ppm. The concentrations of heavy metals in ppm ranged from 0.01 – 1.4 (Cd), 0.07 – 2.89 (Pb), 0.02 – 16.4 (Hg), 0.88 – 5.1 (Cu) and 1.2 – 8.23 (Zn). The concentrations of Hg, Cd and Pb in some of the samples investigated were higher than the permissible limits based on international standards. There is a pressing need for further study focusing on various species of animals and plants in the area due to the alarming contents of these metals; remedial measures could also be ensured for safety.

Keywords: trace metals, nutritional value, human health, crude protein, lipid content

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Authors: Mateen A. Khan, Elizabeth J. Theil, Dixie J. Goss

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Cellular iron homeostasis is accomplished by the coordinated regulated expression of iron uptake, storage, and export. Iron regulate the translation of ferritin and mitochondrial aconitase iron responsive element (IRE)-mRNA by interaction with an iron regulatory protein (IRPs). Iron increases protein biosynthesis encoded in iron responsive element. The noncoding structure IRE-mRNA, approximately 30-nt, folds into a stem loop to control synthesis of proteins in iron trafficking, cell cycling, and nervous system function. Fluorescence anisotropy measurements showed the presence of one binding site on IRP1 for ferritin and mitochondrial aconitase IRE-mRNA. Scatchard analysis revealed the binding affinity (Kₐ) and average binding sites (n) for ferritin and mitochondrial aconitase IRE-mRNA were 68.7 x 10⁶ M⁻¹ and 9.2 x 10⁶ M⁻¹, respectively. In order to understand the relative importance of equilibrium and stability, we further report the contribution of electrostatic interactions in the overall binding of two IRE-mRNA with IRP1. The fluorescence quenching of IRP1 protein was measured at different ionic strengths. The binding affinity of IRE-mRNA to IRP1 decreases with increasing ionic strength, but the number of binding sites was independent of ionic strength. Such results indicate a differential contribution of electrostatics to the interaction of IRE-mRNA with IRP1, possibly related to helix bending or stem interactions and an overall conformational change. Selective destabilization of ferritin and mitochondrial aconitase RNA/protein complexes as reported here explain in part the quantitative differences in signal response to iron in vivo and indicate possible new regulatory interactions.

Keywords: IRE-mRNA, IRP1, binding, ionic strength

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Authors: Seyyed Mohammad Amin Mousavi Sagharchi, Elham Javanroudi

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Introduction: Tuberculosis (TB) is a known disease with hidden features caused by Mycobacterium tuberculosis (MTB). This disease, which is one of the 10 deadliest in the world, has caused millions of deaths in recent decades. Furthermore, TB is responsible for infecting about 30% population of world. Like any infection, TB can activate the immune system by locating and colonization in the human body, especially in the alveoli. TB is granulomatosis, so MTB can absorb the host’s immune cells and other cells to form granuloma. Method: Different databases (e.g., PubMed) were recruited to prepare this paper and fulfill our goals to search and find effective papers and investigations. Results: Immune response to MTB is related to T cell killers and contains CD1, CD4, and CD8 T lymphocytes. CD1 lymphocytes can recognize glycolipids, which highly exist in the Mycobacterial fatty cell wall. CD4 lymphocytes and macrophages form granuloma, and it is the main line of immune response to Mycobacteria. On the other hand, CD8 cells have cytolytic function for directly killing MTB by secretion of granulysin. Other functions and secretion to the deal are interleukin-12 (IL-12) by induction of expression interferon-γ (INF-γ) for macrophages activation and creating a granuloma, and tumor necrosis factor (TNF) by promoting macrophage phagolysosomal fusion. Conclusion: Immune cells in battle with MTB are macrophages, dendritic cells (DCs), neutrophils, and natural killer (NK) cells. These immune cells can recognize the Mycobacterium by various receptors, including Toll-like receptors (TLRs), Nod-like receptors (NLRs), and C-type lectin receptors (CLRs) located in the cell surface. In human alveoli exist about 50 dendritic macrophages, which have close communication with other immune cells in the circulating system and epithelial cells to deal with Mycobacteria. Against immune cells, MTB handles some factors (e.g., cordfactor, O-Ag, lipoarabinomannan, sulfatides, and adenylate cyclase) and practical functions (e.g., inhibition of macrophages).

Keywords: mycobacterium tuberculosis, immune responses, immunological mechanisms, respiratory tuberculosis

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Authors: Alireza Naseri, Susan L. Holdt, Charlotte Jacobsen

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In 2014, the global amount of carrageenan production was 60,000 ton with a value of US$ 626 million. From this number, it can be estimated that the total dried seaweed consumption for this production was at least 300,000 ton/year. The protein content of these types of seaweed is 5 – 25%. If just half of this total amount of protein could be extracted, 18,000 ton/year of a high-value protein product would be obtained. The overall aim of this study was to develop a technology that will ensure further utilization of the seaweed that is used only as raw materials for carrageenan production as single extraction at present. More specifically, proteins should be extracted from the seaweed either before or after extraction of carrageenan with focus on maintaining the quality of carrageenan as a main product. Different mechanical, chemical and enzymatic technologies were evaluated. The optimized process was implemented in lab scale and based on its results; the new experiments were done a pilot and larger scale. In order to calculate the efficiency of the new upstream multi-extraction process, protein content was tested before and after extraction. After this step, the extraction of carrageenan was done and carrageenan content and the effect of extraction on yield were evaluated. The functionality and quality of carrageenan were measured based on rheological parameters. The results showed that by using the new multi-extraction process (submitted patent); it is possible to extract almost 50% of total protein without any negative impact on the carrageenan quality. Moreover, compared to the routine carrageenan extraction process, the new multi-extraction process could increase the yield of carrageenan and the rheological properties such as gel strength in the final carrageenan had a promising improvement. The extracted protein has initially been screened as a plant protein source in typical food applications. Further work will be carried out in order to improve properties such as color, solubility, and taste.

Keywords: carrageenan, extraction, protein, seaweed

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Authors: Jaruwan Duangchuen, Siwalak Pathaveerat

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Coconut (Cocos nucifera) belongs to the family Arecaceae. Coconut juice and meat are consumed as food and dessert in several regions of the world. Coconut juice contains low proteins, and arginine is the main amino acid content. Coconut meat is the endosperm of coconut that has nutritional value. It composes of carbohydrate, protein and fat. The objective of this study is utilization of by-products from the virgin coconut oil extraction process by using the skimmed coconut milk as a powder. The skimmed coconut milk was separated from the coconut milk in virgin coconut oil extraction process that consists approximately of protein 6.4%, carbohydrate 7.2%, dietary fiber 0.27 %, sugar 6.27%, fat 3.6 % and moisture content of 86.93%. This skimmed coconut milk can be made to powder for value - added product by using spray drying. The factors effect to the yield and properties of dry skimmed coconut milk in spraying process are inlet, outlet air temperature and the maltodextrin concentration. The percentage of maltodextrin content (15, 20%), outlet air temperature (80 ºC, 85 ºC, 90 ºC) and inlet air temperature (190 ºC, 200 ºC, 210 ºC) were conducted to the skimmed coconut milk spray drying process. The spray dryer was kept air flow rate (0.2698 m3 /s). The result that shown 2.22 -3.23% of moisture content, solubility, bulk density (0.4-0.67g/mL), solubility, wettability (4.04 -19.25 min) for solubility in the water, color, particle size were analyzed for the powder samples. The maximum yield (18.00%) of spray dried coconut milk powder was obtained at 210 °C of temperature, 80°C of outlet temperature and 20% maltodextrin for 27.27 second for drying time. For the amino analysis shown that the high amino acids are Glutamine (16.28%), Arginine (10.32%) and Glycerin (9.59%) by using HPLP method (UV detector).

Keywords: skimmed coconut milk, spray drying, virgin coconut oil process (VCO), maltodextrin

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Authors: Zakaria Boual, Abdellah Kemassi, Toufik Chouana, Philippe Michaud, Mohammed Didi Ould El Hadj

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In order to investigate the prebiotic potential of oligosaccharides prepared by chemical hydrolysis of water-soluble polysaccharides (WSP) from Zizyphus lotus leaves, the effect of oligosaccharides on bacterial growth was studied. The chemical composition of WSP was evaluated by colorimetric assays revealed the average values: 7.05±0.73% proteins and 86.21±0.74% carbohydrates, among them 64.81±0.42% are neutral sugar and the rest 16.25±1.62% are uronic acids. The characterization of monosaccharides was determined by high performance anion exchange chromatography with pulsed amperometric detection (HPAEC-PAD) was found to be composed of galactose (23.95%), glucose (21.30%), rhamnose (20.28%), arabinose (9.55%), and glucuronic acid (22.95%). The effects of oligosaccharides on the growth of lactic acid bacteria were compared with those of fructo-oligosaccharide (RP95). The oligosaccharides concentration was 1g/L of man rogosa sharpe broth. Bacterial growth was assessed during 2, 4.5, 6.5, 9, 12, 16 and 24 h by measuring the optical density of the cultures at 600 nm (OD600) and pH values. During fermentation, pH in broth cultures decreased from 6.7 to 5.87±0.15. The enumeration of lactic acid bacteria indicated that oligosaccharides led to a significant increase in bacteria (P≤0.05) compared to the control. The fermentative metabolism appeared to be faster on RP95 than on oligosaccharides from Zizyphus lotus leaves. Both RP95 and oligosaccharides showed clear prebiotic effects, but had differences in fermentation kinetics because of to the different degree of polymerization. This study shows the prebiotic effectiveness of oligosaccharides, and provides proof for the selection of leaves of Zizyphus lotus for use as functional food ingredients.

Keywords: Zizyphus lotus, polysaccharides, characterization, prebiotic effects

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Authors: Shima Mahmoudi, Babak Pourakbari, Setareh Mamishi, Mostafa Teymuri, Majid Marjani

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Although cytokine analysis has greatly contributed to the understanding of tuberculosis (TB) pathogenesis, data on cytokine profiles that might distinguish progression from latency of TB infection are scarce. Since PE/PPE proteins are known to induce strong humoral and cellular immune responses, the aim of this study was to evaluate the diagnostic potential of interleukin-2 (IL-2) as biomarker after specific antigen stimulation with PE35 and PPE68 for the discrimination of active and latent tuberculosis infection (LTBI). The production of IL-2 was measured in the antigen-stimulated whole-blood supernatants following stimulation with recombinant PE35 and PPE68. All the patients with active TB and LTBI had positive QuantiFERON-TB Gold in Tube test. The level of IL-2 following stimulation with recombinant PE35 and PPE68 were significantly higher in LTBI group than in patients with active TB infection or control group. The discrimination performance (assessed by the area under ROC curve) for IL-2 following stimulation with recombinant PE35 and PPE68 between LTBI and patients with active TB were 0.837 (95%CI: 0.72-0.97) and 0.75 (95%CI: 0.63-0.89), respectively. Applying the 12.4 pg/mL cut-off for IL-2 induced by PE35 in the present study population resulted in sensitivity of 78%, specificity of 78%, PPV of 78% and NPV of 100%. In addition, a sensitivity of 81%, specificity of 70%, PPV of 67% and 87% of NPV was reported based on the 4.4 pg/mL cut-off for IL-2 induced by PPE68. In conclusion, peptides of the antigen PE35 and PPE68, absent from commonly used BCG strains, stimulated strong IL-2- positive T cell responses in patients with LTBI. This study confirms IL-2 induced by PE35 and PPE68 as a sensitive and specific biomarker and highlights IL-2 as new promising adjunct markers for discriminating of LTBI and Active TB infection.

Keywords: IL-2, PE35, PPE68, tuberculosis

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Authors: Irma Gabriela Lopez-Moreno, Elva Perez-Luque, Herlinda Aguilar-Zavala

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Introduction: Type 2 Diabetes Mellitus (T2DM) is characterized by combination of insulin resistance and deterioration of insulin secretion. Sfrp5 is a protein that antagonizes Wnt5a proteins by preventing it from reaching its receptor and activating the Wnt/β-catenin signaling pathway, this pathway is one of the most important regulators of adipogenesis. Although metformin decreases glucose levels its mechanisms of action are not fully known but it has been implicated in the inhibition of the Wnt/β-catenin signaling pathway. Objective: The objective was evaluating the effects of metformin on serum levels of Sfrp5 in patients with T2DM treated and not treated with metformin. Methods: Two groups of patients were selected: one group of T2DM patients treated with metformin (n = 35) and another group of subjects with recent diagnosis of T2DM untreated (n = 35) with a mean age of 48 ± 9 years. In these subjects anthropometric measures were taken as weight, height, waist and hip circumference, were calculated the percentage of body fat, visceral fat and muscle mass. In addition, were measured glucose levels, lipid profile, adiponectin and Sfrp5. Results: Sfrp5 were higher in metformin-treated patients compared to the untreated group (19.9 vs 13.6 ng/mL p < 0.001), a negative correlation was found between Sfrp5 levels and total cholesterol levels (r= -0.25, p = 0.03) and percentage of visceral fat (r = -0.26, p = 0.03) and a positive correlation with HDL cholesterol levels (r = 0.31, p = 0.01) and adiponectin (r=0.65, p = < 0.001). Conclusions: The findings show that metformin consumption increased levels of Sfrp5, which may lead to a decrease in the activation of the WNT/β-catenin pathway impacting on adipogenesis.

Keywords: adiponectin, diabetes, metformin, Sfrp5

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Authors: Hawbash Jalil

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In this study, the effects of whey protein based films on various properties of kashar cheese were examined. In the study, edible film solutions based on whey protein isolate, whey protein isolate + transglutaminase enzyme and whey protein isolate + chitosan were produced and Kashar cheese samples were coated with these films by dipping method and stored at +4 ºC for 60 days. Chemical, microbiological and textural analyzes were carried out on samples at 0, 30 and 60 days of storage. As a result of the study, the highest dry matter and total nitrogen values were obtained from uncoated control samples This is an indication that the coatings limit water vapor permeability. The highest acidity and pH values obtained from the samples as storage results were 3.33% and 5.86%, respectively, in the control group samples. Both acidity and pH rise in these groups, is a consequence of the buffering of pH changes of hydrolsis products which are as a result of proteolysis occurring in the sample. Nitrogen changes and lipolysis values, which are indicative of the degree of hydrolysis of proteins and triglycerides in kashar cheese, were generally higher in the control group This result is due to limiting the micro organism reproduction by limiting the gas passage of the coatings. Hardness and chewiness values of the textural properties of the samples were significantly reduced in uncoated control samples compared to the coated samples due to maturation. The chitosan film coatings used in the study limited the development of mold yeast until the 30th day but after that did not yield successful results in this respect.

Keywords: chitosan, edible film, transglutaminase, whey protein

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Authors: M. Cherki, I. Taouam, A. Amiri, F. Hmimid, T. Ould Bellahcen

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The Moroccan coasts represent an important wealth of red algae which have an economic interest. Among these algae, the Gelidium sesquipedale, which is exploited industrially for its richness in agar. The aim of this study is to establish a general overview of the macronutrient composition of Gelidium sesquipedale and to compare this composition according to three factors: the harvest site (El Jadida, Casablanca and Mohammadia), the harvest depth (coast and depth) and the drying process (open air and oven). Proteins, lipids, and carbohydrates are measured by different methods. The analysis of results show that the protein concentrations of the El Jadida and Mohammadia samples are significantly higher than that of Casablanca (0.026 ± 0.0007 µg/µg DW 0.024 ± 0.001 µg/µg DW and 0.006 ± 0.0007 µg/µg DW, p < 0.05 respectively). However, Casablanca samples are significantly richer in total sugars (0.023 ± 0.002 µg/µg DW, p < 0.05) and less rich in reducing sugars (0.0001 ± 0.00001 µg/µg DW, p < 0.05) compared to other samples. The lipid concentrations of the samples from the three harvest sites do not show any significant difference. With respect to depth, only total protein and total sugar concentrations were significantly higher in the coast versus depth samples (0.035 ± 0.004 µg/µg DW vs. 0.026 ± 0.0007 µg/µg DW and 0.035 ± 0.006 µg/µg DW vs. 0.012 ± 0.005 µg/µg DW p < 0.05 respectively). For the drying process, protein, total sugars and lipid concentrations were significantly higher in open air samples compared to oven samples (0.006 ± 0.0007 µg/µg DW). vs 0.004 ± 0.0003 µg/µg DW, 0.023 ± 0.002 µg/µg DW vs 0.007 ± 0.002 µg/µg DW and 8% vs 4% p < 0.05 respectively). Our results demonstrate that the chemical composition of Gelidium sesquipedale varies according to the harvest region. In addition, samples harvested on the coast and dried in the open air are the richest in macronutrients.

Keywords: biochemical composition, drying, depth, Gelidium sesquipedale, red algae, region

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Authors: Dipranjan Laha, Parimal Karmakar

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Metal oxide nanoparticles are well known to generate oxidative stress and deregulate normal cellular activities. Among these, transition metals copper oxide nanoparticles (CuO NPs) are more compelling than others and able to modulate different cellular responses. In this work, we have synthesized and characterized CuO NPs by various biophysical methods. These CuO NPs (~30 nm) induce autophagy in human breast cancer cell line, MCF7 in a time and dose-dependent manner. Cellular autophagy was tested by MDC staining, induction of green fluorescent protein light chain 3 (GFP-LC3B) foci by confocal microscopy, transfection of pBABE-puro mCherry-EGFP-LC3B plasmid and western blotting of autophagy marker proteins LC3B, beclin1, and ATG5. Further, inhibition of autophagy by 3-Methyladenine (3-MA) decreased LD50 doses of CuO NPs. Such cell death was associated with the induction of apoptosis as revealed by FACS analysis, cleavage of PARP, dephosphorylation of Bad and increased cleavage product of caspase3. siRNA-mediated inhibition of autophagy-related gene beclin1 also demonstrated similar results. Finally, induction of apoptosis by 3-MA in CuO NPs treated cells were observed by TEM. This study indicates that CuO NPs are a potent inducer of autophagy which may be a cellular defense against the CuO NPs mediated toxicity and inhibition of autophagy switches the cellular response into apoptosis. A combination of CuO NPs with the autophagy inhibitor is essential to induce apoptosis in breast cancer cells. Acknowledgments: The authors would like to acknowledge for financial support for this research work to the Department of Biotechnology (No. BT/PR14661/NNT/28/494/2010), Government of India.

Keywords: nanoparticle, autophagy, apoptosis, siRNA-mediated inhibition

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Authors: Abdullah A. Al Qurashi, Hattan A. Hassani, Bader K. Alaslap

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Arrhythmogenic Right Ventricular Dysplasia/Cardiomyopathy (ARVD/C) is an uncommon, inheritable cardiac disorder characterized by the progressive substitution of cardiac myocytes by fibro-fatty tissues. This pathologic substitution predisposes patients to ventricular arrhythmias and right ventricular failure. The underlying genetic defect predominantly involves genes encoding for desmosome proteins, particularly plakophilin-2 (PKP2). These aberrations lead to impaired cell adhesion, heightening the susceptibility to fibrofatty scarring under conditions of mechanical stress. Primarily, ARVD/C affects the right ventricle, but it can also compromise the left ventricle, potentially leading to biventricular heart failure. Clinical presentations can vary, spanning from asymptomatic individuals to those experiencing palpitations, syncopal episodes, and, in severe instances, sudden cardiac death. The establishment of a diagnostic criterion specifically tailored for ARVD/C significantly aids in its accurate diagnosis. Nevertheless, the task of early diagnosis is complicated by the disease's frequently asymptomatic initial stages, and the overall rarity of ARVD/C cases reported globally. In some cases, as exemplified by the adult female patient in this report, the disease may advance to terminal stages, rendering therapies like Ventricular Tachycardia (VT) ablation ineffective. This case underlines the necessity for increased awareness and understanding of ARVD/C to aid in its early detection and management. Through such efforts, we aim to decrease morbidity and mortality associated with this challenging cardiac disorder.

Keywords: arrhythmogenic right ventricular dysplasia, cardiac disease, interventional cardiology, cardiac electrophysiology

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Authors: Mohammed Althobiti, Patrick Booms, Dorota Fiszer, Philip Hopkins

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Malignant hyperthermia (MH) is a pharmacogenetic disorder of skeletal muscle. MH results from anaesthetics induced breakdown of calcium homeostasis. RYR1 and CACN1AS mutations represent the aetiology in ~70% of the MH population. Previous studies indicate that up to 25% of MH patients carry no variants in these genes. Therefore, the aim of this study is to investigate the relationships between MH susceptibility and genes encoding skeletal muscle Ca2+ channels as well as accessory proteins. The JSRP, encoding JP-45, was previously sequenced and novel genetic variants were identified. The variant p.P108L (c.323C > T) was identified in exon 4 and encodes a change from a proline at amino acid 108 to leucine residue. The variant P108L was detected in two patients out of 50 with 4% frequency in the sample population. The alignment of DNA sequences in different species indicates highly conserved proline sequences involved in the substitution of the P108L variant. In this study, the variant P108L co-segregates with the SNP p.V92A (c.275T > C) at the same exon, both variants being inherited in the same two patients only. This indicates that the two variants may represent a haplotype. Therefore, a set of single nucleotide polymorphisms and statistical analysis will be used to investigate the effects of haplotypes on MH susceptibility. Furthermore, investigating the effect of the P108L variant in combination with RYR1 mutations or other genetic variants in other genes as a combination of two or more genetic variants, haplotypes may then provide stronger genetic evidence indicating that JSRP1 is associated with MH susceptibility. In conclusion, these preliminary results lend a potential modifier role of the variant P108L in JSRP1 in MH susceptibility and further investigations are suggested to confirm these results.

Keywords: JSRP1, malignant hyperthermia, RyR1, skeletal muscle

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Authors: Youjeong Hwang

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Purpose: Insulin-like growth factor-I (IGF-I) promotes B-cell development, immunoglobulin formation, and interleukin-6 (IL-6) production, then regulate the immune response and inflammation. As IGF-I and their receptor also exist in the periodontal tissue, they may affect the immune response caused by periodontal pathogens in aggressive periodontitis (AgP) patients. The function of IGF is regulated by IGF binding proteins (IGFBPs), and IGFBP-3 is known to most abundant in plasma. The aim of the present study was to assess the concentration of IGF-I and IGFBP-3 in plasma and gingival crevicular fluid (GCF) in AgP patients and to find out their association. Methods: Nine patients with AgP (test group) and nine healthy subjects (control group) were included in this study. None of the subjects had a history of systemic disease, smoking or steroids medication. GCF samples were collected by microcapillary pipettes and plasma samples were obtained by venipuncture. Probing pocket depth (PD), clinical attachment level (CAL) and bleeding on probing (BOP) were recorded. Samples were assayed for IGF-I and IGFBP-3 levels using ELISA. Results: Mean IGF-I level in GCF was higher in the test group than control. Mean IGF-I level in plasma and IGFBP-3 level in GCF and plasma in control group were higher than that of the test group. However, there was no statistical significance (p > 0.05). The mean level of IGF-I and IGFBP-3 in GCF was lower than those in plasma. Mean IGF-I level in plasma showed a negative correlation with PD and CAL (p < 0.05) in both groups. The levels of IGF-I and IGFBP-3 in GCF seemed to be negatively correlated with BOP in the test group (p < 0.05). Conclusions: The difference in the level of IGF-I and IGFBP-3 between AgP and healthy subjects was not significant. Further studies that explain the mechanism of the protective role of IGF-I with more samples are needed.

Keywords: aggressive periodontitis, pathogenesis, insulin-like growth factor, insulin-like growth factor binding protein

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Authors: Emil F. Khisamutdinov

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Development of functional materials undergoing structural transformations in response to an external stimulus such as environmental changes (pH, temperature, etc.), the presence of particular proteins, or short oligonucleotides are of great interest for a variety of applications ranging from medicine to electronics. The dynamic operations of most nucleic acid (NA) devices, including circuits, nano-machines, and biosensors, rely on networks of NA strand displacement processes in which an external or stimulus strand displaces a target strand from a DNA or RNA duplex. The rate of strand displacement can be greatly increased by the use of “toeholds,” single-stranded regions of the target complex to which the invading strand can bind to initiate the reaction, forming additional base pairs that provide a thermodynamic driving force for transformation. Herein, we developed a highly robust nanoparticle shape transition, sequentially transforming DNA polygons from one shape to another using the toehold-mediated DNA strand displacement technique. The shape transformation was confirmed by agarose gel electrophoresis and atomic force microscopy. Furthermore, we demonstrate that our approach is applicable for RNA shape transformation from triangle to square, which can be detected by fluorescence emission from malachite green binding RNA aptamer. Using gel-shift and fluorescence assays, we demonstrated efficient transformation occurs at isothermal conditions (37°C) that can be implemented within living cells as reporter molecules. This work is intended to provide a simple, cost-effective, and straightforward model for the development of biosensors and regulatory devices in nucleic acid nanotechnology.

Keywords: RNA nanotechnology, bionanotechnology, toehold mediated DNA switch, RNA split fluorogenic aptamers

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Authors: Monish Sengupta, Benjamin Heydecker, Daniel Woodland

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New train control systems rely mainly on Automatic Train Protection (ATP) and Automatic Train Operation (ATO) dynamically to control the speed and hence performance. The ATP and the ATO form the vital element within the CBTC (Communication Based Train Control) and within the ERTMS (European Rail Traffic Management System) system architectures. Reliable and accurate measurement of train location, speed and acceleration are vital to the operation of train control systems. In the past, all CBTC and ERTMS system have deployed a balise or equivalent to correct the uncertainty element of the train location. Typically a CBTC train is allowed to miss only one balise on the track, after which the Automatic Train Protection (ATP) system applies emergency brake to halt the service. This is because the location uncertainty, which grows within the train control system, cannot tolerate missing more than one balise. Balises contribute a significant amount towards wayside maintenance and studies have shown that balises on the track also forms a constraint for future track layout change and change in speed profile.This paper investigates the causes of the location uncertainty that is currently experienced and considers whether it is possible to identify an effective filter to ascertain, in conjunction with appropriate sensors, more accurate speed, distance and location for a CBTC driven train without the need of any external balises. An appropriate sensor fusion algorithm and intelligent sensor selection methodology will be deployed to ascertain the railway location and speed measurement at its highest precision. Similar techniques are already in use in aviation, satellite, submarine and other navigation systems. Developing a model for the speed control and the use of Kalman filter is a key element in this research. This paper will summarize the research undertaken and its significant findings, highlighting the potential for introducing alternative approaches to train positioning that would enable removal of all trackside location correction balises, leading to huge reduction in maintenances and more flexibility in future track design.

Keywords: ERTMS, CBTC, ATP, ATO

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Authors: Mark-Adam Kellerman

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Proteins are capable of exploring vast mutational spaces. This makes it difficult for protein engineers to devise rational methods to improve stability and function via mutagenesis. Deciding which residues to mutate requires knowledge of the characteristics they elicit. We probed the characteristics of residues in granulocyte-colony stimulating factor (G-CSF) using a thermal melt (from 295K to 323K) to denature it in a 700 MHz Bruker spectrometer. These characteristics included dynamics, micro-environmental changes experienced/ induced during denaturing and structure-function relationships. 15N-1H HSQC experiments were performed at 2K increments along with this thermal melt. We observed that dynamic residues that also undergo a lot of change in their microenvironment were predominantly in unstructured regions. Moreover, we were able to identify four residues (G4, A6, T133 and Q134) that we class as high priority targets for mutagenesis, given that they all appear in both the top 10% of measures for environmental changes and dynamics (∑Δ and ∆PI). We were also able to probe these NMR observables and combine them with molecular dynamics (MD) to elucidate what appears to be an opening motion of G-CSFs binding site III. V48 appears to be pivotal to this opening motion, which also seemingly distorts the loop region between helices A and B. This observation is in agreement with previous findings that the conformation of this loop region becomes altered in an aggregation-prone state of G-CSF. Hence, we present here an approach to profile the characteristics of residues in order to highlight their potential as rational mutagenesis targets and their roles in important conformational changes. These findings present not only an opportunity to effectively make biobetters, but also open up the possibility to further understand epistasis and machine learn residue behaviours.

Keywords: protein engineering, rational mutagenesis, NMR, molecular dynamics

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Authors: Mahmoud A. Amin, Essam Taman, Ahmed Omran, Mahmoud Shawky, Ahmed Mekawy, Abdallah M. Kotkat, Saber Younes, Nehad N. Ghonemy, Amin Saad, Ezz-Aleslam, Abdullah M. Elosh

Abstract:

Introduction: TMJ is a one-of-a-kind, complicated synovial joint that helps with masticatory function by allowing the mandible to open and close the mouth. True ankylosis is a situation in which condylar movement is limited by a mechanical defect in the joint, whereas false ankylosis is a condition in which there is a restriction in mandibular movement due to muscular spasm myositis ossificans, and coronoid process hyperplasia. Ankylosis is characterized by the inability to open the mouth due to fusion of the TMJ condyle to the base of the skull as a result of trauma, infection, or systemic diseases such as rheumatoid arthritis (the most common) and psoraisis. Ankylosis causes facial asymmetry and affects the patient psychologically as well as speech, difficult mastication, poor oral hygiene, malocclusion, and other factors. TMJ is a technically challenging joint; hence TMJ ankylosis management is complicated. Case presentation: this case is a male patient 25 years old reported to our maxillofacial clinic in Damietta faculty of medicine, Al-Azhar University with the inability to open the mouth at all, with a history of difficulty of mouth breathing and eating foods, there was a history of falling from height at 2006, and the patient underwent corrective surgery before with no improvement because the ankylosis was relapsed short period after the previous operations with that done out of our hospital inter-incisor distant ZERO so, this condition need mandatory management. Clinical examination and radiological investigations were done after complete approval from the patient and his brother; tracheostomy was done for our patient before the operation. The patient entered the operation in our hospital and drastic improvement in mouth opening was noticed, helping to restore the physical psychological health of the patient.

Keywords: temporomandibular joint, TMJ, Ankylosis, mouth opening, physiotherapy, condylar plate

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Authors: F. Deng, R. Sanchez, A. Beltran, S. Maestre

Abstract:

The high nutritional value associated with seafood is related to the presence of essential trace elements. Moreover, seafood is considered an important source of energy, proteins, and long-chain polyunsaturated fatty acids. Generally, seafood is consumed cooked. Consequently, the nutritional value could be degraded. Seafood, such as fish, shellfish, and seaweed, could be considered as one of the main iodine sources. The deficient or excessive consumption of iodine could cause dysfunction and pathologies related to the thyroid gland. The main objective of this work is to evaluated iodine stability in hake (Merluccius) undergone different culinary techniques. The culinary process considered were: boiling, steaming, microwave cooking, baking, cooking en papillote (twisted cover with the shape of a sweet wrapper) and coating with a batter of flour and deep-frying. The determination of iodine was carried by Inductively Coupled Plasma Mass Spectrometry (ICP-MS). Regarding sample handling strategies, liquid-liquid extraction has demonstrated to be a powerful pre-concentration and clean-up approach for trace metal analysis by ICP techniques. Extraction with tetramethylammonium hydroxide (TMAH reagent) was used as a sample preparation method in this work. Based on the results, it can be concluded that the stability of iodine was degraded with the cooking processes. The major degradation was observed for the boiling and microwave cooking processes. The content of iodine in hake decreased up to 60% and 52%, respectively. However, if the boiling cooking liquid is preserved, this loss that has been generated during cooking is reduced. Only when the fish was cooked by following the cooking en papillote process the iodine content was preserved.

Keywords: cooking process, ICP-MS, iodine, hake

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Authors: Samaneh Farokhirad, Fatemeh Ahmadpoor

Abstract:

Biological membranes are constantly in contact with various filamentous soft nanostructures that either reside on their surface or are being transported between the cell and its environment. In particular, viral infections are determined by the interaction of viruses (such as filovirus) with cell membranes, membrane protein organization (such as cytoskeletal proteins and actin filament bundles) has been proposed to influence the mechanical properties of lipid membranes, and the adhesion of filamentous nanoparticles influence their delivery yield into target cells or tissues. The goal of this research is to integrate the rapidly increasing but still fragmented experimental observations on the adhesion and self-assembly of nanofilaments (including filoviruses, actin filaments, as well as natural and synthetic nanofilaments) on cell membranes into a general, rigorous, and unified knowledge framework. The global outbreak of the coronavirus disease in 2020, which has persisted for over three years, highlights the crucial role that nanofilamentbased delivery systems play in human health. This work will unravel the role of a unique property of all cell membranes, namely flexoelectricity, and the significance of nanofilaments’ flexibility in the adhesion and self-assembly of nanofilaments on cell membranes. This will be achieved utilizing a set of continuum mechanics, statistical mechanics, and molecular dynamics and Monte Carlo simulations. The findings will help address the societal needs to understand biophysical principles that govern the attachment of filoviruses and flexible nanofilaments onto the living cells and provide guidance on the development of nanofilament-based vaccines for a range of diseases, including infectious diseases and cancer.

Keywords: virus nanofilaments, cell mechanics, computational biophysics, statistical mechanics

Procedia PDF Downloads 93

Authors: Arefeh Jafarian, Mohammad Taghikani, Saied Abroun, Amir Allahverdi, Masoud Soleimani

Abstract:

Introduction: The miRNAs have key roles in control of pancreatic islet development and insulin secretion. In this regards, current study investigated the pancreatic differentiation of human bone marrow mesenchymal stem cells (hBM-MSCs) by up-regulation of miR-375 and down-regulation of miR-9 by lentiviruses containing miR-375 and anti-miR-9. Findings: After 21 days of induction, islet-like clusters containing insulin producing cells (IPCs) were confirmed by dithizone (DTZ) staining. The IPCs and β cell specific related genes and proteins were detected using qRT-PCR and immunofluorescence on days 7, 14 and 21 of differentiation. Glucose challenge test was performed at different concentrations of glucose as well as extracellular and intracellular insulin and C-peptide were assayed using ELISA kit. In derived IPCs by miR-375 alone are capable to express insulin and other endocrine specific transcription factors, the cells lack the machinery to respond to glucose. The differentiated hMSCs by miR-375 and anti-miR-9 lentiviruses could secrete insulin and c-peptide in a glucose-regulated manner. Conclusion: It was found that over-expression of miR-375 led to a reduction in levels of Mtpn protein in derived IPCs, while treatment with anti-miR-9 following miR-375 over-expression had synergistic effects on MSCs differentiation and insulin secretion in a glucose-regulated manner. The researchers reported that silencing of miR-9 increased OC-2 protein in IPCs that may contribute to the observed glucose-regulated insulin secretion. These findings highlight miRNAs functions in stem cells differentiation and suggest that they could be used as therapeutic tools for gene-based therapy in diabetes mellitus.

Keywords: diabetes, differentiation, MSCs, insulin producing cells, miR-375, miR-9

Procedia PDF Downloads 317

Authors: A. M. Malagon Escandon, J. A. Arenas Alatorre, C. P. Chaires Rosas, N. A. Vazquez Torres, B. Hernandez Tellez, G. Pinon Zarate, M. Herrera Enriquez, A. E. Castell Rodriguez

Abstract:

The current approach to the treatment of bone defects involves the use of scaffolds that provide a biological and mechanically stable niche to favor tissue repair. Despite the significant progress in the field of bone tissue engineering, several main problems associated are attributed to giving a low biodegradation degree, does not promote osseointegration and regeneration, if the bone is not healing as well as expected or fails to heal, will not be given a proper ossification or new bone formation. The actual approaches of bone tissue regeneration are directed to the use of decellularized native extracellular matrices, which are able of retain their own architecture, mechanic properties, biodegradability and promote new bone formation because they are capable of conserving proteins and other factors that are founded in physiological concentrations. Therefore, we propose an extracellular matrix-based bioscaffolds derived from bovine cancellous bone, which is processed by decellularization, demineralization, and hydrolysis of the collagen protein, these protocols have been successfully carried out in other organs and tissues; the effectiveness of its biosafety has also been previously evaluated in vivo and Food and Drug Administration (FDA) approved. In the specific case of bone, a more complex treatment is needed in comparison with other organs and tissues because is necessary demineralization and collagen denaturalization. The present work was made in order to obtain a temporal scaffold that succeed in degradation in an inversely proportional way to the synthesis of extracellular matrix and the maturation of the bone by the cells of the host.

Keywords: bioactive, biodegradable, bone, extracellular matrix-based bioscaffolds, stem cells, tissue engineering

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Authors: Mayank, Navneet Kaur, Narinder Singh

Abstract:

The V599 mutations in the BRAF protein are extremely oncogenic, responsible for countless of malignant conditions. Along with wild type, V599E, V599D, and V599R are the important mutated variants of the BRAF proteins. The BRAF inhibitory anticancer agents are continuously developing, and sorafenib is a BRAF inhibitor that is under clinical use. The crystal structure of sorafenib bounded to wild type, and V599 is known, showing a similar interaction pattern in both the case. The mutated 599th residue, in both the case, is also found not interacting directly with the co-crystallized sorafenib molecule. However, the IC50 value of sorafenib was found extremely different in both the case, i.e., 22 nmol/L for wild and 38 nmol/L for V599E protein. Molecular docking study and MMGBSA binding energy results also revealed a significant difference in the binding pattern of sorafenib in both the case. Therefore, to explore the role of distinctively situated 599th residue, we have further conducted comprehensive computational studies. The molecular dynamics simulation, residue interaction network (RIN) analysis, and residue correlation study results revealed the importance of the 599th residue on the therapeutic outcome and overall dynamic of the BRAF protein. Therefore, although the position of 599th residue is very much distinctive from the ligand-binding cavity of BRAF, still it has exceptional control over the overall functional outcome of the protein. The insight obtained here may seem extremely important and guide us while designing ideal BRAF inhibitory anticancer molecules.

Keywords: BRAF, oncogenic, sorafenib, computational studies

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Authors: Michael A. Sprayberry, Vincent C. Paquit

Abstract:

Process parameter optimization in metal powder bed electron beam melting (MPBEBM) is crucial to ensure the technology's repeatability, control, and industry-continued adoption. Despite continued efforts to address the challenges via the traditional design of experiments and process mapping techniques, there needs to be more successful in an on-the-fly optimization framework that can be adapted to MPBEBM systems. Additionally, data-intensive physics-based modeling and simulation methods are difficult to support by a metal AM alloy or system due to cost restrictions. To mitigate the challenge of resource-intensive experiments and models, this paper introduces a Multi-Objective Reinforcement Learning (MORL) methodology defined as an optimization problem for MPBEBM. An off-policy MORL framework based on policy gradient is proposed to discover optimal sets of beam power (P) – beam velocity (v) combinations to maintain a steady-state melt pool depth and phase transformation. For this, an experimentally validated Eagar-Tsai melt pool model is used to simulate the MPBEBM environment, where the beam acts as the agent across the P – v space to maximize returns for the uncertain powder bed environment producing a melt pool and phase transformation closer to the optimum. The culmination of the training process yields a set of process parameters {power, speed, hatch spacing, layer depth, and preheat} where the state (P,v) with the highest returns corresponds to a refined process parameter mapping. The resultant objects and mapping of returns to the P-v space show convergence with experimental observations. The framework, therefore, provides a model-free multi-objective approach to discovery without the need for trial-and-error experiments.

Keywords: additive manufacturing, metal powder bed fusion, reinforcement learning, process parameter optimization

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Authors: Benchouala Amira, Bojic Clement, Poupin Pascal, Cossu Leguille-carole

Abstract:

The objective of this study is to evaluate in vitro the cytotoxic and androgenic potential of several antifungal molecules (amphotericin B, econazole, ketoconazole and miconazole) on MDA-Kb2 cell lines. This biological model is an effective tool for the detection of endocrine disruptors because it responds well to the main agonist of the androgen receptor (testosterone) and also to an antagonist: flutamide. The cytotoxicity of each chemical compound tested was measured using an MTT assay (tetrazolium salt, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) which measures the activity of the reductase function of mitochondrial succinate dehydrogenase enzymes of cultured cells. This complementary cytotoxicity test is essential to ensure that the effects of reduction in luminescence intensity observed during androgenic tests are only attributable to the anti-androgenic action of the compounds tested and not to their possible cytotoxic properties. Tests of the androgenic activity of antifungals show that these compounds do not have the capacity to induce transcription of the luciferase gene. These compounds do not exert an androgenic effect on MDA-Kb2 cells in culture for the environmental concentrations tested. The addition of flutamide for the same tested concentrations of antifungal molecules reduces the luminescence induced by amphotericin B, econazole and miconazole, which is explained by a strong interaction of these molecules with flutamide which may have a greater toxic effect than when tested alone. The cytotoxicity test shows that econazole and ketoconazole can cause cell death at certain concentrations tested. This cell mortality is perhaps induced by a direct or indirect action on deoxyribonucleic acid (DNA), ribonucleic acid (RNA) or proteins necessary for cell division.

Keywords: cytotoxicity, androgenic potential, antifungals, MDA-Kb2

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Authors: Ahmad Ali Shahid, M Shakil Shaukat

Abstract:

Agriculture is the backbone of economy of Pakistan and Cotton is the major agricultural export and supreme source of raw fiber for our textile industry. To combat against the developing resistance in the target insects and combating these challenges wholesomely, a novel combination of pyramided/stacked genes was conceptualized and later realized, through the means of biotechnology i.e., transformation of three genes namely, Cry1Ac, Cry2A, and EPSP synthase (glyphosate tolerant) genes in the locally cultivated cotton variety. The progenies of the transformed plants were successfully raised and screened under the tunnel conditions for two generations and the present study focused on the screening of plants which were confirmed for containing all of these three genes and their expressions. Initially, the screening was done through glyphosate spray assay and the plants which were healthy and showed no damage on leaves were selected after 07 days of spray. In the laboratory, the DNA of these plants were isolated and subjected to amplification of the three genes. Thus, seventeen out of twenty were confirmed positive for Cry1Ac gene and ten out of twenty were positive for Cry2A gene and all twenty were positive for presence of EPSP synthase gene. Then, the ten plant samples which were confirmed with presence of all three genes were subjected to expression analysis of these proteins through ELISA. The results showed that eight out of ten plants were actively expressing the three transgenes. Real-time PCR was also done to quantify the expression levels of the EPSP synthase gene. Finally, eight plants were confirmed for the presence and active expression of all three genes in T3 generation of the triple gene transformed cotton. These plants may be subjected to T4 generation to develop a new stable variety in due course of time.

Keywords: agriculture, cotton, transformation, cry genes, ELISA, PCR

Procedia PDF Downloads 394

Authors: Ibrahim Iddrisu, Joseph Ayariga, Junhuan Xu, Ayomide Adebanjo, Boakai K. Robertson, Michelle Samuel-Foo, Olufemi Ajayi

Abstract:

The rise of antimicrobial resistance is a global public health crisis that threatens the effective control and prevention of infections. Due to the emergence of pan drug-resistant bacteria, most antibiotics have lost their efficacy. Bacteriophages or their components are known to target bacterial cell walls, cell membranes, and lipopolysaccharides (LPS) and hydrolyze them. Bacteriophages, being the natural predators of pathogenic bacteria, are inevitably categorized as ‘human friends’, thus fulfilling the adage that ‘the enemy of my enemy is my friend’. Leveraging on their lethal capabilities against pathogenic bacteria, researchers are searching for more ways to overcome the current antibiotic resistance challenge. In this study, we expressed and purified epsilon 34 phage tail spike protein (E34 TSP) from the E34 TSP gene, then assessed the ability of this bacteriophage protein in the killing of two CBD-resistant strains of Salmonella spp. We also assessed the ability of the tail spike protein to cause bacteria membrane disruption and dehydrogenase depletion. We observed that the combined treatment of CBD-resistant strains of Salmonella with CBD and E34 TSP showed poor killing ability, whereas the mono treatment with E34 TSP showed considerably higher killing efficiency. This study demonstrates that the inhibition of the bacteria by E34 TSP was due in part to membrane disruption and dehydrogenase inactivation by the protein. The results of this work provide an interesting background to highlight the crucial role phage proteins such as E34 TSP could play in pathogenic bacterial control.

Keywords: cannabidiol, resistance, Salmonella, antimicrobials, phages

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Authors: Asmaa M. Fahim

Abstract:

In this elucidation, we synthesized different heterocyclic compounds attached to Benzene sulfonamide moiety via (E)-N-(4-(3-(4-bromophenyl)acryloyl)phenyl)-4-methyl benzene sulfonamide which is obtained from Nucleophilic substitution reaction between 4-methylbenzene sulfonyl chloride and 1-(4-aminophenyl)ethan-1-one in pyridine to get N-(4-acetyl phenyl)-4-methyl benzenesulfonamide which reacted 4-bromobenzal dehyde undergoes aldol condensation in NaOH to afford the corresponding chalchone 4. Moreover, the reactivity of chalchone 4 showed several active methylene derivatives utilized the pressurized microwave irradiation as a green energy resource. Chalcone 4 was allowed to react with ethyl cyanoacetate and acetylacetone, respectively, at 70 °C with pressure under microwave reaction condition to afford the 5-cyano-6-oxo-1,2,5,6-tetrahydropyridin-2-yl)-4-methylbenzenesulfonamide 6 and N-(4'-acetyl-4''-bromo-5'-oxo-2',3',4',5'-tetrahydro-[1,1':3',1''-terphenyl]-4-yl)-4-methylbenzenesulfonamide 8 derivatives. Moreover, the reactivity of this sulphonamide chalchone with NH2NH2 in EtOH and acetic acid, which gave 2,5-dihydro-1H-imidazol-4-yl)-4-methyl benzenesulfonamide, 1H-pyrazol-3-yl)-4-methyl and reactivity with NH2OH.HCl gave isoxazol-3-yl)-4-methylbenzenesulfonamide derivatives. The synthesized compounds were screened for their ADME properties and directed to antitumor activity on HepG2 hepatocellular carcinoma and MCF-7 breast cancer and exhibited excellent behavior against standard drugs; these results were confirmed through molecular simulations with different proteins. Additionally, the Density Functional Theory analysis of optimized structures investigated their physical descriptors, FMO, ESP and MEP, which correlated with biological evaluation.

Keywords: synthesis, green chemistry, antitumor activity, DFT study

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Authors: H. Aldewachi, M. Hines, M. McCulloch, N. Woodroofe, P. Gardiner

Abstract:

DPP-IV is an enzyme whose expression is affected in a variety of diseases, therefore, has been identified as possible diagnostic or prognostic marker for various tumours, immunological, inflammatory, neuroendocrine, and viral diseases. Recently, DPP-IV enzyme has been identified as a novel target for type II diabetes treatment where the enzyme is involved. There is, therefore, a need to develop sensitive and specific methods that can be easily deployed for the screening of the enzyme either as a tool for drug screening or disease marker in biological samples. A variety of assays have been introduced for the determination of DPP-IV enzyme activity using chromogenic and fluorogenic substrates, nevertheless these assays either lack the required sensitivity especially in inhibited enzyme samples or displays low water solubility implying difficulty for use in vivo samples in addition to labour and time-consuming sample preparation. In this study, novel strategies based on exploiting the high extinction coefficient of gold nanoparticles (GNPs) are investigated in order to develop fast, specific and reliable enzymatic assay by investigating synthetic peptide sequences containing a DPP IV cleavage site and coupling them to GNPs. The DPP IV could be detected by colorimetric response of peptide capped GNPs (P-GNPS) that could be monitored by a UV-visible spectrophotometer or even naked eyes, and the detection limit could reach 0.01 unit/ml. The P-GNPs, when subjected to DPP IV, showed excellent selectivity compared to other proteins (thrombin and human serum albumin) , which led to prominent colour change. This provided a simple and effective colorimetric sensor for on-site and real-time detection of DPP IV.

Keywords: gold nanoparticles, synthetic peptides, colorimetric detection, DPP-IV enzyme

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