Search results for: bioactive peptides

Authors: A. Aslihan Gokaltun, Ali Tuncel

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The unique properties of capillary electrochromatography (CEC) such as high performance, high selectivity, low consumption of both reagents and analytes ensure this technique an attractive one for the separation of biomolecules including nucleosides and nucleotides, peptides, proteins, carbohydrates. Monoliths have become a well-established separation media for CEC in the format that can be compared to a single large 'particle' that does not include interparticular voids. Convective flow through the pores of monolith significantly accelerates the rate of mass transfer and enables a substantial increase in the speed of the separation. In this work, we propose a new approach for the preparation of cationic monolithic stationary phase for capillary electrochromatography. Instead of utilizing a charge bearing monomer during polymerization, the desired charge-bearing group is generated on the capillary monolith after polymerization by using the reactive moiety of the monolithic support via one-pot, simple reaction. Optimized monolithic column compensates the disadvantages of frequently used reversed phases, which are difficult for separation of polar solutes. Rapid separation and high column efficiencies are achieved for the separation of neutral analytes, nucleic acid bases and nucleosides in reversed phase mode. Capillary monolith showed satisfactory hydrodynamic permeability and mechanical stability with relative standard deviation (RSD) values below 2 %. A new promising, reactive support that has a 'ligand selection flexibility' due to its reactive functionality represent a new family of separation media for CEC.

Keywords: biomolecules, capillary electrochromatography, cationic monolith, neutral analytes

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Authors: D. J. Kalita

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Cancer is one of the prime causes of early death worldwide. Mutation of the gene involve in DNA repair and damage, like BRCA2 (Breast cancer gene two) genes, can be detected efficiently by PCR-RFLP to early breast cancer diagnosis and adopt the suitable method of treatment. Host Defense Peptide can be used as blueprint for the design and synthesis of novel anticancer drugs to avoid the side effect of conventional chemotherapy and chemo resistance. The change at nucleotide position 392 of a -› c in the cancer sample of dog mammary tumour at BRCA2 (exon 7) gene lead the creation of a new restriction site for SsiI restriction enzyme. This SNP may be a marker for detection of canine mammary tumour. Support vector machine (SVM) algorithm was used to design and predict the anticancer peptide from the mature functional peptide. MTT assay of MCF-7 cell line after 48 hours of post treatment showed an increase in the number of rounded cells when compared with untreated control cells. The ability of the synthesized peptide to induce apoptosis in MCF-7 cells was further investigated by staining the cells with the fluorescent dye Hoechst stain solution, which allows the evaluation of the nuclear morphology. Numerous cells with dense, pyknotic nuclei (the brighter fluorescence) were observed in treated but not in control MCF-7 cells when viewed using an inverted phase-contrast microscope. Thus, PCR-RFLP is one of the attractive approach for early diagnosis, and synthetic cationic peptide can be used for the treatment of canine mammary tumour.

Keywords: cancer, cationic peptide, host defense peptides, Breast cancer genes

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Authors: Vu Thu Trang, Lam Xuan Thanh, Samira Sarter, Tomoko Shimamura, Hiroaki Takeuchi　　

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Antimicrobial activities of aminoreductone (AR), a product formed in the initial stage of Maillard reaction, were screened against pathogenic bacteria. A significant growth inhibition of AR against all 7 isolates (Staphylococcus aureus ATCC® 25923™, Salmonella Typhimurium ATCC® 14028™, Bacillus cereus ATCC® 13061™, Bacillus subtilis ATCC® 11774™, Escherichia coli ATCC® 25922™, Enterococcus faecalis ATCC® 29212™, Listeria innocua ATCC® 33090™) were observed by the standard disc diffusion methods. The inhibition zone for each isolate by AR (2.5 mg) ranged from 15±0 mm to 28.3±0.4 mm in diameter. The minimum inhibitory concentration (MIC) of AR ranging from 20 mM to 26 mM was proven in the seven isolates tested. AR also showed the similar effect of growth inhibition in comparison with antibiotics frequently used for the treatment of infections bacteria, such as amikacin, ciprofloxacin, meropennem, and levofloxacin. The results indicated that foods containing AR are valuable sources of bioactive compounds towards pathogenic bacteria.

Keywords: pathogenic bacteria, aminoreductone, Maillard reaction, antimicrobial activity

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Authors: Prachi Singh

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This paper presents a low-cost, eco-friendly and reproducible microbe mediated biosynthesis of TiO₂ nanoparticles. TiO₂ nanoparticles synthesized using the bacterium, Bacillus subtilis, from titanium as a precursor, were confirmed by TEM analysis. The morphological characteristics state spherical shape, with the size of individual or aggregate nanoparticles, around 30-40 nm. Microbial resistance represents a challenge for the scientific community to develop new bioactive compounds. Here, the antibacterial effect of TiO₂ nanoparticles on Escherichia coli was investigated, which was confirmed by CFU (Colony-forming unit). Further, growth curve study of E. coli Hb101 in the presence and absence of TiO₂ nanoparticles was done. Optical density decrease was observed with the increase in the concentration of TiO₂. It could be attributed to the inactivation of cellular enzymes and DNA by binding to electron-donating groups such as carboxylates, amides, indoles, hydroxyls, thiols, etc. which cause little pores in bacterial cell walls, leading to increased permeability and cell death. This justifies that TiO₂ nanoparticles have efficient antibacterial effect and have potential to be used as an antibacterial agent for different purposes.

Keywords: antibacterial effect, CFU, Escherichia coli Hb101, growth curve, TEM, TiO2 nanoparticle, Toxicity, UV-Vis

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Authors: Christophe Gonçalves, Raquel P. F. Guiné, Daniela Teixeira, Fernando J. Gonçalves

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Blueberries are widely valued for their high content in phenolic compounds with antioxidant activity, and hence beneficial for the human health. In this way, a study was done to determine the phenolic composition (total phenols, anthocyanins and tannins) and antioxidant activity of blueberries from three cultivars (Duke, Bluecrop, and Ozarblue) grown in two different Portuguese farms. Initially two successive extractions were done with methanol followed by two extractions with aqueous acetone solutions. These extracts obtained were then used to evaluate the amount of phenolic compounds and the antioxidant activity. The total phenols were observed to vary from 4.9 to 8.2 mg GAE/g fresh weight, with anthocyanin’s contents in the range 1.5-2.8 mg EMv3G/g and tannins contents in the range 1.5- 3.8 mg/g. The results for antioxidant activity ranged from 9.3 to 23.2 mol TE/g, and from 24.7 to 53.4 mol TE/g, when measured, respectively, by DPPH and ABTS methods. In conclusion it was observed that, in general, the cultivar had a visible effect on the phenols present, and furthermore, the geographical origin showed relevance either in the phenols contents or the antioxidant activity.

Keywords: anthocyanins, antioxidant activity, blueberry cultivar, geographical origin, phenolic compounds

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Authors: Serier Bouchenak NORA, Bouguerni ABDELMADJID

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Algeria is a Mediterranean country which provides an ideal habitat for a wide range of species of medicinal plants. The objective of this current work is to extract the gel contained in the leaves of Aloe vera in order to formulate an oral gel as a prebiotic and see its effects on probiotics (lactic and pseudo lactic bacteria and bifido bacterium). Aloe vera polysaccharid extract is a matrix mainly composed of non-digestible oligosaccharids or slow-fermentation polysaccharids, as this produces a lower pH. The behavior of Aloe vera during in vitro fermentation of the colon was similar to that of lactulose, indicating the possibility of using Aloe vera and its polysaccharids extracts as a prebiotic. The microbiological control of the two kinds of bacteria (bifidobacteria and staphylococci) has demonstrated the gel capacity to stimulate them by these bioactive compounds. The generation time of Bifidobacteria in fermented milk with added prebiotic Aloe vera gel is 80.408 min with a µ growth rate equal to 0.012 min -1. The doubling time is 61.459 min with a growth rate µ equal to 0.016 min -1 for the Streptococcus sp. species.

Keywords: aloe vera, probiotics, prebiotics, growth rate, bifidobacteria

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Authors: K. Fellah, H. Benmehdi, A. Amrouche, H. Malainine, F. Memmou, H. Dalile, W. Siata

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This work aims to evaluate the antioxidant of flavonoids extracted from the leaves of Prunus persica L. A phytochemical screening allowed us to highlight the different phytochemicals present in the leaves of the studied plant. The selective extraction of flavonoids gave yields of 0.71, 1.5, and 4.8% for the fractions ethyl ether, ethyl acetate and n- butanol, respectively. The reading of the antioxidant activity of different extracts of flavonoids by HPLTC method revealed positive reaction (yellow spots) on the TLC plates sprayed with DPPH. Using the DPPH method, the fractions of flavonoids (bunanol, ethyl acetate and Diethyl ether) showed a potent scavenging activity with IC50 = 0.22; 0.27 and 0.76 mg / ml, respectively. Furthermore, our findings revealed the extracts under study exhibited higher reducing potential which depends upon extract concentration. These results obtained from this investigation confirm that the Prunus persica remains a major resource of bioactive molecules.

Keywords: Prunus persica L., phytochemical study, flavonoids, antioxidant activity, TLC bioautographic, FRAP, DPPH

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Authors: Shalini Singh, Sanjay K. Srivastava, Govind, Mukhtar. A. Khan, P. K. Singh

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Nanotechnology is revolutionizing the development of biosensors. Nanomaterials and nanofabrication technologies are increasingly being used to design novel biosensors. Sensitivity and other attributes of biosensors can be improved by using nanomaterials with unique chemical, physical, and mechanical properties in their construction. Silicon is a promising biomaterial that is non-toxic and biodegradable and can be exploited in chemical and biological sensing. Present study demonstrated the streptavidin–biotin interaction on silicon surfaces with different topographies such as flat and nanostructured silicon (nanowires) surfaces. Silicon nanowires with wide range of surface to volume ratio were prepared by electrochemical etching of silicon wafer. The large specific surface of silicon nanowires can be chemically modified to link different molecular probes (DNA strands, enzymes, proteins and so on), which recognize the target analytes, in order to enhance the selectivity and specificity of the sensor device. The interaction of streptavidin with biotin was carried out on 3-aminopropyltriethoxysilane (APTS) functionalized silicon surfaces. Fourier Transform Infrared Spectroscopy (FTIR) and X-ray Photoelectron Spectroscopy (XPS) studies have been performed to characterize the surface characteristics to ensure the protein attachment. Silicon nanowires showed the enhance protein attachment, as compared to flat silicon surface due to its large surface area and good molecular penetration to its surface. The methodology developed herein could be generalized to a wide range of protein-ligand interactions, since it is relatively easy to conjugate biotin with diverse biomolecules such as antibodies, enzymes, peptides, and nucleotides.

Keywords: FTIR, silicon nanowires, streptavidin-biotin, XPS

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Authors: Tatjana Kadifkova Panovska, Svetlana Kulevanova, Blagica Jovanova

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Biological systems possess the ability to neutralize the excess of reactive oxygen species (ROS) and to protect cells from destructive alterations. However, many pathological conditions (cardiovascular diseases, autoimmune disorders, cancer) are associated with inflammatory processes that generate an excessive amount of reactive oxygen species (ROS) that shift the balance between endogenous antioxidant systems and free oxygen radicals in favor of the latter, leading to oxidative stress. Therefore, an additional source of natural compounds with antioxidant properties that will reduce the amount of ROS in cells is much needed despite their broad utilization; many plant species remain largely unexplored. Therefore, the purpose of the present study is to investigate the antioxidant activity of twenty-five selected medicinal and aromatic plant species. The antioxidant activity of the ethanol extracts was evaluated with in vitro assays: 2,2’-diphenyl-1-pycryl-hydrazyl (DPPH), ferric reducing antioxidant power (FRAP), non-site-specific- (NSSOH) and site-specific hydroxyl radical-2-deoxy-D-ribose degradation (SSOH) assays. The Folin-Ciocalteu method and AlCl3 method were performed to determine total phenolic content (TPC) and total flavonoid content (TFC). All examined plant extracts manifested antioxidant activity to a different extent. Cinnamomum verum J.Presl bark and Ocimum basilicum L. Herba demonstrated strong radical scavenging activity and reducing power with the DPPH and FRAP assay, respectively. Additionally, significant hydroxyl scavenging potential and metal chelating properties were observed using the NSSOH and SSOH assays. Furthermore, significant variations were determined in the total polyphenolic content (TPC) and total flavonoid content (TFC), with Cinnamomum verum and Ocimum basilicum showing the highest amount of total polyphenols. The considerably strong radical scavenging activity, hydroxyl scavenging potential and reducing power for the species mentioned above suggest of a presence of highly bioactive phytochemical compounds, predominantly polyphenols. Since flavonoids are the most abundant group of polyphenols that possess a large number of available reactive OH groups in their structure, it is considered that they are the main contributors to the radical scavenging properties of the examined plant extracts. This observation is supported by the positive correlation between the radical scavenging activity and the total polyphenolic and flavonoid content obtained in the current research. The observations from the current research nominate Cinnamomum verum bark and Ocimum basilicum herba as potential sources of bioactive compounds that could be utilized as antioxidative additives in the food and pharmaceutical industries. Moreover, the present study will help the researchers as basic data for future research in exploiting the hidden potential of these important plants that have not been explored so far.

Keywords: ethanol extracts, radical scavenging activity, reducing power, total polyphenols.

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Authors: Sibel Daglilar, Isil Kerti, Murat Karagoz, Fatih Dumludag, Oguzhan Gunduz, Faik Nuzhet Oktar

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Hydroxyapatite (HA) materials have common use in bone repairing due to its ability to accelerate the bone growth around the implant. In spite of being a biocompatible and bioactive material, HA has a limited usage as an implant material because of its weak mechanical properties. HA based composites are required to improve the strength and toughness properties of the implant materials without compromising of biocompatibility. The excellent mechanical properties and higher biocompatibilities are expected from each of biomedical composites. In this study, HA composites were synthesized by using bovine bone reinforced doped with different amount of (wt.%) Li2O. The pressed pellets were sintered at various sintering temperatures between 1000ºC and 1300°C, and mechanical, electrical properties of the obtained products were characterized. In addition to that, in vitro stimulated body fluid (SBF) tests for these samples were conducted. The most suitable composite composition for biomedical applications was discussed among the composites studied.

Keywords: biocomposites, sintering temperature, biocompatibility, electrical property, conductivity, mechanical property

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Authors: Kieren Luellman, Makenzi Rockwell, Eduardo Callegari, Nichole Haag, Chun Wu

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Multiple sclerosis (MS) is an autoimmune disorder that damages the myelin sheath of neurons in the central nervous system. The presence of Acinetobacter bacteria and anti-Acinetobacter antibodies in MS patients has led to the hypothesis that the bacteria may contribute to MS pathogenesis. In this study, the protein sequences of Acinetobacter baumannii were compared to five peptides from three mammalian myelin proteins, i.e., Proteolipid Protein (PLP): PLP 139-151, PLP 178-191, Myelin Basic Protein (MBP): MBP 84-104 and Myelin Oligodendrocyte Glycoprotein (MOG): MOG 35-55 and MOG 92-106 respectively, known to induce experimental autoimmune encephalomyelitis (EAE), a condition similar to MS. We found 11 hits (i.e., with five or more amino acid sequence similarity) in Acinetobacter baumannii, which are identical or similar to PLP139-151, 32 hits to PLP178-191, 35 to MBP 84-104, 41 hits to MOG 35-55 and 26 hits to MOG92-106. In addition, Western blotting was used to assess possible interaction between the bacterial proteins and human anti-MBP, anti-MOG, and anti-PLP antibodies produced in rabbits, corresponding to MBP 84-104, MOG 35-55, and PLP 139-151, respectively. We found that both human Polyclonal anti-MOG antibody and anti-PLP antibody recognized a protein or more proteins of the same molecular mass of around 25 kDa. in Acinetobacter baumannii. The results suggested that this/these protein(s) might potentially serve as antigen(s) to induce anti-MOG antibody and anti-PLP antibody production in mammalian B cells. The proteomic study identified 433 hits, among which the sequence of Acinetobacter baumannii protein 491 subunit A matches a previously published enzyme Acinetobacter 3-Oxoadipate CoA-Transferase, in which a fragment of its peptide was observed to recognize MS patient serum via ELISA method. Our findings might pave the road to understanding one of the pathogenesis mechanisms of MS.

Keywords: multiple sclerosis, pathogenesis, Acinetobacter baumannii, antibody recognition

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Authors: N. A. Zainuddin, I. Norhuda, I. S. Adeib, A. N. Mustapa, S. H. Sarijo

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Recently, RESS (Rapid Expansion Supercritical Solution) method has been used by researchers to produce fine particles for pharmaceutical drug substances. Since RESS technology acknowledges a lot of benefits compare to conventional method of ginger extraction, it is suggested to use this method to explore particle formation of bioactive compound from powder ginger. The objective of this research is to produce direct solid oil particles formation from ginger rhizome which contains valuable compounds by using RESS-CO₂ process. RESS experiments were carried using extraction pressure of 3000, 4000, 5000, 6000 and 7000psi and at different extraction temperature of 40, 45, 50, 55, 60, 65 and 70°C for 40 minutes extraction time and contant flowrate (24ml/min). From the studies conducted, it was found that at extraction pressure 5000psi and temperature 40°C, the smallest particle size obtained was 2.22μm on 99 % reduction from the original size of 370μm.

Keywords: particle size, RESS, solid oil particle, supercritical carbon dioxide,

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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: Farhan Saeed

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

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

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Authors: Purwati, Sony Wibisono, Ari Sutjahjo, Askandar T. J., Fedik A. Rantam

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Diabetes mellitus type 2 (T2DM), also known as hyperglycemia, results from insulin resistance and relative insulin deficiency. Diabetes mellitus is the main cause of premature death, particularly among individuals under the age of 70 years old. Mesenchymal stem cells (MSCs) can release bioactive molecules that promote tissue repair and regeneration. Hence, in this research, we evaluated the potential of autologous adipose-derived mesenchymal stem cells (AD-MSCs) in 40 patients of phase I clinical trial in T2DM with various ages between 30-79 years. AD-MSCs are transferred through catheterization. MSCs were validated by measures of CD105+ and CD34- expression. The result showed that after AD-MSCs transplantation, blood glucose levels (fasting and 2-hour postprandial) and insulin levels were significantly decreasing. Besides that, the level of HbA1c significantly decreased after three months of AD-MSCs injection and increasing level of c-peptide after injection. Thus, we conclude that AD-MSCs injection has the potential for T2DM therapy.

Keywords: glucose, hyperglycemia, MSCs, T2DM

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Authors: Tivani Phosa Mashamba-Thompson, Mahmoud E. S. Soliman

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To date, the cause of neurodegeneration is not well understood and diseases that stem from neurodegeneration currently have no known cures. Cathepsin B (CB) enzyme is known to be involved in the production of peptide neurotransmitters and toxic peptides in neurodegenerative diseases (NDs). CA-074Me is a membrane-permeable irreversible selective cathepsin B (CB) inhibitor as confirmed by in vivo studies. Due to the lack of the crystal structure, the binding mode of CA-074Me with the human CB at molecular level has not been previously reported. The main aim of this study is to gain an insight into the binding mode of CB CA-074Me to human CB using various computational tools. Herein, molecular dynamics simulations, binding free energy calculations and per-residue energy decomposition analysis were employed to accomplish the aim of the study. Another objective was to identify novel CB inhibitors based on the structure of CA-074Me using fragment based drug design using scaffold hoping drug design approach. Results showed that two of the designed ligands (hit 1 and hit 2) were found to have better binding affinities than the prototype inhibitor, CA-074Me, by ~2-3 kcal/mol. Per-residue energy decomposition showed that amino acid residues Cys29, Gly196, His197 and Val174 contributed the most towards the binding. The Van der Waals binding forces were found to be the major component of the binding interactions. The findings of this study should assist medicinal chemist towards the design of potential irreversible CB inhibitors.

Keywords: cathepsin B, scaffold hopping, docking, molecular dynamics, binding-free energy, neurodegerative diseases

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Authors: Damandeep Kaur, O.P. Pandey, M.S. Reddy

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In the present research, bio-synthesisation of silica particles has been carried out successfully. For this purpose, agriculture waste rice husk (RH) has been utilized. Among several types of agriculture waste, RH is considered to be cost-effective and easily accessible. In the present investigation, a chemical approach has been followed to extract silica nanoparticles. X-Ray Diffraction (XRD) patterns indicated the amorphous nature of silica at lower temperature range. Silica and other mineral contents have been found using energy dispersive spectroscopy (EDS). Morphological and structural studies have been carried out with the use of Field Emission Scanning Electron Microscopy (FE-SEM) and Fourier Transform Infrared Transmission (FTIR) spectroscopy. Further, extracted silica from RH has been used for preparation of the glasses. The appearance of broad humps in XRD patterns confirmed the amorphous nature of prepared glasses. These glasses exhibited enhanced antibacterial effect against both Gram-positive and Gram-negative bacteria. The as-synthesized glass samples can be further used for physical and structural studies for drug loading applications.

Keywords: rice husk, biosynthesized silica, bioactive glasses, antibacterial studies

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Authors: Lizeth Fuentes-Mera, Vanessa Perez-Silos, Nidia K. Moncada-Saucedo, Alejandro Garcia-Ruiz, Alberto Camacho, Jorge Lara-Arias, Ivan Marino-Martinez, Victor Romero-Diaz, Adolfo Soto-Dominguez, Humberto Rodriguez-Rocha, Hang Lin, Victor Pena-Martinez

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Biphasic scaffolds in cartilage tissue engineering have been designed to influence not only the recapitulation of the osteochondral architecture but also to take advantage of the healing ability of bone to promote the implant integration with the surrounding tissue and then bone restoration and cartilage regeneration. This study reports the development and characterization of a biphasic scaffold based on the assembly of a cartilage phase constituted by fibroin biofunctionalized with bovine cartilage matrix; cellularized with differentiated pre-chondrocytes from adipose tissue stem cells (autologous) and well attached to a bone phase (bone bovine decellularized) to mimic the structure of the nature of native tissue and to promote the cartilage regeneration in a model of joint damage in pigs. Biphasic scaffolds were assembled by fibroin crystallization with methanol. The histological and ultrastructural architectures were evaluated by optical and scanning electron microscopy respectively. Mechanical tests were conducted to evaluate Young's modulus of the implant. For the biological evaluation, pre-chondrocytes were loaded onto the scaffolds and cellular adhesion, proliferation, and gene expression analysis of cartilage extracellular matrix components was performed. The scaffolds that were cellularized and matured for 10 days were implanted into critical 3 mm in diameter and 9-mm in depth osteochondral defects in a porcine model (n=4). Three treatments were applied per knee: Group 1: monophasic cellular scaffold (MS) (single chondral phase), group 2: biphasic scaffold, cellularized only in the chondral phase (BS1), group 3: BS cellularized in both bone and chondral phases (BS2). Simultaneously, a control without treatment was evaluated. After 4 weeks of surgery, integration and regeneration tissues were analyzed by x-rays, histology and immunohistochemistry evaluation. The mechanical assessment showed that the acellular biphasic composites exhibited Young's modulus of 805.01 kPa similar to native cartilage (400-800 kPa). In vitro biological studies revealed the chondroinductive ability of the biphasic implant, evidenced by an increase in sulfated glycosaminoglycan (GAGs) and type II collagen, both secreted by the chondrocytes cultured on the scaffold during 28 days. No evidence of adverse or inflammatory reactions was observed in the in vivo trial; however, In group 1, the defects were not reconstructed. In group 2 and 3 a good integration of the implant with the surrounding tissue was observed. Defects in group 2 were fulfilled by hyaline cartilage and normal bone. Group 3 defects showed fibrous repair tissue. In conclusion; our findings demonstrated the efficacy of biphasic and bioactive scaffold based on silk fibroin, which entwined chondroinductive features and biomechanical capability with appropriate integration with the surrounding tissue, representing a promising alternative for osteochondral tissue-engineering applications.

Keywords: biphasic scaffold, extracellular cartilage matrix, silk fibroin, osteochondral tissue engineering

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Authors: Ntalamagka N., Sanchis-Benlloch P. J., Mayoral-Serrano R., Tena-Medialdea J., García-March J. R.

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The pen shell Pinna nobilis population has declined dramatically since 2016 due to die-off events observed in the whole extent of the Mediterranean Sea associated with the protozoan Haplosporidium pinnae. As of 2019, it is considered a critically endangered species. Due to its ecological importance and its endangered status, several initiatives have been developed for its salvation and recovery. This research is an effort to understand and control its reproduction under captivity. As a limited number of Pinna nobilis individuals could be used for experimentation, the possibility of using the Pinna rudis as a model animal was explored. The molecular mechanism that regulates the reproduction of both species is unknown; consequently, transcriptomic analysis was performed to identify neuropeptides that are expressed in the key regulatory tissues of the visceral ganglia and gonads of both species. Neuropeptides form an important group of signaling peptides that regulate reproductive, behavioral and physiological functions in molluscs. In total, 17 neuropeptide precursors were identified in P. nobilis and 14 in P. rudis transcriptomes; 14 of them were identical in both species. This affinity verified the genetic similarity of these species at the reproduction level. APGWamide, buccalin, ELH and GnRH were tested in P. rudis and demonstrated their capacity to advance gonadal maturation and trigger spawning while spawning was recorded in P. nobilis after the usage of APGWamide and buccalin. The neuropeptides were administered using intramuscular injection and cholesterol implants following relative literature as well as a new method was developed for external administration without the use of anesthesia using a mathematical model. The know-how of this research will not only lead to the survival of the species but also will narrow the horizons of broodstock conditioning of other similar species.

Keywords: neuropeptides, Pinna nobilis, reproduction, transcriptomics

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Authors: Po-Jung Huang, Chi-Ching Lee, Bertrand Chin-Ming Tan, Yuan-Ming Yeh, Julie Lichieh Chu, Tin-Wen Chen, Cheng-Yang Lee, Ruei-Chi Gan, Hsuan Liu, Petrus Tang

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Whole-exome sequencing focuses on the protein coding regions of disease/cancer associated genes based on a priori knowledge is the most cost-effective method to study the association between genetic alterations and disease. Recent advances in high throughput sequencing technologies and proteomic techniques has provided an opportunity to integrate genomics and proteomics, allowing readily detectable mutated peptides corresponding to mutated genes. Since sequence database search is the most widely used method for protein identification using Mass spectrometry (MS)-based proteomics technology, a mutant proteome database is required to better approximate the real protein pool to improve disease-associated mutated protein identification. Large-scale whole exome/genome sequencing studies were launched by National Cancer Institute (NCI), Broad Institute, and The Cancer Genome Atlas (TCGA), which provide not only a comprehensive report on the analysis of coding variants in diverse samples cell lines but a invaluable resource for extensive research community. No existing database is available for the collection of mutant protein sequences related to the identified variants in these studies. CMPD is designed to address this issue, serving as a bridge between genomic data and proteomic studies and focusing on protein sequence-altering variations originated from both germline and cancer-associated somatic variations.

Keywords: TCGA, cancer, mutant, proteome

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

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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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Authors: Hanaa H. Abd El Baky, Gamal S. El Baroty, Eman A. Ibrahem

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Microalgae Scenedesmus obliquus, the carotenoides (astaxanine and β-caroteine) were identified as the major bioactive constituents. In this work we prepared functional pre-biotic cakes to increase general mental health. Functional cakes were formulated by adding algal caroteinods at 2 and 4 mg/100g to flower and the cakes were storage for 20 days. Oxidative stability of both function cakes products were examined during storage periods by DPPH and TBA assays, and the results revealed that both values in function food products were significantly much low than that in untreated food products. Data of sensory evaluation revealed that treated biscuit and cakes with algae or algae extracts were significantly acceptable as control for main sensory characteristics (colour, odour/aroma, flavour, texture, the global appreciation, and overall acceptability). Thus, it could be concluded that functional biscuits and cakes (very popular and well balanced nutritional food) had good sensory and nutritional profiles and can be developed as new niche food market.

Keywords: Scenedesmus obliquus, carotenoids, functional cakes antioxidant, nutritional profiles

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Authors: Mahesh Pal, Tripti Mishra, Chandana Rao, Dalip Upreti

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Cancer has been considered to be a very dreadful disease. Holmskioldia sanguinea is a large climbing shrub found in the Himalayas at an altitude of 5,000 ft and preliminary investigation showed the excellent yield of andrographolide and subjected for the anticancer activity. Protective effect of Holmskioldia sanguinea leaf ethanolic extract has been investigated against Ehrlich ascites carcinoma (EAC) and Daltons ascites lymphoma (DAL) in Swiss albino mice to evaluate the possible mechanism of action. The enzymatic antioxidant status was studied on tumor bearing mice, which shows the potential of the compound to possess significant free radical scavenging property and revealed significant tumor regression and prolonged survival time. The isolated bioactive molecule andrographolide from Holmskioldia sanguinea yields (2.5%) in subject to HPTLC/HPLC analysis. The cellular defense system constituting the superoxide dismutase, catalyses was enhanced whereby the lipid peroxidation content was restricted to a larger extent. The Holmskioldia sanguinea is a new source of andrographolide and demonstrated the potency in treatment of cancer.

Keywords: Holmskioldia sanguinea, tumor, mice, andrographolide

Procedia PDF Downloads 234

Authors: Bangoura Issa, Ji-Young Kang, M. T. H. Chowdhury, Ji-Eun Lee, Yong-Ki Hong

Abstract:

Investigation was carried out for the production of value-added abalone Haliotis discus hannai containing bioactive phlorotannin by feeding phlorotannin-rich seaweed Eisenia bicyclis 2 weeks prior to harvesting. Accumulation of phlorotannins was proceded by feeding with E. bicyclis after 4 days of starvation. HPLC purification afforded two major phlorotannins. Mass spectrometry and 1H-nuclear magnetic resonance analysis clarified their structures to be as 7-phloroeckol and eckol. Throughout the feeding period of 20 days, 7-phloroeckolol was accumulated in the muscle (foot muscle tissue) up to 0.18±0.12 mg g-1 dry weight of tissue after 12 days. Eckol reached 0.21±0.03 mg g-1 dry weight of tissue after 18 days. By feeding Laminaria japonica as reference, abalone showed no detection of phlorotannins in the muscle tissue. Seaweed consumption and growth rate of abalone revealed almost similar when feed with E. bicyclis or L. japonicain 20 days. Phlorotannins reduction to half-maximal accumulation values took 1.0 day and 2.7 days for 7-phloroeckol and eckol respectively, after replacing the feed to L. japonica.

Keywords: abalone, accumulation, eisenia bicyclis, phlorotannins

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Authors: Alia Ragheb

Abstract:

Philodendron species were reported in traditional medicine for the treatment of several diseases. From the 70% methanol extract of the aerial parts of Philodendron bipinnatifidum Schott ex Endl, nine flavonoid compounds were isolated and identified for the first time; saponarin, genkwanin 8-C-(2′′-O-β-glucopyranosyl)-β-glucopyranoside, apigenin 6-C-(2′′-O-β-glucopyranosyl)-β-glucopyranoside, schaftoside, swertisin, swertiajaponin, isoswertisin, isorhamnetin 3-O-(2′′-acetyl)-β-glucopyranoside and apigenin. Characterization of the plant was achieved using chromatographic, physical, chemical, spectroscopic, and spectrometric techniques. The 70% methanol aerial parts extract and the methanol fraction of the plant were in vivo screened for their acute anti-inflammatory, antipyretic and analgesic effects where significant effects were exhibited compared to that of reference drugs. From the reported literature, these biological activities could be attributed to its phenolic constituent. The 70% methanol aerial parts and successive extracts, as well as some pure isolated flavonoid compounds, were in vitro investigated for their antioxidant, antimicrobial and cytotoxic activities.

Keywords: antioxidant, araceae, cytotoxicity, flavonoids

Procedia PDF Downloads 163

Authors: Shekhar U. Kadam, S. K. Pankaj, Brijesh K. Tiwari, P. J. Cullen, Colm P. O’Donnell

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Brown seaweed L. hyperborea is a rich source of phenolic compounds with antioxidant and antimicrobial properties. The aim of this work was to study the effect of incorporation of L. hyperborea extract to bovine gelatin film on the physicochemical and antioxidant properties of film. Films with fraction of 25% by weight of bovine gelatin sample were cast with addition of glycerol as a plasticizer. The total phenolic content and antioxidant activity of the films showed higher levels with addition of seaweed extract. Also film appearance properties such as film thickness, color and light transparency were evaluated. Film appearance was slightly modified whereas microstructure of films showed rough patches at 50% level of extract in the film. Hydrophilicity and glass transition temperature of the films also increased with increased level of seaweed extract. It was found that seaweed extract can be incorporated within gelatin and casein for development of biofunctional films.

Keywords: Laminaria hyperborea, ultrasound, seaweed extract, bovine gelatin film, antioxidant, phenolic compounds

Procedia PDF Downloads 500

Authors: Tarnveer Kaur

Abstract:

Amino acids, as fundamental structural units of peptides and proteins, have an important role in biological systems by affecting solubility, denaturation, and activity of biomolecules. A study of these effects on thermophysical properties of model compounds in the presence of electrolytes solutions provides information about solute-solvent and solute-solute interactions on biomolecules. Ionic liquids (ILs) as organic electrolytes and green solvents are composed of an organic cation and an inorganic anion, which are liquid at ambient conditions. In the past decade, extensive investigations showed that the use of ILs as reaction media for processes involving biologically relevant compounds is promising in view of their successful application in kinetic resolution, biocatalysis, biosynthesis, separation, and purification processes. The scope of this information is valuable to explore the interactions of amino acids in ILs. To reach this purpose, apparent molar volumes of glycine/L-alanine in aqueous solutions of 1-butyl-3-methylimidazolium bromide/chloride were determined from precise density measurements at temperatures T = (288.15-318.15) K and at atmospheric pressure. Positive values for all the studied amino acids indicate the dominance of hydrophilic-ionic interactions between amino acids and Ionic liquids. The effect of temperature on volumetric properties of glycine/L-alanine in solutions has been determined from the partial molar expansibility and second-order partial molar expansibility. Further, volumetric interaction parameters and hydration number have been calculated, which have been interpreted in terms of possible solute-solvent interactions.

Keywords: ILs, amino acids, volumetric properties, hydration numbers

Procedia PDF Downloads 149

Authors: Ghulam Hassan Abbasi, Moazzam Jamil, Ghazala Akhtar, M.Anwar-ul-Haq

Abstract:

Heavy metals are significant pollutants in environment and their toxicity is a problem for survival of living things while salicylic acid (SA) is signaling and ubiquitous bioactive molecule that regulates cellular mechanism in plants under stress condition. Therefore, exogenous application of salicylic acid (SA) under chromium stress in two chickpea varieties were investigated in hydroponic experiment with five treatments comprising of control, 5 µM Cr + 5 mM SA, 5µM Cr + 10 mM SA, 10µM Cr + 5 mM SA, and 10µM Cr + 10 mM SA. Results revealed that treatments of plants with 10 mM SA application under both 5 µM Cr and 10 µM Cr stress resulted in maximum improvement in plant morphological attributes (root and shoot length, root and shoot fresh and dry weight, membrane stability index and relative water contents) relative to 5 mM SA application in both chickpea varieties. Results regarding Cr concentration showed that Cr was more retained in roots followed by shoots and maximum reduction in Cr uptake was observed at 10 mM SA application. Chickpea variety BRC-61 showed maximum growth and least concentration of Cr in root and shoot relative to BRC-390 variety.

Keywords: chromium, Chickpea, salicylic acid, growth

Procedia PDF Downloads 489

Authors: Paiwan Buachan, Maneekarn Namsa-Aid, Suchada Jongrungruangchok, Foengchat Jarintanan, Wanlaya Uthaisang-Tanechpongtamb

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Lung cancer or pulmonary carcinoma is the uncontrolled growth of abnormal cells in one or both of the lungs. These abnormal cells can spread to other organs of the body through lymphatic system or bloodstream which is called metastatic stage that leading cause of cancer death. Terrein (C₈H₁₀O₃; MW= 154.06 kDa) is a secondary bioactive fungal metabolite, which was isolated from the Aspergillus terreus. In this study, we investigated the effects of terrein on the inhibition of human lung cancer cell proliferation and metastasis. The A549 human non-small cell lung cancer cell line was used as a model. Terrein significantly inhibited lung cancer cell proliferation measuring by a colorimetric MTT assay (IC₅₀ 0.32 mM) and significantly inhibited metastatic processes including migration, invasion, and adhesion that determined by wound healing assay, transwell assay, and adhesion assay, respectively. These findings indicate that terrein could be a potential therapeutic agent for lung cancer.

Keywords: terrein, lung cancer, anticancer, antimetastatic

Procedia PDF Downloads 147

Authors: Zahia Kabouche, Ouissem Gherboudj, Naima Boutaghane, Ahmed Kabouche, Laurence Voutquenne-Nazabadioko

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Chrysanthemum herbs (Asteraceae) are extensively used as food additives and in folk medicine. Anti-cancer, anti-human immunodeficiency virus type 1 (HIV-1), anti-inflammatory, antinociceptive and antiproliferative activities as well as antioxidant effects have been reported for Chrysanthemum species. We report the isolation and identification of flavonoids and new and known terpenoids from the endemic species, C. macrocarpum and C. deserticolum “guertoufa”, used in Algerian Sahara as tea drinks and in “couscous” and soups “Chorba”. Structures of the isolated compounds were established by 1-D and 2-D homo and hetero-nuclear NMR (1H, 13C, COSY, HSQC, HMBC, and NOESY), mass spectrometry, UV and comparison with literature data. C. deserticolum extracts were tested by four methods to identify the antioxidant activity namely, ABTS•+, DPPH• scavenging, CUPRAC and ferrous-ions chelating activity methods. Anti-inflammatory, antinociceptive, antiproliferative and antioxidant activities of C. macrocarpum extracts and isolated compounds are also reported here.

Keywords: Chrysanthemum macrocarpum, C. deserticolum, flavonoids, terpenoids, antioxidant, anti-inflammatory, anti-proliferative

Procedia PDF Downloads 316