Search results for: protein profile
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 4328

Search results for: protein profile

3638 A Radioprotective Effect of Nanoceria (CNPs), Magnetic Flower-Like Iron Oxide Microparticles (FIOMPs), and Vitamins C and E on Irradiated BSA Protein

Authors: Hajar Zarei, AliAkbar Zarenejadatashgah, Vuk Uskoković, Hiroshi Watabe

Abstract:

The reactive oxygen species (ROS) generated by radiation in nuclear diagnostic imaging and radiotherapy could damage the structure of the proteins in noncancerous cells surrounding the tumor. The critical factor in many age-related diseases, such as Alzheimer, Parkinson, or Huntington diseases, is the oxidation of proteins by the ROS as molecular triggers of the given pathologies. Our studies by spectroscopic experiments showed doses close to therapeutic ones (1 to 5 Gy) could lead to changes of secondary and tertiary structures in BSA protein macromolecule as a protein model as well as the aggregation of polypeptide chain but without the fragmentation. For this reason, we investigated the radioprotective effects of natural (vitamin C and E) and synthetic materials (CNPs and FIOMPs) on the structural changes in BSA protein induced by gamma irradiation at a therapeutic dose (3Gy). In the presence of both vitamins and synthetic materials, the spectroscopic studies revealed that irradiated BSA was protected from the structural changes caused by ROS, according to in vitro research. The radioprotective property of CNPs and FIOMPs arises from enzyme mimetic activities (catalase, superoxide dismutase, and peroxidase) and their antioxidant capability against hydroxyl radicals. In the case of FIOMPs, a porous structure also leads to increased ROS recombination with each other in the same radiolytic track and subsequently decreased encounters with BSA. The hydrophilicity of vitamin C resulted in the major scavenging of ROS in the solvent, whereas hydrophobic vitamin E localized on the nonpolar patches of the BSA surface, where it did not only neutralize them thanks to the moderate BSA binding constant but also formed a barrier for diffusing ROS. To the best of our knowledge, there has been a persistent lack of studies investigating the radioactive effect of mentioned materials on proteins. Therefore, the results of our studies can open a new widow for application of these common dietary ingredients and new synthetic NPs in improving the safety of radiotherapy.

Keywords: reactive oxygen species, spectroscopy, bovine serum albumin, gamma radiation, radioprotection

Procedia PDF Downloads 86
3637 Effect of Inhibitor of the Angiotensin Converting Enzyme in the Mediterranean Flour Moth: Structural Parametrs of Cuticule and Ecdysteroid Amounts

Authors: S. Yezli-Touiker, L. Kirane-Amrani, N. Soltani-Mazouni

Abstract:

Ephestia kuehniella Zeller Lepidoptera, Pyralidae commonly called Mediterranean flour moth, is serious cosmopolitan pest of stored grain products, particularly flour Month. This species is also a source of allergen that causes asthma and rhinitis. Captopril is an inhibitor of angiotensin converting enzyme (ACE) it was tested in vivo by topical application on development of E. kuehniella. The compound is diluted in acetone and applied topically to newly emerged pupae (10mg/2ml). Report chitin protein of cuticule and ecdysteroid Amounts were determined in vivo. Results show that the captopril does not affect chitin protein of cuticule but traitment with captopril increase the hormonal production, the quantitative analysis reveals the presence of two peaks one at third and another at fifth day.

Keywords: Ephestia kuehniella, cuticule, hormone, captopril

Procedia PDF Downloads 356
3636 Nematicidal Activity of the Cell Extract from Penicillium Sp EU0013 and Its Metabolite Profile Using High Performance Liquid Chromatograpy

Authors: Zafar Iqbal, Sana Irshad Khan

Abstract:

Organic extract from newly isolated plant growth promoting fungus (PGPF) Penicillium sp EU0013 was subjected to bioassays including anti fungal (disc diffusion) cytotoxicity (brine shrimp lethality), herbicidal (Lemna minor) and nematicidal activities. Metabolite profile of the extract was also assessed using HPLC analysis with the aim to identify bioactive natural products in the extract as new drug candidate(s). The extract showed anti fungal potential against tested fungal pathogens. Growth of the Wilt pathogen Fusarium oxyosproum was inhibited up to 63% when compared to negative reference. Activity against brine shrimps was weak and mortality up to 10% was observed at concentration of 200 µg. mL-1. The extract exhibited no toxicity against Lemna minor frond at 200 µg. mL-1. Nematicidal activity was observed very potent against root knot nematode and LC50 value was calculated as 52.5 ug. mL-1 using probit analysis. Methodically assessment of metabolites profile by HPLC showed the presence of kojic acid (Rt 1.4 min) and aflatoxin B1 (Rt 5.9 min) in the mycellial extract as compared with standards. The major unidentified metabolite was eluted at Rt 8.6 along with other minor peaks. The observed high toxicity against root knot nematode was attributed to the unidentified compounds that make fungal extract worthy of further exploration for isolation and structural characterization studies for development of future commercial nematicidal compound(s).

Keywords: penicillium, nematicidal activity, metabolites, HPLC

Procedia PDF Downloads 446
3635 Use of Zikani’s Ribosome Modulating Agents for Treating Recessive Dystrophic & Junctional Epidermolysis Bullosa with Nonsense Mutations

Authors: Mei Chen, Yingping Hou, Michelle Hao, Soheil Aghamohammadzadeh, Esteban Terzo, Roger Clark, Vijay Modur

Abstract:

Background: Recessive Dystrophic Epidermolysis Bullosa (RDEB) is a genetic skin condition characterized by skin tearing and unremitting blistering upon minimal trauma. Repeated blistering, fibrosis, and scarring lead to aggressive squamous cell carcinoma later in life. RDEB is caused by mutations in the COL7A1 gene encoding collagen type VII (C7), the major component of anchoring fibrils mediating epidermis-dermis adherence. Nonsense mutations in the COL7A1 gene of a subset of RDEB patients leads to premature termination codons (PTC). Similarly, most Junctional Epidermolysis Bullosa (JEB) cases are caused by nonsense mutations in the LAMB3 gene encoding the β3 subunit of laminin 332. Currently, there is an unmet need for the treatment of RDEB and JEB. Zikani Therapeutics has discovered an array of macrocyclic compounds with ring structures similar to macrolide antibiotics that can facilitate readthrough activity of nonsense mutations in the COL7A1 and LAMB3 genes by acting as Ribosome Modulating Agents (RMAs). The medicinal chemistry synthetic advancements of these macrocyclic compounds have allowed targeting the human ribosome while preserving the structural elements responsible for the safety and pharmacokinetic profile of clinically used macrolide antibiotics. Methods: C7 expression was used as a measure of readthrough activity by immunoblot assays in two primary human fibroblasts from RDEB patients (R578X/R578X and R163X/R1683X-COL7A1). Similarly, immunoblot assays in C325X/c.629-12T > A-LAMB3 keratinocytes were used to measure readthrough activity for JEB. The relative readthrough activity of each compound was measured relative to Gentamicin. An imaging-based fibroblast migration assay was used as an assessment of C7 functionality in RDEB-fibroblasts over 16-20 hrs. The incubation period for the above experiments was 48 hrs for RDEB fibroblasts and 72 hours for JEB keratinocytes. Results: 9 RMAs demonstrated increased protein expression in both patient RDEB fibroblasts. The highest readthrough activity at tested concentrations without cytotoxicities increased protein expression up to 179% of Gentamicin (400 µg/ml), with favored readthrough activity in R163X/R1683X-COL7A1 fibroblasts. Concurrent with protein expression, fibroblast hypermotility phenotype observed in RDEB was rescued by reducing motility by ~35% to WT levels (the same level as 690 µM Gentamicin treated cells). Laminin β3 expression was also shown to be increased by 6 RMAs in keratinocytes to 33-83% of (400 µg/ml) Gentamicin. Conclusions: To date, 9 RMAs have been identified that enhance the expression of functional C7 in a mutation-dependent manner in two different RDEB patient fibroblast backgrounds (R578X/R578X and R163X/R1683X-COL7A1). A further 6 RMAs have been identified that enhance the readthrough of C325X-LAMB3 in JEB patient keratinocytes. Based on the clinical trial conducted by us with topical gentamycin in 2017, Zikani’s RMAs achieve clinically significant levels of read-through for the treatment of recessive dystrophic and Junctional Epidermolysis Bullosa.

Keywords: epidermolysis bullosa, nonsense mutation, readthrough, ribosome modulation

Procedia PDF Downloads 98
3634 Investigation of Nutritional Values, Sensorial, Flesh Productivity of Parapenaus longirostris between Populations in the Sea of Marmara and in the Northern Aegean Sea

Authors: Onur Gönülal, Zafer Ceylan, Gülgün F. Unal Sengor

Abstract:

The differences of Parapenaus longirostris caught from The North Aegean Sea and the Marmara Sea on proximate composition, sensorial analysis (for raw and cooked samples), flesh productivity of the samples were investigated. The moisture, protein, lipid, ash, carbohydrate, energy contents of shrimp caught from The North Aegean Sea were 74.92 ± 0.1, 20.32 ± 0.16, 2.55 ± 0.1, 2.13 ± 0.08, 0.08, 110.1 kcal/100g, respectively. The moisture, protein, lipid, ash, carbohydrate, energy contents of shrimp caught from Marmara Sea were 76.9 ± 0.02, 19.06 ± 0.03, 2.22 ± 0.08, 1.51 ± 0.04, 0.33, 102.77 kcal/100g, respectively. The protein, lipid, ash and energy values of the Northern Aegean Sea shrimp were higher than The Marmara Sea shrimp. On the other hand, The moisture, carbohydrate values of the Northern Aegean Sea shrimp were lower than the other one. Sensorial analysis was done for raw and cooked samples. Among all properties for raw samples, flesh color, shrimp connective tissue, shrimp body parameters were found different each other according to the result of the panel. According to the result of the cooked shrimp samples among all properties, cooked odour, flavours, texture were found to be different from each other, as well. Especially, flavours and textural properties of cooked shrimps of the Northern Aegean Sea were higher than the Marmara Sea shrimp. Flesh productivity of Northern Aegean Sea shrimp was found as 46.42 %, while that of the Marmara Sea shrimp was found as 47.74 %.

Keywords: shrimp, biological differences, proximate value, sensory, Parapenaus longirostris, flesh productivity

Procedia PDF Downloads 279
3633 Effect of Plant Density and Planting Pattern on Yield and Quality of Single Cross 704 Silage Corn (Zea mays L.) in Isfahan

Authors: Seyed Mohammad Ali Zahedi

Abstract:

This field experiment was conducted in Isfahan in 2011 in order to study the effect of plant density and planting pattern on growth, yield and quality of silage corn (SC 704) using a randomized complete block design with split plot layout and four replications. The main plot consisted of three planting patterns (60 and 75 cm single planting row and 75 cm double planting row referred to as 60S, 75S and 75T, respectively). The subplots consisted of four levels of plant densities (65000, 80000, 95000 and 110000 plants per hectare). Each subplot consisted of 7 rows, each with 10m length. Vegetative and reproductive characteristics of plants at silking and hard dough stages (when the plants were harvested for silage) were evaluated. Results of variance analysis showed that the effects of planting pattern and plant density were significant on leaf area per plant, leaf area index (at silking), plant height, stem diameter, dry weights of leaf, stem and ear in silking and harvest stages and on fresh and dry yield, dry matter percentage and crude protein percentage at harvest. There was no planting pattern × plant density interaction for these parameters. As row space increased from 60 cm with single planting to 75 cm with single planting, leaf area index and plant height increased, but leaf area per plant, stem diameter, dry weight of leaf, stem and ear, dry matter percentage, dry matter yield and crude protein percentage decreased. Dry matter yield reduced from 24.9 to 18.5 t/ha and crude protein percentage decreased from 6.11 to 5.60 percent. When the plant density increased from 65000 to 110000 plant per hectare, leaf area index, plant height, dry weight of leaf, stem and ear and dry matter yield increased from 19.2 to 23.3 t/ha, whereas leaf area per plant, stem diameter, dry matter percentage and crude protein percentage decreased from 6.30 to 5.25. The best results were obtained with 60 cm row distance with single planting and 110000 plants per hectare.

Keywords: silage corn, plant density, planting pattern, yield

Procedia PDF Downloads 338
3632 Fermented Fruit and Vegetable Discard as a Source of Feeding Ingredients and Functional Additives

Authors: Jone Ibarruri, Mikel Manso, Marta Cebrián

Abstract:

A high amount of food is lost or discarded in the World every year. In addition, in the last decades, an increasing demand of new alternative and sustainable sources of proteins and other valuable compounds is being observed in the food and feeding sectors and, therefore, the use of food by-products as nutrients for these purposes sounds very interesting from the environmental and economical point of view. However, the direct use of discarded fruit and vegetables that present, in general, a low protein content is not interesting as feeding ingredient except if they are used as a source of fiber for ruminants. Especially in the case of aquaculture, several alternatives to the use of fish meal and other vegetable protein sources have been extensively explored due to the scarcity of fish stocks and the unsustainability of fishing for these purposes. Fish mortality is also of great concern in this sector as this problem highly reduces their economic feasibility. So, the development of new functional and natural ingredients that could reduce the need for vaccination is also of great interest. In this work, several fermentation tests were developed at lab scale using a selected mixture of fruit and vegetable discards from a wholesale market located in the Basque Country to increase their protein content and also to produce some bioactive extracts that could be used as additives in aquaculture. Fruit and vegetable mixtures (60/40 ww) were centrifugated for humidity reduction and crushed to 2-5 mm particle size. Samples were inoculated with a selected Rhizopus oryzae strain and fermented for 7 days in controlled conditions (humidity between 65 and 75% and 28ºC) in Petri plates (120 mm) by triplicate. Obtained results indicated that the final fermented product presented a twofold protein content (from 13 to 28% d.w). Fermented product was further processed to determine their possible functionality as a feed additive. Extraction tests were carried out to obtain an ethanolic extract (60:40 ethanol: water, v.v) and remaining biomass that also could present applications in food or feed sectors. The extract presented a polyphenol content of about 27 mg GAE/gr d.w with antioxidant activity of 8.4 mg TEAC/g d.w. Remining biomass is mainly composed of fiber (51%), protein (24%) and fat (10%). Extracts also presented antibacterial activity according to the results obtained in Agar Diffusion and to the Minimum Inhibitory Concentration (MIC) tests determined against several food and fish pathogen strains. In vitro, digestibility was also assessed to obtain preliminary information about the expected effect of extraction procedure on fermented product digestibility. First results indicated that remaining biomass after extraction doesn´t seem to improve digestibility in comparison to the initial fermented product. These preliminary results show that fermented fruit and vegetables can be a useful source of functional ingredients for aquaculture applications and a substitute of other protein sources in the feeding sector. Further validation will be also carried out through “in vivo” tests with trout and bass.

Keywords: fungal solid state fermentation, protein increase, functional extracts, feed ingredients

Procedia PDF Downloads 64
3631 Review of Friction Stir Welding of Dissimilar 5000 and 6000 Series Aluminum Alloy Plates

Authors: K. Subbaiah

Abstract:

Friction stir welding is a solid state welding process. Friction stir welding process eliminates the defects found in fusion welding processes. It is environmentally friend process. 5000 and 6000 series aluminum alloys are widely used in the transportation industries. The Al-Mg-Mn (5000) and Al-Mg-Si (6000) alloys are preferably offer best combination of use in Marine construction. The medium strength and high corrosion resistant 5000 series alloys are the aluminum alloys, which are found maximum utility in the world. In this review, the tool pin profile, process parameters such as hardness, yield strength and tensile strength, and microstructural evolution of friction stir welding of Al-Mg alloys 5000 Series and 6000 series have been discussed.

Keywords: 5000 series and 6000 series Al alloys, friction stir welding, tool pin profile, microstructure and properties

Procedia PDF Downloads 466
3630 Proximate Composition, Colour and Sensory Properties of Akara egbe Prepared from Bambara Groundnut (Vigna subterranea)

Authors: Samson A. Oyeyinka, Taiwo Tijani, Adewumi T. Oyeyinka, Mutiat A. Balogun, Fausat L. Kolawole, John K. Joseph

Abstract:

Bambara groundnut is an underutilised leguminous crop that has a similar composition to cowpea. Hence, it could be used in making traditional snack usually produced from cowpea paste. In this study, akara egbe, a traditional snack was prepared from Bambara groundnut flour or paste. Cowpea was included as the reference sample. The proximate composition and functional properties of the flours were studies as well as the proximate composition and sensory properties of the resulting akara egbe. Protein and carbohydrate were the main components of Bambara groundnut and cowpea grains. Ash, fat and fiber contents were low. Bambara groundnut flour had higher protein content (23.71%) than cowpea (19.47%). In terms of functional properties, the oil absorption capacity (0.75 g oil/g flour) of Bambara groundnut flour was significantly (p ≤ 0.05) lower than that of the cowpea (0.92 g oil/g flour), whereas, Cowpea flour absorbed more water (1.59 g water/g flour) than Bambara groundnut flour (1.12 g/g). The packed bulk density (0.92 g/mL) of Bambara groundnut was significantly (p ≤ 0.05) higher than cowpea flour (0.82 g/mL). Akara egbe prepared from Bambara groundnut flour showed significantly (p ≤ 0.05) higher protein content (23.41%) than the sample made from Bambara groundnut paste (19.35%). Akara egbe prepared from cowpea paste had higher ratings in aroma, colour, taste, crunchiness and overall acceptability than those made from cowpea flour or Bambara groundnut paste or flour. Bambara groundnut can produce akara egbe with comparable nutritional and sensory properties to that made from cowpea.

Keywords: Bambara groundnut, Cowpea, Snack, Sensory properties

Procedia PDF Downloads 264
3629 An Insight into the Paddy Soil Denitrifying Bacteria and Their Relation with Soil Phospholipid Fatty Acid Profile

Authors: Meenakshi Srivastava, A. K. Mishra

Abstract:

This study characterizes the metabolic versatility of denitrifying bacterial communities residing in the paddy soil using the GC-MS based Phospholipid Fatty Acid (PLFA) analyses simultaneously with nosZ gene based PCR-DGGE (Polymerase Chain Reaction-Denaturing Gradient Gel Electrophoresis) and real time Q-PCR analysis. We have analyzed the abundance of nitrous oxide reductase (nosZ) genes, which was subsequently related to soil PLFA profile and DGGE based denitrifier community structure. Soil denitrifying bacterial community comprised majority or dominance of Ochrobactrum sp. following Cupriavidus and uncultured bacteria strains in paddy soil of selected sites. Initially, we have analyzed the abundance of the nitrous oxide reductase gene (nosZ), which was found to be related with PLFA based lipid profile. Chandauli of Eastern UP, India represented greater amount of lipid content (C18-C20) and denitrifier’s diversity. This study suggests the positive co-relation between soil PLFA profiles, DGGE, and Q-PCR data. Thus, a close networking among metabolic abilities and taxonomic composition of soil microbial communities existed, and subsequently, such work at greater extent could be helpful in managing nutrient dynamics as well as microbial dynamics of paddy soil ecosystem.

Keywords: denaturing gradient gel electrophoresis, DGGE, nitrifying and denitrifying bacteria, PLFA, Q-PCR

Procedia PDF Downloads 124
3628 Improvement of Protein Extraction From Shrimp by Product Used for Electrospinning by Applying Emerging Technologies

Authors: Mario Pérez-Won, Vilbett Briones L., Guido Trautmann, María José Bugueño, Gipsy Tabilo-Munizaga, Luis Gonzalez-Cavieres

Abstract:

The fishing industry generates a significant amount of shrimp byproducts, which often result in environmental contamination. Protein extraction from these by-products is a potential solution to minimize waste and revalue the by-products. To improve the extraction of proteins (by chemical method) from shrimp (Pleuroncodes monodon) by-products, the emerging technologies of ohmic heating (OH), microwaves (MW) and pulsed electric fields (PEF) were used. The results show that microwaves, electrical pulses, and ohmic heating improved performance by 28.19%, 19.25%, and 3.65%, respectively. Furthermore, conformational changes were studied by DSC and FTIR. Subsequently, the use of these proteins in electrospinning technology was evaluated. In conclusion, this study demonstrates that the application of emerging technologies, can significantly improve the extraction yield of proteins from shrimp by-products.

Keywords: electrospinning, emerging technologies, improving extraction, shrimp by-products

Procedia PDF Downloads 77
3627 Magnetic Nanoparticles Coated with Modified Polysaccharides for the Immobilization of Glycoproteins

Authors: Kinga Mylkie, Pawel Nowak, Marta Z. Borowska

Abstract:

The most important proteins in human serum responsible for drug binding are human serum albumin (HSA) and α1-acid glycoprotein (AGP). The AGP molecule is a glycoconjugate containing a single polypeptide chain composed of 183 amino acids (the core of the protein), and five glycan branched chains (sugar part) covalently linked by an N-glycosidic bond with aspartyl residues (Asp(N) -15, -38, -54, -75, - 85) of polypeptide chain. This protein plays an important role in binding alkaline drugs, a large group of drugs used in psychiatry, some acid drugs (e.g., coumarin anticoagulants), and neutral drugs (steroid hormones). The main goal of the research was to obtain magnetic nanoparticles coated with biopolymers in a chemically modified form, which will have highly reactive functional groups able to effectively immobilize the glycoprotein (acid α1-glycoprotein) without losing the ability to bind active substances. The first phase of the project involved the chemical modification of biopolymer starch. Modification of starch was carried out by methods of organic synthesis, leading to the preparation of a polymer enriched on its surface with aldehyde groups, which in the next step was coupled with 3-aminophenylboronic acid. Magnetite nanoparticles coated with starch were prepared by in situ co-precipitation and then oxidized with a 1 M sodium periodate solution to form a dialdehyde starch coating. Afterward, the reaction between the magnetite nanoparticles coated with dialdehyde starch and 3-aminophenylboronic acid was carried out. The obtained materials consist of a magnetite core surrounded by a layer of modified polymer, which contains on its surface dihydroxyboryl groups of boronic acids which are capable of binding glycoproteins. Magnetic nanoparticles obtained as carriers for plasma protein immobilization were fully characterized by ATR-FTIR, TEM, SEM, and DLS. The glycoprotein was immobilized on the obtained nanoparticles. The amount of mobilized protein was determined by the Bradford method.

Keywords: glycoproteins, immobilization, magnetic nanoparticles, polysaccharides

Procedia PDF Downloads 130
3626 Nutritional Evaluation of Different Quercus Species in Temperate Regions of Himachal Pradesh

Authors: Ankush Verma, Rohit Bishist

Abstract:

The present investigation was carried out at different locations of Shimla and Kinnaur district and nutrient analysis was done in the laboratory of Department of Silviculture and Agroforestry, Dr. Y.S. Parmar University of Horticulture and Forestry, Nauni, Distt. Solan, Himachal Pradesh during 2019-2020 with the objectives to study the seasonal variation in the nutritive value of different Quercus species and to study the farmers’ preference rating of fodder tress species. From each location leaf samples were collected at 3 months interval from each Quercus spp. The findings of the present study revealed that the nutritional traits of leaves of different Quercus species varied among different seasons throughout the year. The dry matter (61.12 to 64.99%), ether extract (4.07 to 4.42%), crude fibre (34.38 to 37.85%), neutral detergent fibre (57.70 to 61.54%), acid detergent fibre (44.64 to 48.51%), total ash (3.57 to 3.91%), acid insoluble ash (44.64 to 48.51%) and calcium (1.31 to 1.53%) increased with the maturity in the leaves of different Quercus species. While, crude protein (9.10 to 10.61%), nitrogen free extract (44.73 to 47.41%), organic matter (96.09 to 96.43%), and phosphorus (0.16 to 0.31%) decreased with the advancing maturity in the leaves of different Quercus species. Maximum mean values for dry matter (65.05%), ether extract (4.45%), crude fibre (40.82%), neutral detergent fibre (61.48%), acid detergent fibre (48.44%), and organic matter (96.67%) among different Quercus species were recorded in Quercus ilex, while, Maximum mean values for crude protein (10.54%), nitrogen free extract (50.53%), total ash (4.05%), acid insoluble ash (0.59%), calcium (1.61%) and phosphorus (0.40%) were recorded in Quercus leucotrichophora.

Keywords: nutritional evaluation, fodder species, crude protein, carbohydrates

Procedia PDF Downloads 88
3625 Orotic Acid-Induced Fatty Liver in Mink: Characterization and Testing of Bioactive Peptides for Prevention and Treatment

Authors: Don Buddika Oshadi Malaweera, Lora Harris, Bruce Rathgeber, Chibuike C. Udenigwe, Kirsti Rouvinen-Watt

Abstract:

Fatty liver disease is among the three most severe health concerns for mink and believed to occur through the same mechanism as nursing sickness. In North America, nursing sickness affects about 45% of mink farms and in Canada, approximately 50,000 mink females is affected annually. Orotic acid (OA) plays a critical role in lipid metabolism and can increase hepatic lipids by enhancing Sterol regulatory element binding protein-1c expression and decreasing Carnitine palmitoyl transferase I activity. This study was conducted to identify particular pathways and regulatory control points involved in fatty liver development, and evaluate the effectiveness of arginine and bioactive peptides for prevention and treatment of fatty liver disease in mink. A total of 45 mink were used in 9 treatments. The experimental diets consisted of 1% OA, 2% L-arginine and 5% of whey protein hydrolysates. At the end of 10 days of experimental period, the mink were anaesthetized, sampled for blood and euthanized, samples were obtained for histological, biochemical and molecular assays. The blood samples will be analyzed for clinical chemistry and triacylglycerol. The liver samples will be analyzed for total lipid content and analyzed for 6 genes of interest involved in adipogenic transformation, ER stress, and liver inflammation.

Keywords: fatty liver, L-arginine, mink, orotic acid, whey protein hydrolysates

Procedia PDF Downloads 302
3624 Effect of Planting Date on Quantitative and Qualitative Characteristics of Different Bread Wheat and Durum Cultivars

Authors: Mahdi Nasiri Tabrizi, A. Dadkhah, M. Khirkhah

Abstract:

In order to study the effect of planting on yield, yield components and quality traits in bread and durum wheat varieties, a field split-plot experiment based on complete randomized design with three replications was conducted in Agricultural and Natural Resources Research Center of Razavi Khorasan located in city of Mashhad during 2013-2014. Main factor were consisted of five sowing dates (first October, fifteenth December, first March, tenth March, twentieth March) and as sub-factors consisted of different bread wheat (Bahar, Pishgam, Pishtaz, Mihan, Falat and Karim) and two durum wheat (Dena and Dehdasht). According to results of analysis variance the effect of planting date was significant on all examined traits (grain yield, biological yield, harvest index, number of grain per spike, thousands kernel weight, number of spike per square meter, plant height, the number of days to heading, the number of days to maturity, during the grain filling period, percentage of wet gluten, percentage of dry gluten, gluten index, percentage of protein). By delay in planting, majority of traits significantly decreased, except quality traits (percentage of wet gluten, percentage of dry gluten and percentage of protein). Results of means comparison showed, among planting date the highest grain yield and biological yield were related to first planting date (Octobr) with mean of production of 5/6 and 1/17 tons per hectare respectively and the highest bread quality (gluten index) with mean of 85 and percentage of protein with mean of 13% to fifth planting date also the effect of genotype was significant on all traits. The highest grain yield among of studied wheat genotypes was related to Dehdasht cultivar with an average production of 4.4 tons per hectare. The highest protein percentage and bread quality (gluten index) were related to Dehdasht cultivar with 13.4% and Falat cultivar with number of 90 respectively. The interaction between cultivar and planting date was significant on all traits and different varieties had different trend for these traits. The highest grain yield was related to first planting date (October) and Falat cultivar with an average of production of 6/7 tons per hectare while in grain yield did not show a significant different with Pishtas and Mihan cultivars also the most of gluten index (bread quality index) and protein percentage was belonged to the third planting date and Karim cultivar with 7.98 and Dena cultivar with 7.14% respectively.

Keywords: yield component, yield, planting date, cultivar, quality traits, wheat

Procedia PDF Downloads 430
3623 Phylogenetic Study of L1 Protein Human Papillomavirus Type 16 From Cervical Cancer Patients in Bandung

Authors: Fitri Rahmi Fadhilah, Edhyana Sahiratmadja, Ani Melani Maskoen, Ratu Safitri, Supartini Syarif, Herman Susanto

Abstract:

Cervical cancer is the second most common cancer in women after breast cancer. In Indonesia, the incidence of cervical cancer cases is estimated at 25-40 per 100,000 women per year. Human papillomavirus (HPV) infection is a major cause of cervical cancer, and HPV-16 is the most common genotype that infects the cervical tissue. The major late protein L1 may be associated with infectivity and pathogenicity and its variation can be used to classify HPV isolates. The aim of this study was to determine the phylogenetic tree of HPV 16 L1 gene from cervical cancer patient isolates in Bandung. After confirming HPV-16 by Linear Array Genotyping Test, L1 gene was amplified using specific primers and subject for sequencing. Phylogenetic analysis revealed that HPV 16 from Bandung was in the subgroup of Asia and East Asia, showing the close host-agent relationship among the Asian type.

Keywords: L1 HPV 16, cervical cancer, bandung, phylogenetic

Procedia PDF Downloads 503
3622 Sensitivity Analysis for 14 Bus Systems in a Distribution Network with Distributed Generators

Authors: Lakshya Bhat, Anubhav Shrivastava, Shiva Rudraswamy

Abstract:

There has been a formidable interest in the area of Distributed Generation in recent times. A wide number of loads are addressed by Distributed Generators and have better efficiency too. The major disadvantage in Distributed Generation is voltage control- is highlighted in this paper. The paper addresses voltage control at buses in IEEE 14 Bus system by regulating reactive power. An analysis is carried out by selecting the most optimum location in placing the Distributed Generators through load flow analysis and seeing where the voltage profile rises. MATLAB programming is used for simulation of voltage profile in the respective buses after introduction of DG’s. A tolerance limit of +/-5% of the base value has to be maintained. To maintain the tolerance limit, 3 methods are used. Sensitivity analysis of 3 methods for voltage control is carried out to determine the priority among the methods.

Keywords: distributed generators, distributed system, reactive power, voltage control, sensitivity analysis

Procedia PDF Downloads 703
3621 Nutritional Composition of Crackers Produced from Blend of Sprouted Pigeon Pea (Cajanus cajan), Unripe Plantain (Musa parasidiaca), and Brewers’ Spent Grain Flour and Blood Glucose Level of Diabetic Rats Fed the Biscuit

Authors: Nneka N. Uchegbu, Charles N. Ishiwu

Abstract:

The nutritional composition and hypoglycaemic effect of crackers produced from a blend of sprouted pigeon pea, unripe plantain, and brewers’ spent grain and fed to Alloxan induced diabetic rat was investigated. Crackers were produced from different blends of sprouted pigeon pea, unripe plantain and brewers’ spent grain. The crackers were evaluated for proximate composition, amino acid profile and antinutritional factors. Blood glucose levels of normal and diabetic rats fed with the control sample and different formulations of cracker were measured. The protein content of the samples were significantly different (p < 0.05) from each other with sample A having the lowest value and sample B with the highest value. The values obtained showed that the samples contained most of the amino acids that are found in plant proteins. The levels of antinutritional factor determined were generally low. Administration of the formulated cracker meals led to a significant reduction in the fasting blood glucose level in the diabetic rats. The present study concluded that consumption of crackers produced from this composite flour can be recommended for the diabetics and those who are sceptical about the disease.

Keywords: crackers, diabetics rat, sprouted pigeon pea, unripe plantain and brewers’ spent grain

Procedia PDF Downloads 440
3620 Effect of 10 Weeks of Aerobic Exercise Training on Serum Concentrations of Surfactant Protein D and Insulin Resistance in Women with Type 2 Diabetes

Authors: Sajjad Rezaei, Mahdieh Molanouri Shamsi, Azadeh Jamali

Abstract:

Background and purpose: Surfactant protein D (SP-D) is a lung-specific protein that is detectable in human plasma. Effect of exercise training on SP-D levels as well as its relation to metabolic indices is not known. The present study then aimed to investigate the effects of 10 weeks of aerobic training on serum levels of SP-D and insulin resistance in women with type 2 diabetes. Materials and methods: Twenty-two overweight women with type 2 diabetes mellitus were recruited through deliberate sampling and randomly assigned to intervention and control groups (11 in each group). The intervention group underwent a progressive aerobic training program for 10 weeks, 3 days per week, 30-55 min/day at 50-75% heart rate reserve (HRR). Control group continued with its everyday routine. Blood samples were obtained before and after training for biochemical analysis. Within-group and between-group differences were analyzed with paired and independent t-tests in spss software, respectively, and the relation between variables was analyzed with Pearson’s correlation coefficient (all at P = 0.05). Results: Significant differences were observed between groups in leptin, glucose, waist circumference and VO2 max after training. SP-D was decreased and VO2 max was increased significantly in intervention group. However, no significant correlation was observed between SP-D and other variables. Conclusion: Since there was no corresponding decrease in insulin resistance with decreased levels of SP-D, it seems unlikely for SP-D to mediate the association between obesity and insulin resistance in type 2 diabetics.

Keywords: exercise training, SP-D, insulin resistance, type 2 diabetes

Procedia PDF Downloads 416
3619 Differential Proteomic Profile and Terpenoid Production in Somatic Embryos of Jatropha curcas

Authors: Anamarel Medina-Hernandez, Teresa Ponce-Noyola, Ileana Vera-Reyes, Ana C. Ramos-Valdivia

Abstract:

Somatic embryos reproduce original seed characteristics and could be implemented in biotechnological studies. Jatropha curcas L. is an important plant for biodiesel production, but also is used in traditional medicine. Seeds from J. curcas are toxic because contain diterpenoids called phorbol esters, but in Mexico exist a non-toxic variety. Therefore, somatic embryos suspension cultures from non-toxic J. curcas variety were induced. In order to investigate the characteristics of somatic embryos, a differential proteomic analysis was made between pre-globular and globular stages by 2-D gel electrophoresis. 108 spots were differentially expressed (p<0.02), and 20 spots from globular somatic embryos were sequenced by MALDI-TOF-TOF mass spectrometry. A comparative analysis of terpenoids production between the two stages was made by RP-18 TLC plates. The sequenced proteins were related to energy production (68%), protein destination and storage (9%), secondary metabolism (9%), signal transduction (5%), cell structure (5%) and aminoacid metabolism (4%). Regarding terpenoid production, in pre-globular and globular somatic embryos were identified sterols and triterpenes of pharmacological interest (alpha-amyrin and betulinic acid) but also it was found compounds that were unique to each stage. The results of this work are the basis to characterize at different levels the J. curcas somatic embryos so that this system can be used efficiently in biotechnological processes.

Keywords: Jatropha curcas, proteomics, somatic embryo, terpenoids

Procedia PDF Downloads 256
3618 Links between Inflammation and Insulin Resistance in Children with Morbid Obesity and Metabolic Syndrome

Authors: Mustafa M. Donma, Orkide Donma

Abstract:

Obesity is a clinical state associated with low-grade inflammation. It is also a major risk factor for insulin resistance (IR). In its advanced stages, metabolic syndrome (MetS), a much more complicated disease which may lead to life-threatening problems, may develop. Obesity-mediated IR seems to correlate with the inflammation. Human studies performed particularly on pediatric population are scarce. The aim of this study is to detect possible associations between inflammation and IR in terms of some related ratios. 549 children were grouped according to their age- and sex-based body mass index (BMI) percentile tables of WHO. MetS components were determined. Informed consent and approval from the Ethics Committee for Clinical Investigations were obtained. The principles of the Declaration of Helsinki were followed. The exclusion criteria were infection, inflammation, chronic diseases and those under drug treatment. Anthropometric measurements were obtained. Complete blood cell, fasting blood glucose, insulin, and C-reactive protein (CRP) analyses were performed. Homeostasis model assessment of insulin resistance (HOMA-IR), systemic immune inflammation (SII) index, tense index, alanine aminotransferase to aspartate aminotransferase ratio (ALT/AST), neutrophils to lymphocyte (NLR), platelet to lymphocyte, and lymphocyte to monocyte ratios were calculated. Data were evaluated by statistical analyses. The degree for statistical significance was 0.05. Statistically significant differences were found among the BMI values of the groups (p < 0.001). Strong correlations were detected between the BMI and waist circumference (WC) values in all groups. Tense index values were also correlated with both BMI and WC values in all groups except overweight (OW) children. SII index values of children with normal BMI were significantly different from the values obtained in OW, obese, morbid obese and MetS groups. Among all the other lymphocyte ratios, NLR exhibited a similar profile. Both HOMA-IR and ALT/AST values displayed an increasing profile from N towards MetS3 group. BMI and WC values were correlated with HOMA-IR and ALT/AST. Both in morbid obese and MetS groups, significant correlations between CRP versus SII index as well as HOMA-IR versus ALT/AST were found. ALT/AST and HOMA-IR values were correlated with NLR in morbid obese group and with SII index in MetS group, (p < 0.05), respectively. In conclusion, these findings showed that some parameters may exhibit informative differences between the early and late stages of obesity. Important associations among HOMA-IR, ALT/AST, NLR and SII index have come to light in the morbid obese and MetS groups. This study introduced the SII index and NLR as important inflammatory markers for the discrimination of normal and obese children. Interesting links were observed between inflammation and IR in morbid obese children and those with MetS, both being late stages of obesity.

Keywords: children, inflammation, insulin resistance, metabolic syndrome, obesity

Procedia PDF Downloads 137
3617 Safety Profile of Anti-Retroviral Medicine in South Africa Based on Reported Adverse Drug Reactions

Authors: Sarah Gounden, Mukesh Dheda, Boikhutso Tlou, Elizabeth Ojewole, Frasia Oosthuizen

Abstract:

Background: Antiretroviral therapy (ART) has been effective in the reduction of mortality and resulted in an improvement in the prognosis of HIV-infected patients. However, treatment with antiretrovirals (ARVs) has led to the development of many adverse drug reactions (ADRs). It is, therefore, necessary to determine the safety profile of these medicines in a South African population in order to ensure safe and optimal medicine use. Objectives: The aim of this study was to quantify ADRs experienced with the different ARVs currently used in South Africa, to determine the safety profile of ARV medicine in South Africa based on reported ADRs, and to determine the ARVs with the lowest risk profile based on specific patient populations. Methodology: This was a quantitative study. Individual case safety reports for the period January 2010 – December 2013 were obtained from the National Pharmacovigilance Center; these reports contained information on ADRs, ARV medicine, and patient demographics. Data was analysed to find associations that may exist between ADRs experienced, ARV medicines used and patient demographics. Results: A total of 1916 patient reports were received of which 1534 met the inclusion criteria for the study. The ARV with the lowest risk of ADRs were found to be lamivudine (0.51%, n=12), followed by lopinavir/ritonavir combination (0.8%, n=19) and abacavir (0.64%, n=15). A higher incidence of ADRs was observed in females compared to males. The age group 31–50 years and the weight group 61–80 kg had the highest incidence of ADRs reported. Conclusion: This study found that the safest ARVs to be used in a South African population are lamivudine, abacavir, and the lopinavir/ritonavir combination. Gender differences play a significant role in the occurrence of ADRs and both anatomical and physiological differences account for this. An increased BMI (body mass index) in both men and women showed an increase in the incidence of ADRs associated with ARV therapy.

Keywords: adverse drug reaction, antiretrovirals, HIV/AIDS, pharmacovigilance, South Africa

Procedia PDF Downloads 351
3616 Sensitivity Analysis for 14 Bus Systems in a Distribution Network with Distribution Generators

Authors: Lakshya Bhat, Anubhav Shrivastava, Shivarudraswamy

Abstract:

There has been a formidable interest in the area of Distributed Generation in recent times. A wide number of loads are addressed by Distributed Generators and have better efficiency too. The major disadvantage in Distributed Generation is voltage control- is highlighted in this paper. The paper addresses voltage control at buses in IEEE 14 Bus system by regulating reactive power. An analysis is carried out by selecting the most optimum location in placing the Distributed Generators through load flow analysis and seeing where the voltage profile rises. Matlab programming is used for simulation of voltage profile in the respective buses after introduction of DG’s. A tolerance limit of +/-5% of the base value has to be maintained.To maintain the tolerance limit , 3 methods are used. Sensitivity analysis of 3 methods for voltage control is carried out to determine the priority among the methods.

Keywords: distributed generators, distributed system, reactive power, voltage control, sensitivity analysis

Procedia PDF Downloads 587
3615 Modified Acetamidobenzoxazolone Based Biomarker for Translocator Protein Mapping during Neuroinflammation

Authors: Anjani Kumar Tiwari, Neelam Kumari, Anil Mishra

Abstract:

The 18-kDa translocator protein (TSPO) previously called as peripheral benzodiazepine receptor, is proven biomarker for variety of neuroinflammation. TSPO is tryptophane rich five transmembranal protein found on outer mitochondrial membrane of steroid synthesising and immunomodulatory cells. In case of neuronal damage or inflammation the expression level of TSPO get upregulated as an immunomodulatory response. By utilizing Benzoxazolone as a basic scaffold, series of TSPO ligands have been designed followed by their screening through in silico studies. Synthesis has been planned by employing convergent methodology in six high yielding steps. For the synthesized ligands the ‘in vitro’ assay was performed to determine the binding affinity in term of Ki. On ischemic rat brain, autoradiography studies were also carried to check the specificity and affinity of the designed radiolabelled ligand for TSPO.Screening was performed on the basis of GScore of CADD based schrodinger software. All the modified and better prospective compound were successfully carried out and characterized by spectroscopic techniques (FTIR, NMR and HRMS). In vitro binding assay showed best binding affinity Ki = 6.1+ 0.3 for TSPO over central benzodiazepine receptor (CBR) Ki > 200. ARG studies indicated higher uptake of two analogues on the lesion side compared with that on the non-lesion side of ischemic rat brains. Displacement experiments with unlabelled ligand had minimized the difference in uptake between the two sides which indicates the specificity of the ligand towards TSPO receptor.

Keywords: TSPO, PET, imaging, Acetamidobenzoxazolone

Procedia PDF Downloads 143
3614 The Effects of pH on p53 Phosphorylation by Ataxia Telangiectasia Mutated Kinase

Authors: Serap Pektas

Abstract:

Ataxia telangiectasia mutated (ATM) is a serine-threonine kinase, which is the major regulator of the DNA damage response. ATM is activated upon the formation of DNA double-strand breaks (DSBs) in the cells. ATM phosphorylates the proteins involved in apoptotic responses, cell cycle checkpoint control, DNA repair, etc. Tumor protein p53, known as p53 is one of these proteins that phosphorylated by ATM. Phosphorylation of p53 at Ser15 residue leads to p53 stabilization in the cells. Often enzymes activity is affected by hydrogen ion concentration (pH). In order to find the optimal pH range for ATM activity, steady-state kinetic assays were performed at acidic and basic pH ranges. Ser15 phosphorylation of p53 is determined by using ELISA. The results indicated that the phosphorylation rate was better at basic pH range compared with the acidic pH range. This could be due to enzyme stability, or enzyme-substrate interaction is pH dependent.

Keywords: ataxia telangiectasia mutated, DNA double strand breaks, DNA repair, tumor protein p53

Procedia PDF Downloads 134
3613 Determination of Nutritional Value and Steroidal Saponin of Fenugreek Genotypes

Authors: Anita Singh, Richa Naula, Manoj Raghav

Abstract:

Nutrient rich and high-yielding varieties of fenugreek can be developed by using genotypes which are naturally high in nutrients. Gene banks harbour scanty germplasm collection of Trigonella spp. and a very little background information about its genetic diversity. The extent of genetic diversity in a specific breeding population depends upon the genotype included in it. The present investigation aims at the estimation of macronutrient (phosphorus by spectrophotometer and potassium by flame photometer), micronutrients, namely, iron, zinc, manganese, and copper from seeds of fenugreek genotypes using atomic absorption spectrophotometer, protein by Rapid N Cube Analyser and Steroidal Saponins. Twenty-eight genotypes of fenugreek along with two standard checks, namely, Pant Ragini and Pusa Early Bunching were collected from different parts of India, and nutrient contents of each genotype were determined at G. B. P. U. A. & T. Laboratory, Pantnagar. Highest potassium content was observed in PFG-35 (1207 mg/100g). PFG-37 and PFG-20 were richest in phosphorus, iron and manganese content among all the genotypes. The lowest zinc content was found in PFG-26 (1.19 mg/100g), while the maximum zinc content was found in PFG- 28 (4.43 mg/100g). The highest content of copper was found in PFG-26 (1.97 mg/100g). PFG-39 has the highest protein content (29.60 %). Significant differences were observed in the steroidal saponin among the genotypes. Saponin content ranged from 0.38 g/100g to 1.31 g/100g. Steroidal Saponins content was found the maximum in PFG-36 (1.31 g/100g) followed by PFG-17 (1.28 g/100g). Therefore, the genotypes which are rich in nutrient and oil content can be used for plant biofortification, dietary supplements, and herbal products.

Keywords: genotypes, macronutrients, micronutrient, protein, seeds

Procedia PDF Downloads 254
3612 Development of an Integrated Route Information Management Software

Authors: Oluibukun G. Ajayi, Joseph O. Odumosu, Oladimeji T. Babafemi, Azeez Z. Opeyemi, Asaleye O. Samuel

Abstract:

The need for the complete automation of every procedure of surveying and most especially, its engineering applications cannot be overemphasized due to the many demerits of the conventional manual or analogue approach. This paper presents the summarized details of the development of a Route Information Management (RIM) software. The software, codenamed ‘AutoROUTE’, was encoded using Microsoft visual studio-visual basic package, and it offers complete automation of the computational procedures and plan production involved in route surveying. It was experimented using a route survey data (longitudinal profile and cross sections) of a 2.7 km road which stretches from Dama to Lunko village in Minna, Niger State, acquired with the aid of a Hi-Target DGPS receiver. The developed software (AutoROUTE) is capable of computing the various simple curve parameters, horizontal curve, and vertical curve, and it can also plot road alignment, longitudinal profile, and cross-section with a capability to store this on the SQL incorporated into the Microsoft visual basic software. The plotted plans with AutoROUTE were compared with the plans produced with the conventional AutoCAD Civil 3D software, and AutoROUTE proved to be more user-friendly and accurate because it plots in three decimal places whereas AutoCAD plots in two decimal places. Also, it was discovered that AutoROUTE software is faster in plotting and the stages involved is less cumbersome compared to AutoCAD Civil 3D software.

Keywords: automated systems, cross sections, curves, engineering construction, longitudinal profile, route surveying

Procedia PDF Downloads 148
3611 Qualitative Characterization of Proteins in Common and Quality Protein Maize Corn by Mass Spectrometry

Authors: Benito Minjarez, Jesse Haramati, Yury Rodriguez-Yanez, Florencio Recendiz-Hurtado, Juan-Pedro Luna-Arias, Salvador Mena-Munguia

Abstract:

During the last decades, the world has experienced a rapid industrialization and an expanding economy favoring a demographic boom. As a consequence, countries around the world have focused on developing new strategies related to the production of different farm products in order to meet future demands. Consequently, different strategies have been developed seeking to improve the major food products for both humans and livestock. Corn, after wheat and rice, is the third most important crop globally and is the primary food source for both humans and livestock in many regions around the globe. In addition, maize (Zea mays) is an important source of protein accounting for up to 60% of the daily human protein supply. Generally, many of the cereal grains have proteins with relatively low nutritional value, when they are compared with proteins from meat. In the case of corn, much of the protein is found in the endosperm (75 to 85%) and is deficient in two essential amino acids, lysine, and tryptophan. This deficiency results in an imbalance of amino acids and low protein content; normal maize varieties have less than half of the recommended amino acids for human nutrition. In addition, studies have shown that this deficiency has been associated with symptoms of growth impairment, anemia, hypoproteinemia, and fatty liver. Due to the fact that most of the presently available maize varieties do not contain the quality and quantity of proteins necessary for a balanced diet, different countries have focused on the research of quality protein maize (QPM). Researchers have characterized QPM noting that these varieties may contain between 70 to 100% more residues of the amino acids essential for animal and human nutrition, lysine, and tryptophan, than common corn. Several countries in Africa, Latin America, as well as China, have incorporated QPM in their agricultural development plan. Large parts of these countries have chosen a specific QPM variety based on their local needs and climate. Reviews have described the breeding methods of maize and have revealed the lack of studies on genetic and proteomic diversity of proteins in QPM varieties, and their genetic relationships with normal maize varieties. Therefore, molecular marker identification using tools such as mass spectrometry may accelerate the selection of plants that carry the desired proteins with high lysine and tryptophan concentration. To date, QPM maize lines have played a very important role in alleviating the malnutrition, and better characterization of these lines would provide a valuable nutritional enhancement for use in the resource-poor regions of the world. Thus, the objectives of this study were to identify proteins in QPM maize in comparison with a common maize line as a control.

Keywords: corn, mass spectrometry, QPM, tryptophan

Procedia PDF Downloads 288
3610 Plasma-Induced Modification of Biomolecules: A Tool for Analysis of Protein Structures

Authors: Yuting Wu, Faraz Choudhury, Daniel Benjamin, James Whalin, Joshua Blatz, Leon Shohet, Michael Sussman, Mark Richards

Abstract:

Plasma-Induced Modification of Biomolecules (PLIMB) has been developed as a technology, which, together with mass spectrometry, measures three-dimensional structural characteristics of proteins. This technique uses hydroxyl radicals generated by atmospheric-pressure plasma discharge to react with the solvent-accessible side chains of protein in an aqueous solution. In this work, we investigate the three-dimensional structure of hemoglobin and myoglobin using PLIMB. Additional modifications to these proteins, such as oxidation, fragmentations, and conformational changes caused by PLIMB are also explored. These results show that PLIMB, coupled with mass spectrometry, is an effective way to determine solvent access to hemoproteins. Furthermore, we show that many factors, including pH and the electrical parameters used to generate the plasma, have a significant influence on solvent accessibility.

Keywords: plasma, hemoglobin, myoglobin, solvent access

Procedia PDF Downloads 193
3609 Aspergillus micromycetes as Producers of Hemostatically Active Proteases

Authors: Alexander A. Osmolovskiy, Anastasia V. Orekhova, Daria M. Bednenko, Yelyzaveta Boiko

Abstract:

Micromycetes from Aspergillus genus can produce proteases capable of promoting proteolysis of hemostasis proteins or, along with hydrolytic activity, to show the ability to convert proenzymes of this system activating them into an active form. At the same time, practical medicine needs specific activators for quantitation of the level of some plasma enzymes, especially protein C and factor X, the lack of which leads to the development of thromboembolic diseases. Thus, some micromycetes of the genus Aspergillus were screened for the ability to synthesize extracellular proteases with promising activity for designing anti-thrombotic and diagnostic preparations. Such standard methods like salting out, electrophoresis, isoelectrofocusing were used for isolation, purification and study of physicochemical properties of proteases. Enzyme activity was measured spectrophotometrically fibrin as a substrate of the reaction and chromogenic peptide substrates of different proteases of the human hemostasis system. As a result of the screening, four active producers were selected: Aspergillus janus 301, A. flavus 1, A. terreus 2, and A. ochraceus L-1. The enzyme of A. janus 301 showed the greatest fibrinolytic activity (around 329.2 μmol Tyr/(ml × min)). The protease produced by A. terreus 2 had the highest plasmin-like activity (54.1 nmol pNA/(ml × min)), but fibrinolytic activity was lower than A. janus 301 demonstrated (25.2 μmol Tyr/(ml × min)). For extracellular protease of micromycete A. flavus a high plasmin-like activity was also shown (39.8 nmol pNA / (ml × min)). Moreover, according to our results proteases one of the fungi - A. terreus 2 were able to activate protein C of human plasma - the key factor of the human anticoagulant hemostasis system. This type of activity was 39.8 nmol pNA/(ml × min)). It was also shown that A. ochraceus L-1 could produce extracellular proteases with protein C and factor X activator activities (65.9 nmol pNA/(ml × min) and 34.6 nmol pNA/(ml × min) respectively). The maximum accumulation of the proteases falls on the 4th day of cultivation. Using isoelectrofocusing was demonstrated that the activation of both proenzymes might proceed via limited proteolysis induced by proteases of A. ochraceus L-1. The activatory activity of A. ochraceus L-1 proteases toward essential hemostatic proenzymes, protein C and X factor may be useful for practical needs. It is well known that similar enzymes, activators of protein C and X factor isolated from snake venom, South American copperhead Agkistrodon contortrix contortrix and Russell’s viper Daboia russelli russeli, respectively, are used for the in vitro diagnostics of the functional state of these proteins in blood plasma. Thus, the proteases of Aspergillus genus can be used as cheap components for enzyme thrombolytic preparations.

Keywords: anti-trombotic drugs, fibrinolysis, diagnostics, proteases, micromycetes

Procedia PDF Downloads 134