Search results for: scaffold protein

Authors: Ž. Vaštag, Lj. Popović, S. Popović

Abstract:

After cold pressing of pumpkin oil, the defatted oil cake (PUOC) was utilized as raw material for processing of bio-functional hydrolysates. In this study, the in vitro bioactivity of an alcalase (AH) and a pepsin hydrolysate (PH) prepared from the major pumpkin 12S globulin (cucurbitin) are compared. The hydrolysates were produced at optimum reaction conditions (temperature, pH) for the enzymes, during 60min. The bioactivity testing included antioxidant and angiotensin I converting enzyme inhibitory activity assays. The hydrolysates showed high potential as natural antioxidants and possibly antihypertensive agents in functional food or nutraceuticals. Additionally, preliminary studies have shown that both hydrolysates could exhibit modest α-amylase inhibitory activity, which indicates on their hypoglycemic potential.

Keywords: cucurbitin, alcalase, pepsin, protein hydrolysates, in vitro bioactivity

Procedia PDF Downloads 311

Authors: Dong-An Wang

Abstract:

All kinds of microspheres have been extensively employed as carriers for drug, gene and therapeutic cell delivery. Most therapeutic cell delivery microspheres rely on a two-step methodology: fabrication of microspheres and subsequent seeding of cells onto them. In this study, we have developed a novel one-step cell encapsulation technique using a convenient and instant water-in-oil single emulsion approach to form cell-encapsulated gelatin microspheres. This technology is adopted for hyaline cartilage tissue engineering, in which autologous chondrocytes are used as therapeutic cells. Cell viability was maintained throughout and after the microsphere formation (75-100 µm diameters) process that avoids involvement of any covalent bonding reactions or exposure to any further chemicals. Further encapsulation of cell-laden microspheres in alginate gels were performed under 4°C via a prompt process. Upon the formation of alginate constructs, they were immediately relocated into CO2 incubator where the temperature was maintained at 37°C; under this temperature, the cell-laden gelatin microspheres dissolved within hours to yield similarly sized cavities and the chondrocytes were therefore suspended within the cavities inside the alginate gel bulk. Hence, the gelatin cell-laden microspheres served two roles: as cell delivery vehicles which can be removable through temperature curing, and as porogens within an alginate hydrogel construct to provide living space for cell growth and tissue development as well as better permeability for mutual diffusions. These cell-laden microspheres, namely “temperature-cured dissolvable gelatin microsphere based cell carriers” (tDGMCs), were further encapsulated in a chondrocyte-laden alginate scaffold system and analyzed by WST-1, gene expression analyses, biochemical assays, histology and immunochemistry stains. The positive results consistently demonstrated the promise of tDGMC technology in delivering these non-anchorage dependent cells (chondrocytes). It can be further conveniently translated into delivery of other non-anchorage dependent cell species, including stem cells, progenitors or iPS cells, for regeneration of tissues in internal organs, such as engineered hepatogenesis or pancreatic regeneration.

Keywords: biomaterials, tissue engineering, microsphere, hydrogel, porogen, anchorage dependence

Procedia PDF Downloads 396

Authors: Asma Bashir, Neha Farid, Kashif Ali, Kiran Fatima

Abstract:

Background: Bacteriocins are a subclass of antimicrobial peptides that are becoming extremely important in treatments. It is possible to utilise bacteriocins in place of or in addition to traditional antibiotics. It is possible to treat a variety of infections, including Vancomycin-Resistant Staphylococcus aureus (VRSA) and Methicillin-Resistant Staphylococcus aureus (MRSA), using the targeted spectrum of activity of these microorganisms. Method: This study aimed to examine the efficiency of antibiotics and bacteriocin against VRSA and MRSA. The effects of bacteriocins, such as enterocin KAE01, enterocin KAE03, enterocin KAE05, and enterocin KAE06 isolated from Enterococcus faecium strains, alone and in combination with vancomycin and methicillin antibiotics were examined. The selection technique utilized the minimum inhibitory concentrations (MICs) against Gram-positive indicator strain ATCC 6538 Methicillin-Resistant Staphylococcus aureus (MRSA) and indicator strain KSA 02 Vancomycin-Resistant Staphylococcus aureus (VRSA). Results: We report the isolation and identification of enterocins KAE01, KAE03, KAE05, and KAE06 from food isolates of Enterococcus faecium (KAE01, KAE03, KAE05, and KAE06). After isolating the protein, it was partially purified with ammonium sulphate precipitation and purified with fast protein liquid chromatography (FPLC) procedures. Combinations of enterocin KAE01, 1 citric acid, 1 lactic acid, and microcin J25, 1 reuterin, 1 citric acid, and microcin J25, 1 reuterin, 1 lactic acid shown synergistic benefits (FIC index = 0.5) against Vancomycin-Resistant Staphylococcus aureus (VRSA). In addition, a moderately synergistic (FIC index = 0.75) interaction was seen between pediocin PA-1, 1 citric acid, 1 lactic acid, and reuterin 1 citric acid, 1 lactic acid against L. ivanovii HPB28. In the presence of acids, nisin Z exhibited a modestly synergistic effect (FIC index = 0.625-0.75); however, it exhibited additive effects (FIC index = 1) when combined with reuterin or pediocin PA-1 against L. ivanovii HPB28. The efficacy of synergistic consortiums against Gram-positive bacteria was examined. Conclusion: Combining antimicrobials with various modes of action boosted efficacy and expanded the spectrum of inhibition, particularly against multidrug-resistant pathogens, according to our research.

Keywords: Enterococcus faecium, bacteriocin, antimicrobial resistance, antagonistic activity, vancomycin-resistant Staphylococcus aureus, methicillin-resistant Staphylococcus aureus

Procedia PDF Downloads 148

Authors: Hatice Akbıyık, Hülya Gökmen Özel, Nagehan Emiralioğlu, Elmas Ebru Güneş Yalçın, Deniz Doğru Ersöz, Hayriye Uğur Özçelik, Nural Kiper

Abstract:

Background: Dietary recommendations in cystic fibrosis (CF) are based on the need to compensate for the increased energy needs of infection, the increased energy cost of breathing and the losses, incurred from malabsorption. Studies in CF indicate that dietary recommendations for CF patients can be difficult to achieve Aim: The aim of this study was to evaluate the energy and nutrient intakes and to compare in accordance with CF dietary guidelines in CF. Methods: One-hundred sixty patients with CF, aged between 2 to 20 years (mean±SD= 7.4±4.8 years) attending Hacettepe University, Faculty of Medicine, Department of Pediatric Pulmonary Diseases were included. Energy and nutrient intakes from foods and enteral products were calculated using a-24-hour dietary recall method with BEBIS 7.2 programme. Percentages of energy and nutrient intakes were compared in accordance with CF dietary guidelines. Patients or/and parents completed a questionnaire showing mealtime problems, usage of alternative therapies and type of nutrition. Statistical analyses were done using SPSS 16.0 programme. Results: It was obtained that 14.5% and 46.9% of the total energy intake were from proteins and carbohydrates, respectively. The actual contribution of total, saturated, monounsaturated and polyunsaturated fats to the total caloric intake was 37.5%, 14.3%, 14.9%, 9.9%, respectively. It was found that 87.7% of energy, 85% of protein 91.7% of carbohydrate, 81.1% of fat intakes were met, when compared CF recommended intakes of 120% RDA. Additionally 67%, 69.5%, 68.2% and 68.9% of the subjects did not achieve CF recommended intakes of 120% RDA for energy, protein, carbohydrate and fat, respectively. Patients with CF had low intakes for age for almost all vitamins and minerals, although supplementation was given. Especially most patients did not achieve the minimum recommended vitamin K intake of 120% RDA. The percentage meeting 120% RDA was 75.9% for vitamin K. It was shown that 41% of the patients had mealtime problems and they skipped the breakfast. Moreover 25.4% of the patients used alternative products outside the standard treatment (such as omega-3, ginger, turmeric, local honey). It was also showed that 60.8% of patients were using enteral products in addition to normal foods, the remaining patients were on only normal foods. Conclusion: The aims of improving nutritional status in children are to achieve normal weight gain and growth; optimize vitamin and mineral status; and slow the rate of clinical decline. In this study although enteral products were used in patients with CF, it was found that energy and nutrient requirements were unable to meet. Because dietary assessment is essential to identify the need for earlier nutritional intervention, in each visit patients need to be referred to CF specialist dietitian.

Keywords: cystic fibrosis, energy and nutrient intakes, mealtime problems, malabsorbtion

Procedia PDF Downloads 459

Authors: Gaweł Magdalena, Lipkowska Anna, Olbert Magdalena, Frąckiewicz Ewelina, Librowski Tadeusz, Nowak Gabriel, Pilc Andrzej

Abstract:

Heme oxygenase-1 (HO-1), also known as heat shock protein 32 (HSP32), has been shown to be implicated in cytoprotection in various organs. Its activation plays a significant role in acute and chronic inflammation, protecting cells from oxidative injury and apoptosis. This inducible isoform of HO catalyzes the first and rate-limiting step in heme degradation to produce equimolar quantities of biologically active products: carbon monoxide (CO), free iron and biliverdin. CO has been reported to possess anti-apoptotic properties. Moreover, it inhibits the production of proinflammatory cytokines and stimulates the synthesis of the anti-inflammatory interleukin-10 (IL-10), as well as promotes vasodilatation at sites of inflammation. The second product of catalytic HO-1 activity, free cytotoxic iron, is promptly sequestered into the iron storage protein ferritin, which lowers the pro-oxidant state of the cell. The third product, biliverdin, is subsequently converted by biliverdin reductase into the bile pigment bilirubin, the most potent endogenous antioxidant among the constituents of human serum, which modulates immune effector functions and suppresses inflammatory response. Furthermore, being one of the so-called stress proteins, HO-1 adaptively responds to different stressors, such as reactive oxygen species (ROS), inflammatory cytokines and heavy metals and thus protects cells against such conditions as ischemia, hemorrhagic shock, heat shock or hypoxia. It is suggested that pharmacologic modulation of HO-1 may represent an effective strategy for prevention of stress and drug-induced gastrointestinal toxicity. HO-1 is constitutively expressed in normal gastric, intestinal and colonic mucosa and up-regulated during inflammation. It has been proven that HO-1 up-regulated by hemin, heme and cobalt-protoporphyrin ameliorates experimental colitis. In addition, the up-regulation of HO-1 partially explains the mechanism of action of 5-aminosalicylic acid (5-ASA), which is used clinically as an anti-colitis agent. In 2009 Ueda et al. has reported for the first time that mucosal protection by Polaprezinc, a chelate compound of zinc and L-carnosine used as an anti-ulcer drug in Japan, is also attributed to induction of HO-1 in the stomach. Since then, inducers of HO-1 are desired subject of research, as they may constitute therapeutically effective anti-ulcer drugs.

Keywords: heme oxygenase-1, gastric lesions, gastroprotection, Polaprezinc

Procedia PDF Downloads 504

Authors: Zhaoyang Liu

Abstract:

It has been highly desired to develop a high-performance membrane for separating oil/water emulsions with the combined features of high water flux, high oil separation efficiency, and high mechanical stability. Here, we demonstrated a design for high-performance membranes constructed with ultra-long titanate nanofibers (over 30 µm in length)/cellulose microfibers. An integrated network membrane was achieved with these ultra-long nano/microfibers, contrast to the non-integrated membrane constructed with carbon nanotubes (5 µm in length)/cellulose microfibers. The morphological properties of the prepared membranes were characterized by A FEI Quanta 400 (Hillsboro, OR, United States) environmental scanning electron microscope (ESEM). The hydrophilicity, underwater oleophobicity and oil adhesion property of the membranes were examined using an advanced goniometer (Rame-hart model 500, Succasunna, NJ, USA). More specifically, the hydrophilicity of membranes was investigated by analyzing the spreading process of water into membranes. A filtration device (Nalgene 300-4050, Rochester, NY, USA) with an effective membrane area of 11.3 cm² was used for evaluating the separation properties of the fabricated membranes. The prepared oil-in-water emulsions were poured into the filtration device. The separation process was driven under vacuum with a constant pressure of 5 kPa. The filtrate was collected, and the oil content in water was detected by a Shimadzu total organic carbon (TOC) analyzer (Nakagyo-ku, Kyoto, Japan) to examine the separation efficiency. Water flux (J) of the membrane was calculated by measuring the time needed to collect some volume of permeate. This network membrane demonstrated good mechanical flexibility and robustness, which are critical for practical applications. This network membrane also showed high separation efficiency (99.9%) for oil/water emulsions with oil droplet size down to 3 µm, and meanwhile, has high water permeation flux (6.8 × 10³ L m⁻² h⁻¹ bar⁻¹) at low operation pressure. The high water flux is attributed to the interconnected scaffold-like structure throughout the whole membrane, while the high oil separation efficiency is attributed to the nanofiber-made nanoporous selective layer. Moreover, the economic materials and low-cost fabrication process of this membrane indicate its great potential for large-scale industrial applications.

Keywords: membrane, inorganic nanofibers, oil/water separation, emulsions

Procedia PDF Downloads 172

Authors: N. C. Okoronkwo, I. E. Mbaeyi-Nwaoha, C. P. Agbata

Abstract:

Childhood malnutrition is one of the most persistent public health problems throughout developing countries, including Nigeria. Demographic and Health survey data from twenty-one developing countries indicated that poor complementary feeding of children aged 6- 23 months contributes to negative growth trends. To reduce malnutrition among children in the society, formulation of complimentary food rich in essential nutrient for optimum growth and development of infants is essential. This study focused on the evaluation of complementary food produced by solid-state fermentation of Acha and Soybean using Rhizopus oligosporus (2710) and Orange-fleshed sweet potatoes (OFSP) using Lactobacillus planterum (B-41621). The raw materials were soaked separately, each in four volumes of 0.9M acetic acid for 16 hours, rinsed with clean water, steam cooked and cooled. Solid-state fermentation (SSF) was carried out by inoculating Acha and Soybean with spore suspension (1x 10⁶spores/ml) of Rhizopus oligosporus (2710) and OFSP with spore suspension (1x 106spores/ml) of Lactobacillus planterum (B-41621). Fermentation which lasted for 72hours was carried out with 24hours sampling. The samples were blended in the following ratios: Acha and soybean 100: 100 (AS), Acha/soybean and OFSP 50: 50(ASO), made into gruel and compared with a commercial infant formula (Cerelac) which served as the control (CTRL). The samples were analyzed for proximate composition using AOAC methods and sensory attributes using a hedonic scale. Results showed that moisture, crude protein, fibre and ash content increased significantly (p<0.05) as fermentation progressed, while carbohydrate and fat content decreased. The protein, moisture, fibre and ash content ranged from 17.10-19.02%, 54.97-56.27%, 7.08-7.60% and2.09-2.38%, respectively, while carbohydrate and fat content ranged from 12.95-10.21% and 5.81-4.52%, respectively. In sensory scores, there were no significant (p>0.05) difference between the average mean scores of colours, texture and consistency of the samples. The sensory score for the overall acceptability ranged from 6.20-7.80. Sample CTRL had the highest score, while sample ASO had the least score. There was no significant (p>0.05) difference between samples CTRL and AS. Solid-state fermentation improved the nutritional content and flavour of the developed complementary food, which is needed for infant growth and development.

Keywords: Complementary food, malnutrition, proximate composition, solid-state fermentation

Procedia PDF Downloads 157

Authors: Abdulnasir Hossen, Alaa Barhoum, Deepali Jaju, V. Gowri, L. Al-Kharusi, M. Hassan, K. Al-Hashmi

Abstract:

Preeclampsia is a pregnancy disorder associated with increase in blood pressure and excess amount of protein in the urine. HRV analysis has been used by many researchers to identify preeclamptic pregnancy from normal pregnancy. A study in this regard to identify preeclamptic pregnancy in Oman from normal pregnant was conducted on 40 subjects (20 patients and 20 normal). The subjects were collected from two hospitals in Oman. A Fast Fourier transform (FFT) spectral analysis has shown that patients with preeclamptic pregnancy have a reduction in the power of the HF band and an increase in the power of the LF band of HRV compared with subjects with normal pregnancy. The accuracy of identification obtained was 80%.

Keywords: preelampsia, pregnancy hypertension, normal pregnant, FFT, spectral analysis, HRV

Procedia PDF Downloads 556

Authors: D. Falliano, D. De Domenico, G. Ricciardi, E. Gugliandolo

Abstract:

This paper is focused on the mechanical characterization of foamed concrete specimens with protein-based foaming agent. Unlike classic foamed concrete, a peculiar property of the analyzed foamed concrete is the extrudability, which is achieved via a specific additive in the concrete mix that significantly improves the cohesion and viscosity of the fresh cementitious paste. A broad experimental campaign was conducted to evaluate the compressive strength and the indirect tensile strength of the specimens. The study has comprised three different cement types, two water/cement ratios, three curing conditions and three target dry densities. The variability of the strength values upon the above mentioned factors is discussed.

Keywords: cement type, curing conditions, density, extrudable concrete, foamed concrete, mechanical characterization

Procedia PDF Downloads 265

Authors: Adebola Ajayi

Abstract:

Ocimum gratissimum belongs to the Lamiaceae family and is know generally as African Basil. Ocimum gratissimum leaves are widely used as local condiments in diets. The leaves were destalked sorted, washed with potable water to remove dirts, air dried for 14 days under ambient temperature and milled into powder. The proximate composition and mineral contents of Ocimum gratissimum leaves were investigated. The proximate analysis showed the moisture, crude, protein, total ash, crude fiber, crude lipid and total carbohydrate contents were 10.72±0.01%, 12.98±0.10%, 10.95±0.42, 10.21±0.04%, 4.81±0.04% and 49.01±0.25% respectively. The results of the analysis showed that Ocimum gratissimum could be a good source of important food nutrients.

Keywords: African Basil, drying, Ocimum gratissimum, proximate

Procedia PDF Downloads 200

Authors: Mahmoodreza Tahamtan

Abstract:

Malignant surface epithelial ovarian tumors(SEOT) account for approximately 90% of primary ovarian cancer. We evaluate the immunohistochemical expression of WT1 protein among different histologic subtypes of SEOT. Immunohistochemistry for WT1 was done on 35 serous cystadenocarcinomas, 9 borderline serous tumors. A tumor was considered negative if < 1% of tumor cells were stained.Positive reactions were graded as follows:1+,1%-24%; 2+,25%-49%; 3+,50%-74%; 4+,75%-100%. Of the 35 cases of ovarian serous cystadenocarcinoma 30(85.7%)were diffusely positive(3+,4+),4 showed reactivity of < 50% of the tumor cells(1+,2+) and one were negative. All 9 borderline serous tumors showed immunoreactivity with WT1. WT1 is a good marker to distinguish primary ovarian serous carcinomas from other surface epithelial tumors.

Keywords: WT1, ovary, malignant, epithelial tumors

Procedia PDF Downloads 102

Authors: A. U. Adamu, E. D Paul, C. E. Gimba, I. G. Ndukwe

Abstract:

The phytochemical and proximate composition of Aspillia kotschyi belonging to Compositae family which is commonly used as medicinal plant in Nigeria was determined on both the Methanolic and Petroleum sprit extract of the plant. The Methanolic extract of the plant revealed the presence of carbohydrates, cardiac glyscosides, flavonoids, triterpene, and alkaloids. The Petroleum sprit extract showed the presence of only carbohydrates and alkaloid. Proximate composition analysis shows moisture content of 5.7%, total ash of 4.03%, crude protein 10.94%, fibre 9.06%, fat value 0.83%, and nitrogen free extract of 70.19%. The results of this study suggest some merit in the popular use of Aspillia kotschi in herbal medicine.

Keywords: Aspillia kotschyi, herbal medicine, phytochemical, proximate composition

Procedia PDF Downloads 366

Authors: Mads H. Clausen

Abstract:

Plant cell walls are structurally complex and contain a larger number of diverse carbohydrate polymers. These plant fibers are a highly valuable bio-resource and the focus of food, energy and health research. We are interested in studying the interplay of plant cell wall carbohydrates with proteins such as enzymes, cell surface lectins and antibodies. However, detailed molecular level investigations of such interactions are hampered by the heterogeneity and diversity of the polymers of interest. To circumvent this, we target well-defined oligosaccharides with representative structures that can be used for characterizing protein-carbohydrate binding. The presentation will highlight chemical syntheses of plant cell wall oligosaccharides from our group and provide examples from studies of their interactions with proteins.

Keywords: oligosaccharides, carbohydrate chemistry, plant cell walls, carbohydrate-acting enzymes

Procedia PDF Downloads 310

Authors: Henry O. Chibudike, Olubamike A. Adeyoju, Bolanle O. Oluwole, Kayode O. Adebowale, Bamidele I. Olu-Owolabi, Chinedum E. Chibudike

Abstract:

Studies on the Mineral Content of Solojo Flour and Protein Isolates from the two varieties (DAS and BS) of Nigeria cultivated solojo cowpeas were conducted to determine their nutritional value. These inorganic elements or minerals were classified into 3 categories: the ultra-trace minerals, which are the third category; the microelements, also known as the trace minerals, in the second category; while the first category is the macro elements, also known as major minerals. Some of the macro-elements are Ca, P, Na and Cl; the second category, micro-elements include iron, copper, cobalt, potassium, magnesium, iodine, zinc, manganese, molybdenum, F, Cr, Se and S. Results show that the proportion of Sodium (Na) which is ingested into the body in the form of NaCl through food intake maintenance of body pH and to retain water ranged from 728.97 to 253.37 ppm (72.90 to 25.34 mg/100 g); 715.24 to 235.45 ppm; 735.28 to 270.37 ppm; 726.59 to 264.35ppm, for FFDAS, FFBS, DAS and BS respectively with all values of the germinated samples all bellow the control. While FFDAS iron content ranged from 4.25 to 13.50 mg/100 g; FFBS ranged from 3.15 to 12.56 mg/100 g; DAS ranged from 3.81 to 12.90 mg/100g; BS ranged from 3.42 to 9.40 mg/100 g. The values of the germinated flours were all greater than the ungerminated flour. Iron helps to transport oxygen round the body and also helps in red blood cells building and to convert food into needed energy by the body. While Manganese an element that is needed in micro quantity but necessary to convert food into energy, is also crucial for healthy bone and cartilage creation. Results also show that zinc quantity increased as germination proceeded, and the values ranged from 38.80 ppm to 230.00 ppm (3.880 mg/100 g to 23.00 mg/100 g; 0.003880% to 0.0230%); 40.84 to 250.01 ppm; 32.85 to 93.41 ppm; 37.07 to 115.00 ppm, for FFDAS, FFBS, DAS and BS respectively. The Ca content improved significantly (p<0.05) with sprouting; the value extended from 250.56 ppm to 760.03 ppm (25.056 to 76.00 mg/100g or 0.0251 to 0.0760 %); 400.40 to 998.22 ppm; 116.87 to 195.69 ppm; 113.48 to 220.75 ppm, for FFDAS, FFBS, DAS and BS respectively. Zinc element although needed at the micro level in the body, is essential for a strong immune system to keep the body in good health. It is also crucial for the maintenance of a healthy sense of taste and odor, while Calcium is critical for strong bones and teeth, blood coagulation, and muscle tightening and relaxation. Magnesium is needed to build enzymes and antioxidants and also for healthy bones, while Potassium is needed to maintain water balance, muscle movement, and nerve impulses. It functions in conjunction with Na to regulate blood pressure.

Keywords: Solojo cowpea, underutilized legumes, protein isolates, BS, DAS, ungerminated

Procedia PDF Downloads 58

Authors: Indu Gaur, Neha Bhadauria, Shilpi Shilpi, Susmita Goswami, Prem D. Sharma, Prabir K. Paul

Abstract:

The concept of organic agriculture has been accepted as novelty in Indian society, but there is no data available on the human pathogens colonizing plant parts due to such practices. Also, the pattern and mechanism of their colonization need to be understood in order to devise possible strategies for their prevention. In the present study, human pathogenic bacteria were isolated from organically grown tomato plants and five of them were identified as Klebsiella pneumoniae, Enterobacter ludwigii, Serratia fonticola, Stenotrophomonas maltophilia and Chryseobacterium jejuense. Tomato plants were grown in controlled aseptic conditions with 25±1˚C, 70% humidity and 12 hour L/D photoperiod. Six weeks old plants were divided into 6 groups of 25 plants each and treated as follows: Group 1: K. pneumonia, Group 2: E. ludwigii, Group 3: S. fonticola, Group 4: S. maltophilia, Group 5: C. jejuense, Group 6: Sterile distilled water (control). The inoculums for all treatments were prepared by overnight growth with uniform concentration of 108 cells/ml. Leaf samples from above groups were collected at 0.5, 2, 4, 6 and 24 hours post inoculation for the colony forming unit counts (CFU/cm2 of leaf area) of individual pathogens using leaf impression method. These CFU counts were used for the in vivo colonization assay and adherence assay of individual pathogens. Also, resistance of these pathogens to at least 12 antibiotics was studied. Based on these findings S. fonticola was found to be most prominently colonizing the phylloplane of tomato and was further studied. Tomato plants grown in controlled aseptic conditions same as mentioned above were divided into 2 groups of 25 plants each and treated as follows: Group 1: S. fonticola, Group 2: Sterile distilled water (control). Leaf samples from above groups were collected at 0, 24, 48, 72 and 96 hours post inoculation and homogenized in suitable buffers for surface and cell wall protein isolation. Protein samples thus obtained were subjected to isocratic SDS-gel electrophoresis and analyzed. It was observed that presence of S. fonticola could induce the expression of at least 3 additional cell wall proteins at different time intervals. Surface proteins also showed variation in the expression pattern at different sampling intervals. Further identification of these proteins by MALDI-MS and bioinformatics tools revealed the gene(s) involved in the interaction of S. fonticola with tomato phylloplane.

Keywords: cell wall proteins, human pathogenic bacteria, phylloplane, solanum lycopersicum

Procedia PDF Downloads 228

Authors: Zohar Manber, Ran Elkon

Abstract:

Accumulation of somatic mutations (SMs) in the genome is a major driving force of cancer development. Most SMs in the tumor's genome are functionally neutral; however, some cause damage to critical processes and provide the tumor with a selective growth advantage (termed cancer driver mutations). Current research on functional significance of SMs is mainly focused on finding alterations in protein coding sequences. However, the exome comprises only 3% of the human genome, and thus, SMs in the noncoding genome significantly outnumber those that map to protein-coding regions. Although our understanding of noncoding driver SMs is very rudimentary, it is likely that disruption of regulatory elements in the genome is an important, yet largely underexplored mechanism by which somatic mutations contribute to cancer development. The expression of most human genes is controlled by multiple enhancers, and therefore, it is conceivable that regulatory SMs are distributed across different enhancers of the same target gene. Yet, to date, most statistical searches for regulatory SMs have considered each regulatory element individually, which may reduce statistical power. The first challenge in considering the cumulative activity of all the enhancers of a gene as a single unit is to map enhancers to their target promoters. Such mapping defines for each gene its set of regulating enhancers (termed "set of regulatory elements" (SRE)). Considering multiple enhancers of each gene as one unit holds great promise for enhancing the identification of driver regulatory SMs. However, the success of this approach is greatly dependent on the availability of comprehensive and accurate enhancer-promoter (E-P) maps. To date, the discovery of driver regulatory SMs has been hindered by insufficient sample sizes and statistical analyses that often considered each regulatory element separately. In this study, we analyzed more than 2,500 whole-genome sequence (WGS) samples provided by The Cancer Genome Atlas (TCGA) and The International Cancer Genome Consortium (ICGC) in order to identify such driver regulatory SMs. Our analyses took into account the combinatorial aspect of gene regulation by considering all the enhancers that control the same target gene as one unit, based on E-P maps from three genomics resources. The identification of candidate driver noncoding SMs is based on their recurrence. We searched for SREs of genes that are "hotspots" for SMs (that is, they accumulate SMs at a significantly elevated rate). To test the statistical significance of recurrence of SMs within a gene's SRE, we used both global and local background mutation rates. Using this approach, we detected - in seven different cancer types - numerous "hotspots" for SMs. To support the functional significance of these recurrent noncoding SMs, we further examined their association with the expression level of their target gene (using gene expression data provided by the ICGC and TCGA for samples that were also analyzed by WGS).

Keywords: cancer genomics, enhancers, noncoding genome, regulatory elements

Procedia PDF Downloads 104

Authors: Bello Muhammad Dogon Kade

Abstract:

The study was conducted to determine the effect of a treated groundnut (Arachis hypogaea) shell in a complete diet on blood metabolites and microbial load in fattening Yankasa rams. The study was conducted at the Teaching and Research Farm (Small Ruminants Unit of Animal Science Department, Faculty of Agriculture, Ahmadu Bello University, Zaria. Each kilogram of groundnut shell was treated with 5% urea and 5% lime for treatments 2 (UTGNS) and 3 (LTGNS), respectively. For treatment 4 (ULTGNS), 1 kg of groundnut shell was treated with 2.5% urea and 2.5% lime, but the shell in treatment 1 was not treated (UNTGNS). Sixteen Yankasa rams were used and randomly assigned to the four treatment diets with four animals per treatment in a completely randomized design (CRD). The diet was formulated to have 14% crude protein (CP) content. Rumen fluid was collected from each ram at the end of the experiment at 0 and 4 hours post-feeding. The samples were then put in a 30 ml bottle and acidified with 5 drops of concentrated sulphuric (0.1N H₂SO4) acid to trap ammonia. The results of the blood metabolites showed that the mean values of NH₃-N differed significantly (P<0.05) among the treatment groups, with rams in the ULTGNS diet having the highest significant value (31.96 mg/L). TVFs were significantly (P<0.05) higher in rams fed UNTGNS diet and higher in total nitrogen; the effect of sampling periods revealed that NH3N, TVFs and TP were significantly (P<0.05) higher in rumen fluid collected 4hrs post feeding among the rams across the treatment groups, but rumen fluid pH was significantly (p<0.05) higher in 0-hour post-feeding in all the rams in the treatment diets. In the treatment and sampling period’s interaction effects, animals on the ULTGNS diet had the highest mean values of NH3N in both 0 and 4 hours post-feeding and were significantly (P<0.5) higher compared to rams on the other treatment diets. Rams on the UTGNS diet had the highest bacteria load of 4.96X105/ml, which was significantly (P<0.05) higher than a microbial load of animals fed UNTGNS, LTGNS and ULTGNS diets. However, protozoa counts were significantly (P<0.05) higher in rams fed the UTGNS diet than those followed by the ULTGNS diet. The results showed that there was no significant difference (P>0.05) in the bacteria count of the animals at both 0 and 4 hours post-feeding. But rumen fungi and protozoa load at 0 hours were significantly (P<0.05) higher than at 4 hours post-feeding. The use of untreated ground groundnut shells in the diet of fattening Yankasa ram is therefore recommended.

Keywords: blood metabolites, microbial load, volatile fatty acid, ammonia, total protein

Procedia PDF Downloads 67

Authors: Alena O. Stepanova, Vera S. Chernonosova, Tatyana S. Godovikova, Konstantin A. Bulatov, Andrey Y. Patrushev, Pavel P. Laktionov

Abstract:

Trauma and arthrosis, not to mention cartilage destruction in overweight and elders put hyaline cartilage lesion among the most frequent diseases of locomotor system. These problems combined with low regeneration potential of the cartilage make regeneration of articular cartilage a high-priority task of tissue engineering. Many types of matrices, the procedures of their installation and autologous chondrocyte implantation protocols were offered, but certain aspects including adhesion of the implant with surrounding cartilage/bone, prevention of the ossification and fibrosis were not resolved. Simplification and acceleration of the procedures resulting in restoration of normal cartilage are also required. We have demonstrated that human chondroblasts can be successfully cultivated at the surface of electrospun scaffolds and produce extracellular matrix components in contrast to chondroblasts grown in homogeneous hydrogels. To restore cartilage we offer to use stacks of electrospun scaffolds fixed with photopolymerized solution of prepared from gelatin and chondroitin-4-sulfate both modified by glycidyl methacrylate and non-toxic photoinitator Darocur 2959. Scaffolds were prepared from nylon 6, polylactide-co-glicolide and their mixtures with modified gelatin. Illumination of chondroblasts in photopolymerized solution using 365 nm LED light had no effect on cell viability at compressive strength of the gel less than0,12 MPa. Stacks of electrospun scaffolds provide good compressive strength and have the potential for substitution with cartilage when biodegradable scaffolds are used. Vascularization can be prevented by introduction of biostable scaffolds in the layers contacting the subchondral bone. Studies of two-component materials (2-3 sheets of electrospun scaffold) implanted in the knee-joints of rabbits and fixed by photopolymerization demonstrated good crush resistance, biocompatibility and good adhesion of the implant with surrounding cartilage. Histological examination of the implants 3 month after implantation demonstrates absence of any inflammation and signs of replacement of the biodegradable scaffolds with normal cartilage. The possibility of intraoperative population of the implants with autologous cells is being investigated.

Keywords: chondroblasts, electrospun scaffolds, hyaline cartilage, photopolymerized gel

Procedia PDF Downloads 283

Authors: Farhad Aalizadeh, Ali Moosavi

Abstract:

When a blood vessel is injured, the cells of your blood bond together to form a blood clot. The blood clot helps you stop bleeding. Blood clots are made of a combination of blood cells, platelets(small sticky cells that speed up the clot-making process), and fibrin (protein that forms a thread-like mesh to trap cells). Doctors call this kind of blood clot a “thrombus.”We study the effects of different parameters on the deposition of Nanoparticles on the surface of a bump in the blood vessels by the magnetic field. The Maxwell and the flow equations are solved for this purpose. It is assumed that the blood is non-Newtonian and the number of particles has been considered enough to rely on the results statistically. Using MHD and its property it is possible to control the flow velocity, remove the fouling on the walls and return the system to its original form.

Keywords: MHD, fouling, in-vivo, blood clots, simulation

Procedia PDF Downloads 469

Authors: Peter. M. Kusen, Georg Wandrey, Christopher Probst, Dietrich Kohlheyer, Jochen Buchs, Jorg Pietruszkau

Abstract:

Light as a stimulus provides the capability to develop regulation techniques for customizable gene expression. A great advantage is the extremely flexible and accurate dosing that can be performed in a non invasive and sterile manner even for high throughput technologies. Therefore, light regulation in a multiwell microbioreactor system was realized providing the opportunity to control gene expression with outstanding complexity. A light-regulated gene expression system in Saccharomyces cerevisiae was designed applying the strategy of caged compounds. These compounds are photo-labile protected and therefore biologically inactive regulator molecules which can be reactivated by irradiation with certain light conditions. The “caging” of a repressor molecule which is consumed after deprotection was essential to create a flexible expression system. Thereby, gene expression could be temporally repressed by irradiation and subsequent release of the active repressor molecule. Afterwards, the repressor molecule is consumed by the yeast cells leading to reactivation of gene expression. A yeast strain harboring a construct with the corresponding repressible promoter in combination with a fluorescent marker protein was applied in a Photo-BioLector platform which allows individual irradiation as well as online fluorescence and growth detection. This device was used to precisely control the repression duration by adjusting the amount of released repressor via different irradiation times. With the presented screening platform the regulation of complex expression procedures was achieved by combination of several repression/derepression intervals. In particular, a stepwise increase of temporally-constant expression levels was demonstrated which could be used to study concentration dependent effects on cell functions. Also linear expression rates with variable slopes could be shown representing a possible solution for challenging protein productions, whereby excessive production rates lead to misfolding or intoxication. Finally, the very flexible regulation enabled accurate control over the expression induction, although we used a repressible promoter. Summing up, the continuous online regulation of gene expression has the potential to synchronize gene expression levels to optimize metabolic flux, artificial enzyme cascades, growth rates for co cultivations and many other applications addicted to complex expression regulation. The developed light-regulated expression platform represents an innovative screening approach to find optimization potential for production processes.

Keywords: caged-compounds, gene expression regulation, optogenetics, photo-labile protecting group

Procedia PDF Downloads 326

Authors: Saeid Doaei

Abstract:

The various studies have examined the relationship between FTO gene expression and macronutrients levels. In order to obtain better viewpoint from this interactions, all of the existing studies were reviewed systematically. All published papers have been obtained and reviewed using standard and sensitive keywords from databases such as CINAHL, Embase, PubMed, PsycInfo, and the Cochrane, from 1990 to 2016. The results indicated that all of 6 studies that met the inclusion criteria (from a total of 428 published article) found FTO gene expression changes at short-term follow-ups. Four of six studies found an increased FTO gene expression after calorie restriction, while two of them indicated decreased FTO gene expression. The effect of protein, carbohydrate and fat were separately assessed and suggested by all of six studies. In conclusion, the level of FTO gene expression in hypothalamus is related to macronutrients levels. Future research should evaluate the long-term impact of dietary interventions.

Keywords: obesity, gene expression, FTO, macronutrients

Procedia PDF Downloads 267

Authors: Nikhil Agrawal, Adam A. Skelton

Abstract:

Alzheimer’s disease (AD) is one of the most common forms of dementia, which is caused by misfolding and aggregation of amyloid beta (Aβ) peptides into amyloid-β fibrils (Aβ fibrils). To disrupt the remodeling of Aβ fibrils, a number of candidate molecules have been proposed. To study the molecular mechanisms of Aβ fibrils remodeling we performed a series of all-atom molecular dynamics simulations, a total time of 3µs, in explicit solvent. Several previously undiscovered candidate molecule-Aβ fibrils binding modes are unraveled; one of which shows the direct conformational change of the Aβ fibril by understanding the physicochemical factors responsible for binding and subsequent remodeling of Aβ fibrils by the candidate molecule, open avenues into structure-based drug design for AD can be opened.

Keywords: alzheimer’s disease, amyloid, MD simulations, misfolded protein

Procedia PDF Downloads 347

Authors: Paula K. Novelli, Margarida M. Barros, Luciana F. Flueri

Abstract:

Utilization of agricultural by-products, wheat ban and soybean bran, as substrate for solid state fermentation (SSF) was studied, aiming the achievement of different enzymes from Aspergillus sp. with distinct biological characteristics and its application and improvement on animal nutrition. Aspergillus niger and Aspergillus oryzea were studied as they showed very high yield of phytase and protease production, respectively. Phytase activity was measure using p-nitrophenilphosphate as substrate and a standard curve of p-nitrophenol, as the enzymatic activity unit was the quantity of enzyme necessary to release one μmol of p-nitrophenol. Protease activity was measure using azocasein as substrate. Activity for phytase and protease substantially increased when the different biochemical characteristics were considered in the study. Optimum pH and stability of the phytase produced by A. niger with wheat bran as substrate was between 4.0 - 5.0 and optimum temperature of activity was 37oC. Phytase fermented in soybean bran showed constant values at all pHs studied, for optimal and stability, but low production. Phytase with both substrates showed stable activity for temperatures higher than 80oC. Protease from A. niger showed very distinct behavior of optimum pH, acid for wheat bran and basic for soybean bran, respectively and optimal values of temperature and stability at 50oC. Phytase produced by A. oryzae in wheat bran had optimum pH and temperature of 9 and 37oC, respectively, but it was very unstable. On the other hand, proteases were stable at high temperatures, all pH’s studied and showed very high yield when fermented in wheat bran, however when it was fermented in soybean bran the production was very low. Subsequently the upscale production of phytase from A. niger and proteases from A. oryzae were applied as an enzyme additive in fish fed for digestibility studies. Phytases and proteases were produced with stable enzyme activity of 7,000 U.g-1 and 2,500 U.g-1, respectively. When those enzymes were applied in a plant protein based fish diet for digestibility studies, they increased protein, mineral, energy and lipids availability, showing that these new enzymes can improve animal production and performance. In conclusion, the substrate, as well as, the microorganism species can affect the biochemical character of the enzyme produced. Moreover, the production of these enzymes by SSF can be up to 90% cheaper than commercial ones produced with the same fungi species but submerged fermentation. Add to that these cheap enzymes can be easily applied as animal diet additives to improve production and performance.

Keywords: agricultural by-products, animal nutrition, enzymes production, solid state fermentation

Procedia PDF Downloads 326

Authors: Mojdeh Chelemal Dezfoul Nejad, Ali Mohammadzadeh Shobeagar, Mehrzad Mesbah

Abstract:

This study was conducted in order to characterize the different levels of dietary vitamin D on some of biochemical parameters of Cyprinus carpio. For this purpose, 180 pieces of Cyprinus carpio with an average weight of 20-25 grams were divided into four treatments and each treatment was divided into three replications and treatments were fed at three different doses (1000 IU, 3000 IU, 5000 IU) of vitamin D for 60 days. The fish were fed 3% of their wet b.wt. per day for a 60 days period. Blood samples were obtained from six fish of each tank at the end of experiment. Based on the results significant difference was observed on the mean amount of total protein, urea, glucose and cholesterol between treatments (p < 0.05). But, there was no significant difference in the mean amount of triglyceride and albumin with the different diets designed for this experiment (p > 0.05).

Keywords: Cyprinus carpio, vitamin D, biochemical parameters, glucose

Procedia PDF Downloads 349

Authors: Xi Chen, King-Yip Cheng

Abstract:

Hypercholesterolemia, characterized by high low-density lipoprotein cholesterol (LDL-C), raises cardiovascular events in patients with type 2 diabetes (T2D). Although several drugs, such as statin and PCSK9 inhibitors, are available for the treatment of hypercholesterolemia, they exert detrimental effects on glucose metabolism and hence increase the risk of T2D. On the other hand, the drugs used to treat T2D have minimal effect on improving the lipid profile. Therefore, there is an urgent need to develop treatments that can simultaneously improve glucose and lipid homeostasis. Adaptor protein phosphotyrosine interacting with PH domain and leucine zipper 2 (APPL2) causes insulin resistance in the liver and skeletal muscle via inhibiting insulin and adiponectin actions in animal models. Single-nucleotide polymorphisms in the APPL2 gene were associated with LDL-C, non-alcoholic fatty liver disease, and coronary artery disease in humans. The aim of this project is to investigate whether APPL2 antisense oligonucleotide (ASO) can alleviate dietary-induced T2D and hypercholesterolemia. High-fat diet (HFD) was used to induce obesity and insulin resistance in mice. GalNAc-conjugated APPL2 ASO (GalNAc-APPL2-ASO) was used to silence hepatic APPL2 expression in C57/BL6J mice selectively. Glucose, lipid, and energy metabolism were monitored. Immunoblotting and quantitative PCR analysis showed that GalNAc-APPL2-ASO treatment selectively reduced APPL2 expression in the liver instead of other tissues, like adipose tissues, kidneys, muscle, and heart. The glucose tolerance test and insulin sensitivity test revealed that GalNAc-APPL2-ASO improved glucose tolerance and insulin sensitivity progressively. Blood chemistry analysis revealed that the mice treated with GalNAc-APPL2-ASO had significantly lower circulating levels of total cholesterol and LDL cholesterol. However, there was no difference in circulating levels of high-density lipoprotein (HDL) cholesterol, triglyceride, and free fatty acid between the mice treated with GalNac-APPL2-ASO and GalNAc-Control-ASO. No obvious effect on food intake, body weight, and liver injury markers after GalNAc-APPL2-ASO treatment was found, supporting its tolerability and safety. We showed that selectively silencing hepatic APPL2 alleviated insulin resistance and hypercholesterolemia and improved energy metabolism in the dietary-induced obese mouse model, indicating APPL2 as a therapeutic target for metabolic diseases.

Keywords: APPL2, antisense oligonucleotide, hypercholesterolemia, type 2 diabetes

Procedia PDF Downloads 67

Authors: M. Chelemal Dezfoul Nejad, M. Moradi, M. Mesbah, M. Javaheri

Abstract:

This study was conducted in order to characterize the different levels of dietary vitamin C on some of biochemical parameters of Barbus grypus. For this purpose 300 Barbus grypus were divided into 15 groups. Five levels of vitamin C (0, 200, 400, 800, 1600 mg kg-1 diet) and their combination were used to prepare five experimental diets. The fish were fed 3% of their wet b.wt. per day for a 60 days period. Blood samples were obtained from six fish of each tank at the end of experiment. The results reveal that fish fed diets containing 1600 mg kg-1 vitamin C had a significant decrease in the mean amount of cholesterol, glucose and triglyceride (p<0.05). Also, there was no significant difference in the mean amount of total protein, albumin, BuN, phosphorus, sodium and potassium between the fish fed with the different diets designed for this experiment (p>0.05).

Keywords: Barbus grypus, vitamin C, biochemical parameters, diet

Procedia PDF Downloads 453

Authors: Muhammad Naeem Chaudhry, Fahim Nawaz, Rana Nauman Shabbir

Abstract:

New strategies aimed at increasing the resilience of crop plants to the negative effects of climate change represent important research priorities of plant scientists. The use of soil microorganisms to alleviate abiotic stresses like drought has gained particular importance in recent past. A field experiment was planned to investigate the effect of phosphorous solubilizing bacteria on yield and seed quality of Camelina (Camelina sativa L.) under water deficit conditions. The study was conducted at Agronomic Research Farm, University College of Agriculture and Environmental Sciences, The Islamia University Bahawalpur, during 4th week of November, 2013. The available seeds of Camelina sativa were inoculated with two bacterial strains (pseudomonas and Bacillus spp.) and grown under various water stress levels i.e. D0, (four irrigations), D3 (three irrigation), D2 (two irrigations), and D1 (one irrigation). The results revealed that drought stress significantly reduced the plant growth and yield, consequently reducing protein contents and oil concentration in camelina. The exposure to drought stress decreased plant height (16%), plant population (27%), number of fertile branches (41-59%), number of pods per plant (35%) and seed per pod (33%). Drought stress also exerted a negative impact on yield characteristics by reducing the 1000-seed weight (65%), final seed yield (52%), biological yield (22%) and harvest index (39%) of camelina. However, the inoculation of seeds with Pseudomonas and Bacillus spp. promoted the plant growth characterized by increased plant height and enhanced plant population. It was noted that inoculation of seeds with Pseudomonas resulted in the maximum plant population (113.4 cm), primary branches (19 plant-1), and number of pods (664 plant-1), whereas Bacillus inoculation resulted in maximum plant height (113.4 cm), seeds per pod (15.9), 1000-seed weight (1.85 g), and seed yield (3378.8 kg ha-1). Moreover, the inoculation with Bacillus also significantly improved the quality attributes of camelina and gave 3.5% and 2.1% higher oil contents than Pseudomonas and control (no-inoculation), respectively. Similarly, the same strain also resulted in maximum protein contents (33.3%). Our results confirmed the hypothesis that inoculation of seeds with phosphorous solubilizing bacterial strains is an effective, viable and environment-friendly approach to improve yield and quality of camelina under water deficit conditions. However, further studies are suggested to investigate the physiological and molecular processes, stimulated by bacterial strains, for increasing drought tolerance in food crops.

Keywords: Camelina, drought stress, phosphate solubilizing bacteria, seed quality

Procedia PDF Downloads 259

Authors: Nagia Ali, R. S. R. El-Mohamedy

Abstract:

The purpose of this paper is to irradiate the dyes biologically through the use of Moreinga oleifera. The study confirms the potential use of Moringa oleifera in decolourization of dyes and thus opens up a scope for future analysis pertaining to its performance in treatment of textile effluent. In this paper, the ability of natural products in removing dyes was tested using two reactive dyes and one acid dye. After a preliminary screening for dye removal capacity, a vegetal protein extract derived from Moeringa oleifera seed was fully studied. The influences of several parameters such as pH, temperature or initial dye concentration were tested and the behavior of coagulants was compared. It was found that dye removal decreased as pH increased. Temperature did not seem to have a considerable effect, while initial dye concentration appeared to be a very important variable.

Keywords: Moreinga oleifera, decolourization, waste water, reactive dyes, acid dyes

Procedia PDF Downloads 366

Authors: Adebola Ajayi, Olakunle M. Makanjuola

Abstract:

The effects of particle sizes of maize flour on the quality of traditional maize snack (Kokoro) were investigated. Maize flour of different sieve mesh sizes of 1.00mm, 1.9 mm, 1.4 mm, 1.68 mm and 2.0 mm was used to produce Kokoro. The samples were analysed for protein, fat, moisture content, crude fibre, ash and sensory evaluation. The various mixture obtained were separately processed into snacks following essential traditional method of production. The result of the sensory evaluation showed that Kokoro of sample 546 using 1.0mm mesh sieve size was the most preferred and sample 513 using 2.00 was least preferred. The result revealed that the more the maize was well blended the more acceptable the product is to the consumer.

Keywords: particle sizes, maize flour, quality, Kokoro

Procedia PDF Downloads 197

Authors: A. Mahshid Kakouei, B. Kumaran Suberamanin, C. Sabzali Musa Kahn, D. Mina Kakouei

Abstract:

This study aims to identify suitable conservative product for the conservation and restoration of historical Zelkova Carpinifolia wood located in humid weather. The superficial properties and hardness of 14 compounds treated with several consolidants were compared. The consolidants have been applied alone, with synthetic resin or with protein glues and natural resins by the brushing method. Colorimetric measurements, observation methods and hardness tests were conducted before and after aging to verify the possible changes of the treated wood and the consolidating resistance. The compound 1:2 of Butvar B98 and sandarac in 5% ethanol was found to be more effective, providing a suitable compound compared to the other consolidants tested.

Keywords: Zelkova carpinifolia, consolidation, synthetic resin, penetration depth, hardness

Procedia PDF Downloads 357