Search results for: pea protein isolate

Authors: Eitedal Daoud, Reda M.Daoud, Khaled Abdel-Wahhab, Maha M.Saber, Lobna Saber

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Background: Cis-platin is a widely used chemotherapeutic drug to treat many malignant disorders including head and neck malignancies. Oto-nephrotxicity is an important and dose - limiting side effect of cis - platin therapy. Nowadays, more attention had been paid to oto-toxicity caused with cis-platin. Aim of the Work: This study was designed to investigate the potential protective effect of Whey protein (WP) against cis-platin induced ototoxicity compared to the effect of N-acetylcysteine (NAC) in rats. Methodology: Male albino rats were randomly divided into 6 groups: untreated rats (control), rats orally treated with whey protein (1g/kg b.w/day) for seven executive days, rats treated orally with N-acetylcysteine (500 mg/kgb.w /day) for seven executive days, rates intoxicated intraperitoneal (ip) with cis- platin (10 mg/kgb.w. once), rats treated with whey protein (1g/kgb.w./day) for seven executive days) followed by one injection (ip) of cis-platin(10 mg/kg b.w.) one hour after the last oral administration of whey protein, rats treated with N- acetylcysteine (for seven executive days followed by one injection (ip) of cis-platin (10 mg/kgb.w) one hour after the last oral administration of N-acetylcysteine. The organ of Corti, the stria vascularis and spiral ganglia were visualized by light microscopy at different magnifications. Results: Cis-platin intoxicated animals showed a significant decrease in serum level of total antioxidant capacity (TAC),with inhibition in the activity of serum glutathione-s transferase(GST) and paraoxonnase-1 (PON-1) in comparison with control. Group treated with either NAC or WP with cis-platin showed significant elevation in the activity of both GST & PON-1 with increased serum level of TAC when compared with cis-platin intoxicated rats. Animals treated with NAC or WP with cis-platin compared to those treated with cis-platin alone showed marked degree of improvement towards control rats as there was significant drop in the serum level of cortecosterone, nitric oxide (NO), and melandialdehyde (MDA).Histopathologic, in NAC pretreated group there was no changes in stria vascularis or spiral ganglia. In group pretreated with WP, there was no histopathologic alteration detected in the organ of Corti and Reissers membrane but oedema and haemorrhage were founded in the stria vascularis in small focal manner. Conclusion: Our finding showed that Whey protein is a natural dietary supplement product proved its ability of protection of anti-oxidant system and the cochlea against cis-platin induced ototoxicity.

Keywords: anti-oxidant, cis-platin, N-acetylcysteine, ototoxicity, whey protein

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Authors: Chieh-Chung Tsou, Chun-Min Lo, Yeh-Shiu Chu

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Cell’s mechanical forces provide important physical cues in regulation of proper cellular functions, such as cell differentiation, proliferation and migration. It is believed that adhesive forces generated by cell-cell interaction are able to transmit to the interior of cell through filamentous cortical cytoskeleton. Prominent among other membrane receptors, Cadherins are prototypical adhesive molecules able to generate remarkable forces to regulate intercellular adhesion. However, the mechanistic steps of mechano-transduction in Cadherin-mediated adhesion remain very controversial. We are interested in understanding how Cadherin protein complexes enable force generation and transmission at cell-cell contact in the initial stage of intercellular adhesion. For providing a better control of time, space, and substrate stiffness, in this study, a combination of protein micropattern, micropipette manipulation, and traction force microscopy is used. Pair micropattern with different forms confines cell spreading area and the gaps in pairs varied from 2 to 8 microns are applied for monitoring the forces that cell pairs generated, measured by traction force microscopy. Moreover, cell clones obtained from epithelial cells undergone genome editing are used to score the importance for known components of Cadherin complexes in force generation. We believe that our results from this combinatory mechanobiological method will provide deep insights on understanding the biophysical principle governing mechano- transduction of Cadherin-mediated intercellular adhesion.

Keywords: cadherin, intercellular adhesion, protein micropattern, traction force microscopy

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Authors: L. A. Bosnea, T. Moschakis, C. Biliaderis

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Two probiotic strains of Lactobacillus paracasei subsp. paracasei (E6) and Lactobacillus paraplantarum (B1), isolated from traditional Greek dairy products, were microencapsulated by complex coacervation using whey protein isolate (WPI, 3% w/v) and gum arabic (GA, 3% w/v) solutions mixed at different polymer ratio (1:1, 2:1 and 4:1). The effect of total biopolymer concentration on cell viability was assessed using WPI and GA solutions of 1, 3 and 6% w/v at a constant ratio of 2:1. Also, several parameters were examined for optimization of the microcapsule formation, such as inoculum concentration and the effect of ionic strength. The viability of the bacterial cells during heat treatment and under simulated gut conditions was also evaluated. Among the different WPI/GA weight ratios tested (1:1, 2:1, and 4:1), the highest survival rate was observed for the coacervate structures made with the ratio of 2:1. The protection efficiency at low pH values is influenced by both concentration and the ratio of the added biopolymers. Moreover, the inoculum concentration seems to affect the efficiency of microcapsules to entrap the bacterial cells since an optimum level was noted at less than 8 log cfu/ml. Generally, entrapment of lactobacilli in the complex coacervate structure enhanced the viability of the microorganisms when exposed to a low pH environment (pH 2.0). Both encapsulated strains retained high viability in simulated gastric juice (>73%), especially in comparison with non-encapsulated (free) cells (<19%). The encapsulated lactobacilli also exhibited enhanced viability after 10–30 min of heat treatment (65oC) as well as at different NaCl concentrations (pH 4.0). Overall, the results of this study suggest that complex coacervation with WPI/GA has a potential to deliver live probiotics in low pH food systems and fermented dairy products; the complexes can dissolve at pH 7.0 (gut environment), releasing the microbial cells.

Keywords: probiotic, complex coacervation, whey, encapsulation

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Authors: M. Latha, Achintya K. Dolui, P. Vijayaraj

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Vegetable oils mainly triacylglycerol (TAG) are an essential nutrient in the human diet as well as one of the major global commodity. There is a pressing need to enhance the yield of oil production to meet the world’s growing demand. Oil content is controlled by the balance between synthesis and breakdown in the cells. Several studies have established to increase the oil content by the overexpression of oil biosynthetic enzymes. Interestingly the significant oil accumulation was observed with impaired TAG hydrolysis. Unfortunately, the structural, as well as the biochemical properties of the lipase enzymes, is widely unknown, and so far, no candidate gene was identified in seeds except sugar-dependent1 (SDP1). Evidence has shown that SDP1directly responsible for initiation of oil breakdown in the seeds during germination. The present study is the identification of seed-specific acyl-hydrolases by activity based proteome profiling (ABPP) using Arabidopsis thaliana as a model system. The ABPP reveals that around 8 to 10 proteins having the serine hydrolase domain and are expressed during germination of Arabidopsis seed. The N-term sequencing, as well as LC-MS/MS analysis, was performed for the differentially expressed protein during germination. The coding region of the identified proteins was cloned, and lipases activity was assessed with purified recombinant protein. The enzyme assay was performed against various lipid substrates, and we have observed the acylhydrolase activity towards lysophosphatidylcholine and monoacylglycerol. Further, the functional characteristic of the identified protein will reveal the physiological significance the enzyme in oil accumulation.

Keywords: lipase, lipids, vegetable oil, triacylglycerol

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Authors: Jineetkumar Gawad, Chandrakant Bonde

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Tuberculosis (TB) is a major worldwide concern whose control has been exacerbated by HIV, the rise of multidrug-resistance (MDR-TB) and extensively drug resistance (XDR-TB) strains of Mycobacterium tuberculosis. The interest for newer and faster acting antitubercular drugs are more remarkable than any time. To search potent compounds is need and challenge for researchers. Here, we tried to design lead for inhibition of Decaprenyl phosphoryl-β-D-ribose 2′-epimerase (DprE1) enzyme. Arabinose is an essential constituent of mycobacterial cell wall. DprE1 is a flavoenzyme that converts decaprenylphosphoryl-D-ribose into decaprenylphosphoryl-2-keto-ribose, which is intermediate in biosynthetic pathway of arabinose. Latter, DprE2 converts keto-ribose into decaprenylphosphoryl-D-arabinose. We had a selection of 23 compounds from azaindole series for computational study, and they were drawn using marvisketch. Ligands were prepared using Maestro molecular modeling interface, Schrodinger, v10.5. Common pharmacophore hypotheses were developed by applying dataset thresholds to yield active and inactive set of compounds. There were 326 hypotheses were developed. On the basis of survival score, ADRRR (Survival Score: 5.453) was selected. Selected pharmacophore hypotheses were subjected to virtual screening results into 1000 hits. Hits were prepared and docked with protein 4KW5 (oxydoreductase inhibitor) was downloaded in .pdb format from RCSB Protein Data Bank. Protein was prepared using protein preparation wizard. Protein was preprocessed, the workspace was analyzed using force field OPLS 2005. Glide grid was generated by picking single atom in molecule. Prepared ligands were docked with prepared protein 4KW5 using Glide docking. After docking, on the basis of glide score top-five compounds were selected, (5223, 5812, 0661, 0662, and 2945) and the glide docking score (-8.928, -8.534, -8.412, -8.411, -8.351) respectively. There were interactions of ligand and protein, specifically HIS 132, LYS 418, TRY 230, ASN 385. Pi-pi stacking was observed in few compounds with basic Imidazo [4,5-b] pyridine ring. We had basic azaindole ring in parent compounds, but after glide docking, we received compounds with Imidazo [4,5-b] pyridine as a basic ring. That might be the new lead in the process of drug discovery.

Keywords: DprE1 inhibitors, in silico drug designing, imidazo [4, 5-b] pyridine, lead, tuberculosis

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Authors: M. R. Jokanović, V. M. Tomović, M. T. Jović, S. B. Škaljac, B. V. Šojić, P. M. Ikonić, T. A. Tasić

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Proximate (moisture, protein, total fat, total ash) and mineral (K, P, Na, Mg, Ca, Zn, Fe, Cu and Mn) composition of chicken giblets (heart, liver and gizzard) were investigated. Phosphorous content, as well as proximate composition, were determined according to recommended ISO methods. The content of all elements, except phosphorus, of the giblets tissues were determined using inductively coupled plasma-optical emission spectrometry (ICP-OES), after dry ashing mineralization. Regarding proximate composition heart was the highest in total fat content, and the lowest in protein content. Liver was the highest in protein and total ash content, while gizzard was the highest in moisture and the lowest in total fat content. Regarding mineral composition liver was the highest for K, P, Ca, Mg, Fe, Zn, Cu, and Mn, while heart was the highest for Na content. The contents of almost all investigated minerals in analysed giblets tissues of chickens from Vojvodina were similar to values reported in the literature, i.e. in national food composition databases of other countries.

Keywords: chicken giblets, proximate composition, mineral composition, inductively coupled plasma-optical emission spectrometry (ICP-OES)

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Authors: Hossein Rassi, Marjan Moradi Fard, Samaneh Niko

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Human papillomavirus type 16 and 18 are closely associated with the development of human cervical carcinoma, which is one of the most common causes of cancer death in women worldwide. At present, HPV type 18 accounts for about 34 % of all HPV infections in Iran and the most promising vaccine against HPV infection is based on the L1 major capsid protein. The L1 protein of HPV18 has the capacity to self-assemble into capsomers or virus-like particles (VLPs) that are non-infectious, highly immunogenic and allowing their use in vaccine production. The methylotrophic yeast Pichia pastoris is an efficient and inexpensive expression system used to produce high levels of heterologous proteins. In this study we expressed HPV18 L1 VLPs in P. pastoris. The gene encoding the major capsid protein L1 of the high-risk HPV type 18 was isolated from Iranian patient by PCR and inserted into pTG19-T vector to obtain the recombinant expression vector pTG19-HPV18-L1. Then, the pTG19-HPV18-L1 was transformed into E. coli strain DH5α and the recombinant protein HPV18 L1 was expressed under IPTG induction in soluble form. The HPV18 L1 gene was excised from recombinant plasmid with XhoI and EcoRI enzymes and ligated into the yeast expression vector pPICZα linearized with the same enzymes, and transformed into P. pastoris. Induction and expression of HPV18 L1 protein was demonstrated by BMGY/BMMY and RT PCR. The parameters for induced cultivation for strain in P. pastoris KM71 with HPV16L1 were investigated in shaking flask cultures. After induced cultivation BMMY (pH 7.0) medium supplemented with methanol to a final concentration of 1.0% every 24 h at 37 degrees C for 96 h, the recombinant produced 78.6 mg/L of L1 protein. This work offers the possibility for the production of prophylactic vaccine for cervical carcinoma by P. pastoris for HPV-18 L1 gene. The VLP-based HPV vaccines can prevent persistent HPV18 infections and cervical cancer in Iran. The HPV-18 L1 gene was expressed successfully in E.coli, which provides necessary basis for preparing HPV-18 L1 vaccine in human. Also, HPV type 6 L1 proteins expressed in Pichia pastoris will facilitate the HPV vaccine development and structure-function study.

Keywords: Pichia pastoris, L1 virus-like particles, human papillomavirus type 18, biotechnology

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Authors: Jesus Hernandez, Zead Elzoeiry, Md. S. Islam, Abel E. Navarro

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The adsorption of bovine serum albumin (BSA) and human hemoglobin (Hb) on naturally-occurring adsorbents was studied to evaluate the potential recovery of proteins from meat industry residues. Spent peppermint tea (PM), powdered purple corn cob (PC), natural clay (NC) and chemically-modified clay (MC) were investigated to elucidate the effects of pH, adsorbent dose, initial protein concentration, presence of salts and heavy metals. Equilibrium data were fitted according to isotherm models, reporting a maximum adsorption capacity at pH 8 of 318 and 344 mg BSA/g of PM and NC, respectively. Moreover, Hb displayed maximum adsorption capacity at pH 5 of 125 and 143 mg/g of PM and PC, respectively. Hofmeister salt effect was only observed for PM/Hb system. Salts tend to decrease protein adsorption, and the presence of Cu(II) ions had negligible impacts on the adsorption onto NC and PC. Desorption experiments confirmed that more than 85% of both proteins can be recovered with diluted acids and bases. SEM, EDX, and TGA analyses demonstrated that the adsorbents have favorable morphological and mechanical properties. The long-term goal of this study aims to recover soluble proteins from industrial wastewaters to produce animal food or any protein-based product.

Keywords: adsorption, albumin, clay, hemoglobin, spent peppermint leaf

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Authors: Sanchir Erdenebayar, Namuuntsetseg Oyunbaatar

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Objectives: Peritoneal dialysis treatment is one of the important forms of kidney replacement therapy, and it has recently developed instantly in Mongolia for the past five years. Currently, more than 120 patients undergo peritoneal dialysis nationwide. These patients lack nutritional education, which predisposes them to protein deficiency and further impairs their quality of life. However, there is no study which is conducted among those about their dietary in Mongolia. Therefore, integrated nutrition information and educating them about dietary patterns to follow are urgently needed for PD patients. Methods: A cross-sectional study was carried out on 45 patients aged between 18 and 60 years who were undergoing CAPD at the biggest Medvic dialysis center in Ulaanbaatar. The knowledge of nutrition and food intake is assessed by interview based on a validated questionnaire prepared from KDIGO guidelines, semi-FFQ and a 24-hour dietary recall method. In addition, a biochemical blood test that includes total protein, albumin, calcium, phosphorus, potassium, and hemoglobin is used for an assessment of the patient’s current nutritional status. Results: Knowledge of nutritional status for CAPD was great, with 21.4% of patients and 78.65% having poor nutrition knowledge. The rate of mild to moderate malnutrition was 48.8% among research participants. Serum albumin was 38.4 ± 4.7 g/L, and total protein was 67.3±7.5g/l. Patients met 62.5± 26.5% of their daily intake nutritional requirement for calories and 72±40% of their nutritional requirement for protein. All patients’ energy intake was significantly /1328±304kcal/ lower than the energy requirement (2124±378kcal). Only 14.2% met the recommended dietary protein intake recommended to them of greater than 1.2 g/kg. Conclusions: As was established before, nutritional education has a vital positive impact on the health and nutritional status of peritoneal dialysis patients. The results of this study show that nutritional education programs are not enough adequate in peritoneal dialysis patients. There is a crucial priority to establish nutritional educational programs and guidelines for PD patients in Mongolia.

Keywords: renal diet, peritoneal dialysis, nutrition education, CKD diet

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Authors: Nuri Başpınar, Abdullah Başoğlu, Özgür Özdemir, Çağlayan Özel, FundaTerzi, Özgür Yaman

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Current research is targeting new molecular mechanisms that underlie non-alcoholic fatty liver disease (NAFLD) and associated metabolic disorders like nonalcoholic steatohepatitis (NASH). Forty New Zealand White rabbits have been used and fed a high protein (HP) and energy diet based on grains and containing 11.76 MJ/kg. Boron added to 3 experimental groups’ drinking waters (30 mg boron/L) as boron compounds. Biochemical analysis including boron levels, and nuclear magnetic resonance (NMR) based metabolomics evaluation, and mRNA expression of peroxisome proliferator-activated receptor (PPAR) family were performed. LDL-cholesterol concentrations alone were decreased in all the experimental groups. Boron levels in serum and feces were increased. Content of acetate was in about 2x higher for anhydrous borax group, at least 3x higher for boric acid group. PPARα mRNA expression was significantly decreased in boric acid group. Anhydrous borax attenuated mRNA levels of PPARα, which was further suppressed by boric acid. Boron supplementation decreased the degenerative alterations in hepatocytes. Except borax group other boron groups did not have a pronounced change in tubular epithels of kidney. In conclusion, high protein and energy diet leads hepatocytes’ degenerative changes which can be prevented by boron supplementation. Boric acid seems to precede in this effectiveness.

Keywords: high protein and energy diet, boron, metabolomics, transcriptomic

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Authors: F. Bellali, M. Kharroubi, M. Loutfi, N.Bourhim

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In Morocco, fish processing industry is an important source income for a large amount of byproducts including skins, bones, heads, guts and scales. Those underutilized resources particularly scales contain a large amount of proteins and calcium. Scales from Sardina plichardus resulting from the transformation operation have the potential to be used as raw material for the collagen production. Taking into account this strong expectation of the regional fish industry, scales sardine upgrading is well justified. In addition, political and societal demands for sustainability and environment-friendly industrial production systems, coupled with the depletion of fish resources, drive this trend forward. Therefore, fish scale used as a potential source to isolate collagen has a wide large of applications in food, cosmetic and bio medical industry. The main aim of this study is to isolate and characterize the acid solubilize collagen from sardine fish scale, Sardina pilchardus. Experimental design methodology was adopted in collagen processing for extracting optimization. The first stage of this work is to investigate the optimization conditions of the sardine scale deproteinization on using response surface methodology (RSM). The second part focus on the demineralization with HCl solution or EDTA. Moreover, the last one is to establish the optimum condition for the isolation of collagen from fish scale by solvent extraction. The basic principle of RSM is to determinate model equations that describe interrelations between the independent variables and the dependent variables.

Keywords: Sardina pilchardus, scales, valorization, collagen extraction, response surface methodology

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Authors: Shikha Masand, Sudesh Kumar Yadav

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Certain protein kinases have been shown to be crucial for plant cell signaling pathways associated with plant immune responses. Here we identified a horsegram [Macrotyloma uniflorum (Lam.) Verdc.] malectin-like leucine rich receptor-like protein kinase (RLK) gene MuRLK. The functional MuRLK protein preferentially binds to mannose and N-acetyl glucosamine residues. MuRLK exists in the cytoplasm and also localizes to the plasma membrane of plant cells via its N-terminus. Over-expression of MuRLK in Arabidopsis enhances the basal resistance to infection with Pseudomonas syringae pv. tomato, Alternaria brassicicola and Hyaloperonospora arabidopsidis, are associated with elevated ROS bursts, MAPK activation, thus ultimately leading to hypersensitive cell death. Moreover, salicylic acid-dependent and jasmonic acid-dependent defense responses are also enhanced in the MuRLK-overexpressed plants that lead to HR-induced cell death. Together, these results suggest that MuRLK plays a key role in the regulation of plant cell death, early and late defense responses after the recognition of microbial pathogens.

Keywords: horsegram, Pseudomonas syringae pv. tomato, MuRLK, ROS burst, cell death, plant defense

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Authors: Maysam Mard-Soltani, Mohamad Javad Rasaee, Saeed Khalili, Abdol Karim Sheikhi, Mehdi Hedayati

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Production of specific antibody responses against hTSH is a cumbersome process due to the high identity between the hTSH and the other members of the glycoprotein hormone family (FSH, LH and HCG) and the high identity between the human hTSH and host animals for antibody production. Therefore, two polyclonal antibodies were purified against two recombinant proteins. Four possible ELISA tests were designed based on these antibodies. These ELISA tests were checked against hTSH and other glycoprotein hormones, and their sensitivity and specificity were assessed. Bioinformatics tools were used to analyze the immunological properties. After the immunogen region selection from hTSH protein, c terminal of B hTSH was selected and applied. Two recombinant genes, with these cut pieces (first: two repeats of C terminal of B hTSH, second: tetanous toxin+B hTSH C terminal), were designed and sub-cloned into the pET32a expression vector. Standard methods were used for protein expression, purification, and verification. Thereafter, immunizations of the white New Zealand rabbits were performed and the serums of them were used for antibody titration, purification and characterization. Then, four ELISA tests based on two antibodies were employed to assess the hTSH and other glycoprotein hormones. The results of these assessments were compared with standard amounts. The obtained results indicated that the desired antigens were successfully designed, sub-cloned, expressed, confirmed and used for in vivo immunization. The raised antibodies were capable of specific and sensitive hTSH detection, while the cross reactivity with the other members of the glycoprotein hormone family was minimum. Among the four designed tests, the test in which the antibody against first protein was used as capture antibody, and the antibody against second protein was used as detector antibody did not show any hook effect up to 50 miu/l. Both proteins have the ability to induce highly sensitive and specific antibody responses against the hTSH. One of the antibody combinations of these antibodies has the highest sensitivity and specificity in hTSH detection.

Keywords: hTSH, bioinformatics, protein expression, cross reactivity

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Authors: Farramae Francisco, Rhoda Mae Simora, Sharon Nunal

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Deproteination and demineralization efficiencies of shrimp waste using two Lactobacillus species treated with different carbohydrate sources for chitin production, its chemical conversion to chitosan and the quality of chitin and chitosan produced were determined. Using 5% glucose and 5% cassava starch as carbohydrate sources, pH slightly increased from the initial pH of 6.0 to 6.8 and 7.2, respectively after 24 h and maintained their pH at 6.7 to 7.3 throughout the treatment period. Demineralization (%) in 5 % glucose and 5 % cassava was highest during the first day of treatment which was 82% and 83%, respectively. Deproteination (%) was highest in 5% cassava starch on the 3rd day of treatment at 84.4%. The obtained chitin from 5% cassava and 5% glucose had a residual ash and protein below 1% and solubility of 59% and 44.3%, respectively. Chitosan produced from 5% cassava and 5% glucose had protein content below 0.05%; residual ash was 1.1% and 0.8%, respectively. Chitosan solubility and degree of deacetylation were 56% and 33% in 5% glucose and 48% and 29% in 5% cassava, respectively. The advantage this alternative technology offers over that of chemical extraction is large reduction in chemicals needed thus less effluent production and generation of a protein-rich liquor, although the demineralization process should be improved to achieve greater degree of deacetylation.

Keywords: alternative carbon source, bioprocessing, lactic acid bacteria, waste utilization

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Authors: Bahadir Batar, Elif Serdal, Berna Erdal, Hasan Ogul

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Micro-ribonucleic acids (miRNAs) are small short non-coding ribonucleic acid molecules about 22 nucleotides long. miRNAs play a key role in response to chemotherapeutic agents. WW domain-containing oxidoreductase (WWOX) gene encodes a tumor suppressor protein. Loss or reduction of Wwox protein is observed in many breast cancer cases. WWOX protein deficiency is increased in triple-negative breast cancer (TNBC). TNBC is a heterogeneous, highly aggressive, and difficult to treat tumor type. WWOX loss contributes to resistance to cisplatin therapy in patients with TNBC. Here, the aim of the study was to investigate the potential role of miRNAs in cisplatin therapy resistance of WWOX-deficient TNBC cells. This was a cell culture study. miRNA expression profiling was analyzed by LightCycler 480 system. miRNA Set Enrichment Analysis tool was used to integrate experimental data with literature-based biological knowledge to infer a new hypothesis. Increased miR-182 and decreased miR-214 were significantly correlated with cisplatin resistance in WWOX-deficient TNBC cells. miR-182 and miR-214 may involve in cisplatin resistance of WWOX-deficient TNBC cells by deregulating the DNA repair, apoptosis, or protein kinase B signaling pathways. These data highlight the mechanism by which WWOX regulates cisplatin resistance of TNBC and the potential use of WWOX as a predictor biomarker for cisplatin resistance.

Keywords: cisplatin, microRNA, triple-negative breast cancer, WWOX

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Authors: Jin Boo Jeong

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In this study, we elucidated anti-cancer activity and potential molecular mechanism of DKC against human colorectal cancer cells. DKC-E70 suppressed the proliferation of human colorectal cancer cell lines such as HCT116, SW480, LoVo and HT-29. Although DKC-E70 decreased cyclin D1 expression in protein and mRNA level, decreased level of cyclin D1 protein by DKC-E70 occurred at the earlier time than that of cyclin D1 mRNA, which indicates that DKC-E70-mediated downregulation of cyclin D1 protein may be a consequence of the induction of degradation and transcriptional inhibition of cyclin D1. In cyclin D1 degradation, we found that cyclin D1 downregulation by DKC-E70 was attenuated in presence of MG132. In addition, DKC-E70 phosphorylated threonine-286 (T286) of cyclin D1 and T286A abolished cyclin D1 downregulation by DKC-E70. We also observed that DKC-E70-mediated T286 phosphorylation and subsequent cyclin D1 degradation was blocked in presence of the inhibitors of ERK1/2, p38 or GSK3β. In cyclin D1 transcriptional inhibition, DKC-E70 inhibited the expression of β-catenin and TCF4, and β–catenin/TCF-dependent luciferase activity. Our results suggest that DKC-E70 may downregulate cyclin D1 as one of the potential anti-cancer targets through cyclin D1 degradation by T286 phosphorylation dependent on ERK1/2, p38 or GSK3β, and cyclin D1 transcriptional inhibition through Wnt signaling. From these findings, DKC-E70 has potential to be a candidate for the development of chemoprevention or therapeutic agents for human colorectal cancer. This research was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education (NRF-2016R1D1A3B03931713).

Keywords: anticancer, calyx of persimmon, cyclin D1, Diospyros kaki Thunb., human colorectal cancer

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Authors: Vitheejongjaroen P., Jaisin Y., Pachekrepapol U., Taweechotipatr M.

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Lactic acid bacteria (LAB) are the most common type of microorganisms that had been used as probiotics also known for many beneficial health effects. The antioxidant activity of LAB is associated with numerous health-protective effects. This research aimed to investigate the antioxidant activity of lactic acid bacteria isolated from Thai sour pork sausage for their application in fermented milk products. Antioxidant activity determined by DPPH (2,2-diphenyl-1-picrylhydrazyl) radical scavenging assay showed that the isolate FN33-7, as 1 of 8 isolated exhibited scavenging activity in intact cell 5-7%, and supernatant 13-16%, intracellular cell free extract 42-48% respectively. This isolate was identified using 16S ribosomal DNA sequence analysis as Lactobacillus plantarum. The effect of milk fermented with L. plantarum FN33-7 on microbial count, pH and syneresis was assessed during refrigerated storage period of 28 days. The strain showed increased viability, pH level decreased, while syneresis increased. These results are similar to dairy products fermented with commercial starter cultures. Additionally, microstructure analysis of fermented milk by fluorescent microscopy showed that curd structure appeared to be dense and less porous in this fermented milk than commercial yogurt. The results of this study indicated that L. plantarum FN33-7 was a good probiotic candidate to be used in cultured milk products to reduce the risk of diseases caused by oxidative stress.

Keywords: Lactobacillus plantarum, probiotics, free radical, antioxidant, oxidative stress, fermented milk products

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Authors: Charalampos N. Bompas, Eleni Poulou-Sidiropoulou, Martina Samiotaki, Alexios Vlamis-Gardikas

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Ιn all living organisms, antioxidant defenses are orchestrated by the thioredoxin (Trx) and glutaredoxin (Grx) systems. The Trx system of Escherichia coli (E. coli) is comprised of Trx1 and Trx2, both reduced by thioredoxin reductase (TrxR). The Grx system consists of four Grxs (Grx1, Grx2, Grx3, and Grx4), all reduced by glutathione (GSH) except for Grx4, which is reduced by TrxR. Under normal conditions, the GSH reductase of the Grx system keeps GSH at its reduced state. NADPH+ provides the electrons for all reductions in the Trx and Grx systems. Although the role of the E. coli Trx system is widely known, the function of the Grx system reflects the main property of Grx1, which is the reduction of ribonucleotide reductase Ia (RRIa). E. coli Grx3 (encoded by grxC) may also reduce RRIa in vitro but with slow kinetics. The molecule may account for up to 0.4% of total soluble protein and has been the subject of extensive structural studies. Its biological function, however, remains unknown. Herein, affinity chromatography with monothiol Grx3 serving as bait was used to detect the interactions of Grx3 with other proteins. Different types of interactions were identified (covalent, weak, and strong non-covalent) that suggested novel functions for Grx3. In silico approaches were employed to validate selected interactions. In addition, total protein extracts from the null mutant for grxC and the wild-type strain were compared. The overall findings suggest that Grx3 is involved in various metabolic processes, protein synthesis, and stress responses, expanding the recognized functions of Grx3 beyond the possible reduction of RRIa.

Keywords: escherichia coli, glutaredoxin 3, interactome, thiol-disulfide oxidoreductase

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Authors: Lydia Tan, Philippe Lemey, Lieselot Houspie, Marco Viveen, Darren Martin, Frank Coenjaerts

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Introduction: Human respiratory syncytial virus (hRSV) is the leading cause of severe respiratory tract infections in infants under the age of two. Genomic substitutions and related evolutionary dynamics of hRSV are of great influence on virus transmission behavior. The evolutionary patterns formed are due to a precarious interplay between the host immune response and RSV, thereby selecting the most viable and less immunogenic strains. Studying genomic profiles can teach us which genes and consequent proteins play an important role in RSV survival and transmission dynamics. Study design: In this study, genetic diversity and evolutionary rate analysis were conducted on 36 RSV subgroup B whole genome sequences and 37 subgroup A genome sequences. Clinical RSV isolates were obtained from nasopharyngeal aspirates and swabs of children between 2 weeks and 5 years old of age. These strains, collected during epidemic seasons from 2001 to 2011 in the Netherlands and Belgium by either conventional or 454-sequencing. Sequences were analyzed for genetic diversity, recombination events, synonymous/non-synonymous substitution ratios, epistasis, and translational consequences of mutations were mapped to known 3D protein structures. We used Bayesian statistical inference to estimate the rate of RSV genome evolution and the rate of variability across the genome. Results: The A and B profiles were described in detail and compared to each other. Overall, the majority of the whole RSV genome is highly conserved among all strains. The attachment protein G was the most variable protein and its gene had, similar to the non-coding regions in RSV, more elevated (two-fold) substitution rates than other genes. In addition, the G gene has been identified as the major target for diversifying selection. Overall, less gene and protein variability was found within RSV-B compared to RSV-A and most protein variation between the subgroups was found in the F, G, SH and M2-2 proteins. For the F protein mutations and correlated amino acid changes are largely located in the F2 ligand-binding domain. The small hydrophobic phosphoprotein and nucleoprotein are the most conserved proteins. The evolutionary rates were similar in both subgroups (A: 6.47E-04, B: 7.76E-04 substitution/site/yr), but estimates of the time to the most recent common ancestor were much lower for RSV-B (B: 19, A: 46.8 yrs), indicating that there is more turnover in this subgroup. Conclusion: This study provides a detailed description of whole RSV genome mutations, the effect on translation products and the first estimate of the RSV genome evolution tempo. The immunogenic G protein seems to require high substitution rates in order to select less immunogenic strains and other conserved proteins are most likely essential to preserve RSV viability. The resulting G gene variability makes its protein a less interesting target for RSV intervention methods. The more conserved RSV F protein with less antigenic epitope shedding is, therefore, more suitable for developing therapeutic strategies or vaccines.

Keywords: drug target selection, epidemiology, respiratory syncytial virus, RSV

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Authors: Tamar Khardziani, Violeta Berikashvili, Amrosi Chkuaseli, Vladimir Elisashvili

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The main objectives of this proposal are to develop low-cost, innovative, and competitive technologies for the production of mycelial biomass of medicinal mushrooms as a natural food supplement for poultry. To fulfill this task, industrial strains of Lentinus edodes, Ganoderma lucidum, and Pleurotus ostreatus were used in this study. The solid-state fermentation (SSF) of wheat grains, wheat bran, and soy flour was performed in flasks and bags. Among nine mushroom strains, P. ostreatus 2191 appeared to be the most productive in protein biomass accumulation in the SSF of wheat bran. All mushrooms produced exopolysaccharide with the highest yield of 5-8 mg/mL depending on fungal strain and growth substrate. Supplementation of medium with 1% glycerol and 2-4% peptone favored mushroom growth and protein accumulation. Among inorganic nitrogen sources, KNO₃ also provided high biomass and protein production. The SSF of all growth substrates was accompanied by the secretion of cellulase and xylanase activities. The highest CMCase activity (12-13 U/g) was revealed in the cultivation of P. ostreatus 2191 using wheat bran as a growth substrate and ammonium sulfate or yeast extract as a nitrogen source, whereas the highest xylanase activity was detected in the fermentation of soy flour supplemented with peptone. Acknowledgments: This work was supported by the Shota Rustaveli National Science Foundation of Georgia (Grant number STEM-22-2077).

Keywords: mushrooms, plant raw materials, fermentation, biomass protein, cellulase

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Authors: Nihan Nugay, Nur Cicek Kekec, Kalman Toth, Turgut Nugay, Joseph P. Kennedy

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In recent years, polyurethane structures are paving the way for elastomer usage in biology, human medicine, and biomedical application areas. Polyurethanes having a combination of high oxidative and hydrolytic stability and excellent mechanical properties are focused due to enhancing the usage of PUs especially for implantable medical device application such as cardiac-assist. Currently, unique polyurethanes consisting of polyisobutylenes as soft segments and conventional hard segments, named as PIB-based PUs, are developed with precise NCO/OH stoichiometry (∽1.05) for obtaining PIB-based PUs with enhanced properties (i.e., tensile stress increased from ∽11 to ∽26 MPa and elongation from ∽350 to ∽500%). Static and dynamic mechanical properties were optimized by examining stress-strain graphs, self-organization and crystallinity (XRD) traces, rheological (DMA, creep) profiles and thermal (TGA, DSC) responses. Annealing procedure was applied for PIB-based PUs. Annealed PIB-based PU shows ∽26 MPa tensile strength, ∽500% elongation, and ∽77 Microshore hardness with excellent hydrolytic and oxidative stability. The surface characters of them were examined with AFM and contact angle measurements. Annealed PIB-based PU exhibits the higher segregation of individual segments and surface hydrophobicity thus annealing significantly enhances hydrolytic and oxidative stability by shielding carbamate bonds by inert PIB chains. According to improved surface and microstructure characters, greater efforts are focused on analyzing protein adsorption and calcification profiles. In biomedical applications especially for cardiological implantations, protein adsorption inclination on polymeric heart valves is undesirable hence protein adsorption from blood serum is followed by platelet adhesion and subsequent thrombus formation. The protein adsorption character of PIB-based PU examines by applying Bradford assay in fibrinogen and bovine serum albumin solutions. Like protein adsorption, calcium deposition on heart valves is very harmful because vascular calcification has been proposed activation of osteogenic mechanism in the vascular wall, loss of inhibitory factors, enhance bone turnover and irregularities in mineral metabolism. The calcium deposition on films are characterized by incubating samples in simulated body fluid solution and examining SEM images and XPS profiles. PIB-based PUs are significantly more resistant to hydrolytic-oxidative degradation, protein adsorption and calcium deposition than ElastEonTM E2A, a commercially available PDMS-based PU, widely used for biomedical applications.

Keywords: biomedical application, calcification, polyisobutylene, polyurethane, protein adsorption

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Authors: K. B. M. Islam, Syeeda Shiraj-Um-Mahmuda, Afroj Jahan, A. A. Bhuiyan

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In Bangladesh, the use of imported probiotics in poultry is gradually being increased. But surprisingly, no probiotic bacteria have been isolated yet in Bangladesh despite the existence of scavenging native poultry as potential source that is seemingly more resistant to GIT infection as well as other diseases. Therefore, the study was undertaken to isolate, identify and characterize the potential probiotic Lactobacillus and Bifidobacteria strains from Bangladeshi indigenous poultry, and to evaluate their suitability to use in poultry industry. Crop and cecal samples from 61 healthy indigenous birds were used to isolate potential probiotics strains following conventional cultural methods. A total of 216 isolates were identified following physical, biochemical and molecular methods that belonged to the genus Lactobacillus and Bifidobacteria. An auto-aggregation test was performed for 180 and 136 isolated lactobacilli and bifidobacteria strains, respectively. Twelve lactobacilli isolates and 7 bifidobacteria isolates were selected because of their convenient aggregation. In vitro tests including antibacterial activity, resistance to low pH, hemolytic activities etc. were performed for evaluation of probiotic potential of each strain. Under the in vitro conditions and with respects to the probiotic traits, three lactobacilli; LS16, LS45, LS133 and two bifidobacteria, BS21 and BS90 were found to be potential probiotic strains. Thus, they are proposed to be evaluated for their in vivo probiotic properties. If the proposed strains are found suitable as the probiotics to be used in commercial poultry industry, it is expected that the local probiotics would be more beneficial and would save the huge amount of money that Bangladesh spends every year for the importation of such materials from abroad.

Keywords: Bangladeshi poultry, gut microbiota, lactic acid bacteria, scavenging chicken, GIT health

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Authors: Md. Samsuddin Ansari, Ashish Arora

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Every year 10 million people fall ill with one of the oldest diseases known as tuberculosis, caused by Mycobacterium tuberculosis. The success of M. tuberculosis as a pathogen is because of its ability to persist in host tissues. Multidrug resistance (MDR) mycobacteria cases increase every day, which is associated with efflux pumps controlled at the level of transcription. The transcription regulators of MDR transporters in bacteria belong to one of the following four regulatory protein families: AraC, MarR, MerR, and TetR. Phenolic acid decarboxylase repressor (PadR), like a family of transcription regulators, is closely related to the MarR family. Phenolic acid decarboxylase repressor (PadR) was first identified as a transcription factor involved in the regulation of phenolic acid stress response in various microorganisms (including Mycobacterium tuberculosis H37Rv). Recently research has shown that the PadR family transcription factors are global, multifunction transcription regulators. Rv0047c is a PadR subfamily-1 protein. We are exploring the biophysical and structural characterization of Rv0047c. The Rv0047 gene was amplified by PCR using the primers containing EcoRI and HindIII restriction enzyme sites cloned in pET-NH6 vector and overexpressed in DH5α and BL21 (λDE3) cells of E. coli following purification with Ni2+-NTA column and size exclusion chromatography. We did DSC to know the thermal stability; the Tm (transition temperature) of protein is 55.29ºC, and ΔH (enthalpy change) of 6.92 kcal/mol. Circular dichroism to know the secondary structure and conformation and fluorescence spectroscopy for tertiary structure study of protein. To understand the effect of pH on the structure, function, and stability of Rv0047c we employed spectroscopy techniques such as circular dichroism, fluorescence, and absorbance measurements in a wide range of pH (from pH-2.0 to pH-12). At low and high pH, it shows drastic changes in the secondary and tertiary structure of the protein. EMSA studies showed the specific binding of Rv0047c with its own 30-bp promoter region. To determine the effect of complex formation on the secondary structure of Rv0047c, we examined the CD spectra of the complex of Rv0047c with promoter DNA of rv0047. The functional role of Rv0047c was characterized by over-expressing the Rv0047c gene under the control of hsp60 promoter in Mycobacterium tuberculosis H37Rv. We have predicted the three-dimensional structure of Rv0047c using the Swiss Model and Modeller, with validity checked by the Ramachandra plot. We did molecular docking of Rv0047c with dnaA, through PatchDock following refinement through FireDock. Through this, it is possible to easily identify the binding hot-stop of the receptor molecule with that of the ligand, the nature of the interface itself, and the conformational change undergone by the protein pattern. We are using X-crystallography to unravel the structure of Rv0047c. Overall the studies show that Rv0047c may have transcription regulation along with providing an insight into the activity of Rv0047c in the pH range of subcellular environment and helps to understand the protein-protein interaction, a novel target to kill dormant bacteria and potential strategy for tuberculosis control.

Keywords: mycobacterium tuberculosis, phenolic acid decarboxylase repressor, Rv0047c, Circular dichroism, fluorescence spectroscopy, docking, protein-protein interaction

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Authors: Kyung-Tae Lee, Li Li Wang, Kwang-Woo Jung, Yong-Sun Bahn

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Microtubules are involved in mechanical support, cytoplasmic organization as well as in a number of cellular processes by interacting with diverse microtubule-associated proteins (MAPs), such as plus-end tracking proteins, motor proteins, and tubulin-folding cofactors. A common feature of these proteins is the presence of a cytoskeleton-associated protein-glycine-rich (CAP-Gly) domain, which is evolutionarily conserved and generally considered to bind to α-tubulin to regulate functions of microtubules. However, there has been a dearth of research on CAP-Gly proteins in fungal pathogens, including Cryptococcus neoformans, which causes fatal meningoencephalitis globally. In this study, we identified five CAP-Gly proteins encoding genes in C. neoformans. Among these, Cgp1, encoded by CNAG_06352, has a unique domain structure that has not been reported before in other eukaryotes. Supporting the role of Cpg1 in microtubule-related functions, we demonstrate that deletion or overexpression of CGP1 alters cellular susceptibility to thiabendazole, a microtubule destabilizer, and Cgp1 is co-localized with cytoplasmic microtubules. Related to the cellular functions of microtubules, Cgp1 also governs maintenance of membrane stability and genotoxic stress responses. Furthermore, we demonstrate that Cgp1 uniquely regulates sexual differentiation of C. neoformans with distinct roles in the early and late stage of mating. Our domain analysis reveals that the CAP-Gly domain plays major roles in all the functions of Cgp1. Finally, the cgp1Δ mutant is attenuated in virulence. In conclusion, this novel CAP-Gly protein, Cgp1, has pleotropic roles in regulating growth, stress responses, differentiation and pathogenicity of C. neoformans.

Keywords: human fungal pathogen, CAP-Glycine protein, microtubule, meningoencephalitis

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Authors: Carolina V. Viegas, Monique Gonçalves, Gisel Chenard Diaz, Yordanka Reyes Cruz, Donato Alexandre Gomes Aranda

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To develop the massive cultivation of microalgae, it is necessary to isolate and characterize the species, improving genetic tools in search of specific characteristics. Therefore, the detection, identification and quantification of the compounds that compose the Scenedesmus sp. were prerequisites to verify the potential of these microalgae. The main objective of this work was to carry out the characterization of Scenedesmus sp. as to the content of ash, carbohydrates, proteins and lipids as well as the determination of the composition of their lipid classes and main fatty acids. The biomass of Scenedesmus sp, showed 15,29 ± 0,23 % of ash and CaO (36,17 %) was the main component of this fraction, The total protein and carbohydrate content of the biomass was 40,74 ± 1,01 % and 23,37 ± 0,95 %, respectively, proving to be a potential source of proteins as well as carbohydrates for the production of ethanol via fermentation, The lipid contents extracted via Bligh & Dyer and in situ saponification were 8,18 ± 0,13 % and 4,11 ± 0,11 %, respectively. In the lipid extracts obtained via Bligh & Dyer, approximately 50 % of the composition of this fraction consists of fatty compounds, while the other half is composed of an unsaponifiable fraction composed mainly of chlorophylls, phytosterols and carotenes. From the lowest yield, it was possible to obtain a selectivity of 92,14 % for fatty components (fatty acids and fatty esters) confirmed through the infrared spectroscopy technique. The presence of polyunsaturated acids (~45 %) in the lipid extracts indicated the potential of this fraction as a source of nutraceuticals. The results indicate that the biomass of Scenedesmus sp, can become a promising potential source for obtaining polyunsaturated fatty acids, carotenoids and proteins as well as the simultaneous obtainment of different compounds of high commercial value.

Keywords: microalgae, Desmodesmus, lipid classes, fatty acid profile, proteins, carbohydrates

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Authors: Preeyarach Whisetkhan, Malai Taweechotipatr, Ulisa Pachekrepapol

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Elevated level of blood cholesterol or hypercholesterolemia may lead to atherosclerosis and poses a major risk for cardiovascular diseases. Probiotics play a crucial role in human health, and probiotic bacteria that possesses bile salt hydrolase (BSH) activity can be used to lower cholesterol level of the host. The aim of this study was to investigate whether lactic acid bacteria (LAB) isolated from traditional Thai fermented foods were able to exhibit bile salt hydrolase activity and their use in fermented milk. A total of 28 isolates were tested for BSH activity by plate method on MRS agar supplemented with 0.5% sodium salt of taurodeoxycholic acid and incubated at 37°C for 48 h under anaerobic condition. The results showed that FN1-1 and FN23-3 isolates possessed strong BSH activity. FN1-1 and FN23-3 isolates were then identified for phenotype, biochemical characteristics, and genotype (16S rRNA sequencing). FN1-1 isolate showed 99.92% similarity to Lactobacillus pentosus DSM 20314(T), while FN23-3 isolate showed 99.94% similarity to Enterococcus faecium CGMCC1.2136 (T). Lactobacillus pentosus FN1-1 and Enterococcus faecium FN23-3 were tolerant of pH 3-4 and 0.3 and 0.8% bile. Bacterial count and pH of milk fermented with Lactobacillus pentosus FN1-1 at 37°C and 43°C were investigated. The results revealed that Lactobacillus pentosus FN1-1 was able to grow in milk, which led to decrease in pH level. Fermentation at 37°C resulted in faster growth rate than at 43 °C. Lactobacillus pentosus FN1-1 was a candidate probiotic to be used in fermented milk products to reduce the risk of high-cholesterol diseases.

Keywords: probiotics, lactic acid bacteria, bile salt hydrolase, cholesterol

Procedia PDF Downloads 132

Authors: Iftikhar A. Tayubi, Tabrej Khan, Mansoor M. Alsubei, Fahad A. Alsaferi

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LIZTOXD provides a single source of high-quality information about proteinaceous lizard toxins that will be an invaluable resource for pharmacologists, neuroscientists, toxicologists, medicinal chemists, ion channel scientists, clinicians, and structural biologists. We will provide an intuitive, well-organized and user-friendly web interface that allows users to explore the detail information of Lizard and toxin proteins. It includes common name, scientific name, entry id, entry name, protein name and length of the protein sequence. The utility of this database is that it can provide a user-friendly interface for users to retrieve the information about Lizard, toxin and toxin protein of different Lizard species. These interfaces created in this database will satisfy the demands of the scientific community by providing in-depth knowledge about Lizard and its toxin. In the next phase of our project we will adopt methodology and by using A MySQL and Hypertext Preprocessor (PHP) which and for designing Smart Draw. A database is a wonderful piece of equipment for storing large quantities of data efficiently. The users can thus navigate from one section to another, depending on the field of interest of the user. This database contains a wealth of information on species, toxins, toxins, clinical data etc. LIZTOXD resource that provides comprehensive information about protein toxins from lizard toxins. The combination of specific classification schemes and a rich user interface allows researchers to easily locate and view information on the sequence, structure, and biological activity of these toxins. This manually curated database will be a valuable resource for both basic researchers as well as those interested in potential pharmaceutical and agricultural applications of lizard toxins.

Keywords: LIZTOXD, MySQL, PHP, smart draw

Procedia PDF Downloads 139

Authors: Ngonidzashe Mangoma, Caroline Marigold Sebata

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The release of cyanide-rich effluents from gold mines, and other industries, into the environment, is a global concern considering the well-known metabolic effects of cyanide in all forms of life. Such effluents need to be treated to remove cyanide, among other pollutants, before their disposal. This study aimed at investigating the possible use of bacteria in the biological removal of cyanide from cyanide-rich effluents. Firstly, cyanide-degrading bacteria were isolated from gold mine effluents and characterised. The isolates were then tested for their ability to grow in the presence of cyanide and their tolerance to increasing levels of the compound. To evaluate each isolate’s cyanide-degrading activities, isolates were grown in the simulated and actual effluent, and a titrimetric method was used to quantify residual cyanide over a number of days. Cyanide degradation efficiency (DE) was then calculated for each isolate. Identification of positive isolates involved 16S rRNA gene amplification and sequence analysis through BLAST. Six cyanide-utilising bacterial strains were isolated. Two of the isolates were identified as Klebsiella spp. while the other two were shown to be different strains of Clostridium bifermentans. All isolates showed normal growth in the presence of cyanide, with growth being inhibited at 700 mg/L cyanide and beyond. Cyanide degradation efficiency for all isolates in the simulated effluent ranged from 79% to 97%. All isolates were able to remove cyanide from actual gold mine effluent with very high DE values (90 – 94%) being recorded. Isolates obtained in this study were able to efficiently remove cyanide from both simulated and actual effluent. This observation clearly demonstrates the feasibility of the biological removal of cyanide from cyanide-rich gold mine effluents and should, therefore, motivate research towards the possible large-scale application of this technology.

Keywords: cyanide effluent, bioremediation, Clostridium bifermentans, Klebsiella spp, environment

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Authors: Prasanna Ramachandran, Kareem Graham, Helena Kiefel, Sunit Jain, Todd DeSantis

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Microbes that reside inside the mammalian GI tract, commonly referred to as the gut microbiome, have been shown to have therapeutic effects in animal models of disease. We hypothesize that specific proteins produced by these microbes are responsible for this activity and may be used directly as therapeutics. To speed up the discovery of these key proteins from the big-data metagenomics, we have applied machine learning techniques. Using amino acid sequences of known epitopes and their corresponding binding partners, protein interaction descriptors (PID) were calculated, making a positive interaction set. A negative interaction dataset was calculated using sequences of proteins known not to interact with these same binding partners. Using Random Forest and positive and negative PID, a machine learning model was trained and used to predict interacting versus non-interacting proteins. Furthermore, the continuous variable, cosine similarity in the interaction descriptors was used to rank bacterial therapeutic candidates. Laboratory binding assays were conducted to test the candidates for their potential as therapeutics. Results from binding assays reveal the accuracy of the machine learning prediction and are subsequently used to further improve the model.

Keywords: protein-interactions, machine-learning, metagenomics, microbiome

Procedia PDF Downloads 345

Authors: Sharon N. Nuñal, May Flor S. Muegue, Nizzy Hope N. Cartago, Raymund B. Parcon, Sheina B. Logronio

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The antioxidant and anti-inflammatory potentials of Perna Viridis, Modiolus philippinarum, and Mytella charruana found in the Philippines were assessed. Mussel protein samples were hydrolyzed using trypsin, maturase, alcalase and pepsin at 1% and 2% concentrations and then fractionated through membrane filtration (<10 kDa and <30 kDa). Antioxidant assays showed that pepsin hydrolysate at 2% enzyme concentration exhibited the maximum activities for both 2,2-Diphenyl-1-picrylhydrazyl (DPPH) Radical Scavenging Activity (155-176 µM TE/mg protein) and 2,2-azinobis-(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) radical scavenging (67-68 µM TE/mg protein) assays while trypsin hydrolysate dominated the Ferric Reducing Antioxidant Power (FRAP) for the three mussel species. Lower molecular weight peptide fractions at <10 kDa exhibited better antioxidant activities than the higher molecular weight fractions. The anti-inflammatory activities of M. philippinarum and M. charruana showed comparable protein denaturation inhibition potentials with the highest in P. Viridis samples (98.93%). The 5-Lipoxygenase (5-LOX) inhibitory activities of mussel samples showed no significant difference with inhibition exceeding 70%. P. Viridis demonstrated the highest inhibition against Cyclooxygenase-2 (COX-2) at 56.19%, while the rest showed comparable activities. This study showed that the three mussel species are potential sources of bioactive peptides and lipids with antioxidant and anti-inflammatory properties.

Keywords: anti-inflammatory, antioxidant, bioactive properties, mussel

Procedia PDF Downloads 189