Search results for: protein hydrolyzate.

Authors: Y. Suranindyah, A. Astuti

Abstract:

Twelve lactating Etawah Crossedbred goats were used in this study. Goat feed consisted of Cally andra callothyrsus, Pennisetum purpureum, wheat bran and dried fermented cassava peel. The cassava peels were fermented with a traditional culture called “ragi tape" (mixed culture of Saccharomyces cerevisae, Aspergillus sp, Candida, Hasnula and Acetobacter). The goats were divided into 2 groups (Control and Treated) of six does. The experimental diet of the Control group consisted of 70% of roughage (fresh Callyandra callothyrsus and Pennisetum purpureum 60:40) and 30% of wheat bran on dry matter (DM) base. In the Treated group 30% of wheat bran was replaced with dried fermented cassava peels. Data were statistically analyzed using analysis of variance followed SPSS program. The concentration of HCN in fermented cassava peel decreased to non toxic level. Nutrient composition of dried fermented cassava peel consisted of 85.75% dry matter; 5.80% crude protein and 82.51% total digestible nutrien (TDN). Substitution of 30% of wheat bran with dried fermented cassava peel in the diet had no effect on dry matter and organic matter intake but significantly (P< 0.05) decreased crude protein and TDN consumption as well as milk yields and milk composition. The study recommended to reduced the level of substitution to less than 30% of concentrates in the diet in order to avoid low nutrient intake and milk production of goats.

Keywords: Fermented Cassava Peel, Milk Production, Composition, Etawah Crossedbred Goat.

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 3323

Authors: B. Cilek Tatar, G. Sumnu, M. Oztop, E. Ayaz

Abstract:

Encapsulation protects sensitive food ingredients against heat, oxygen, moisture and pH until they are released to the system. It can mask the unwanted taste of nutrients that are added to the foods for fortification purposes. Cherry laurels (Prunus laurocerasus) contain phenolic compounds which decrease the proneness to several chronic diseases such as types of cancer and cardiovascular diseases. The objective of this research was to study the effects of centrifugation, different coating materials and homogenization methods on microencapsulation of powders obtained from cherry laurels. In this study, maltodextrin and mixture of maltodextrin:whey protein with a ratio of 1:3 (w/w) were chosen as coating materials. Total solid content of coating materials was kept constant as 10% (w/w). Capsules were obtained from powders of freeze-dried cherry laurels through encapsulation process by silent crusher homogenizer or microfluidization. Freeze-dried cherry laurels were core materials and core to coating ratio was chosen as 1:10 by weight. To homogenize the mixture, high speed homogenizer was used at 4000 rpm for 5 min. Then, silent crusher or microfluidizer was used to complete encapsulation process. The mixtures were treated either by silent crusher for 1 min at 75000 rpm or microfluidizer at 50 MPa for 3 passes. Freeze drying for 48 hours was applied to emulsions to obtain capsules in powder form. After these steps, dry capsules were grounded manually into a fine powder. The microcapsules were analyzed for total antioxidant activity with DPPH (1,1-diphenyl-2-picrylhydrazyl) radical scavenging method. Prior to high speed homogenization, the samples were centrifuged (4000 rpm, 1 min). Centrifugation was found to have positive effect on total antioxidant activity of capsules. Microcapsules treated by microfluidizer were found to have higher total antioxidant activities than those treated by silent crusher. It was found that increasing whey protein concentration in coating material (using maltodextrin:whey protein 1:3 mixture) had positive effect on total antioxidant activity for both silent crusher and microfluidization methods. Therefore, capsules prepared by microfluidization of centrifuged mixtures can be selected as the best conditions for encapsulation of cherry laurel powder by considering their total antioxidant activity. In this study, it was shown that capsules prepared by these methods can be recommended to be incorporated into foods in order to enhance their functionality by increasing antioxidant activity.

Keywords: Antioxidant activity, cherry laurel, microencapsulation, microfluidization.

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 1309

Authors: N. Ngadi, J. Abrahamson, C. Fee, K. Morison

Abstract:

Adsorption of proteins onto a solid surface is believed to be the initial and controlling step in biofouling. A better knowledge of the fouling process can be obtained by controlling the formation of the first protein layer at a solid surface. A number of methods have been investigated to inhibit adsorption of proteins. In this study, the adsorption kinetics of

Keywords: E-casein, QCM-D, stainless steel, bimodal brush, PEG

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 1379

Authors: Marija R. Jokanović, Vladimir M. Tomović, Mihajlo T. Jović, Branislav V. Šojić, Snežana B. Škaljac, Tatjana A. Tasić, Predrag M. Ikonić

Abstract:

Proximate composition (moisture, protein, total fat, and total ash) and textural characteristics (hardness, adhesiveness, springiness, cohesiveness, chewiness and firmness and work of shear) of cooked sausages formulated from mechanically deboned chicken meat (MDCM) with addition of chicken offal (heart, gizzard or liver) were investigated. Chicken offal replaced equal weight (15 kg) of MDCM in standard sausage formulation. Regarding proximate composition sausage with heart addition was significantly (P<0.05) lower in moisture content (70.45%) than sausage with liver addition (71.35%), and significantly (P<0.05) the highest in total ash content (2.83%). Sausage with gizzard addition was significantly higher in protein content (9.77%) than sausage with liver addition (9.42%). Total fat content didn’t significantly (P>0.05) differ among all three sausages. The effect of offal addition was more notable in Warner-Bratzler shear test results than in texture profile analysis test. Firmness and work of shear were significantly different (P<0.05) among all three sausages. Sausage with liver addition was significantly (P<0.05) lower in hardness (1672 g) and chewiness (1020 g) and numerically the lowest in springiness (0.90) and adhesiveness (–70 g*s) comparing with other two sausages. Sausage with heart addition was significantly (P<0.05) higher in cohesiveness (0.74) comparing with other two sausages.

Keywords: Cooked sausage, mechanically deboned chicken meat, offal, proximate composition, texture

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 3938

Authors: Virupakshaiah DBM, Chandrakanth Kelmani, Rachanagouda Patil, Prasad Hegade

Abstract:

Severe acute respiratory syndrome (SARS) is a respiratory disease in humans which is caused by the SARS coronavirus. The treatment of coronavirus-associated SARS has been evolving and so far there is no consensus on an optimal regimen. The mainstream therapeutic interventions for SARS involve broad-spectrum antibiotics and supportive care, as well as antiviral agents and immunomodulatory therapy. The Protein- Ligand interaction plays a significant role in structural based drug designing. In the present work we have taken the receptor Angiotensin converting enzyme 2 and identified the drugs that are commonly used against SARS. They are Lopinavir, Ritonavir, Ribavirin, and Oseltamivir. The receptor Angiotensin converting enzyme 2 (ACE-2) was docked with above said drugs and the energy value obtained are as follows, Lopinavir (-292.3), Ritonavir (-325.6), Oseltamivir (- 229.1), Ribavirin (-208.8). Depending on the least energy value we have chosen the best two drugs out of the four conventional drugs. We tried to improve the binding efficiency and steric compatibility of the two drugs namely Ritonavir and Lopinavir. Several modifications were made to the probable functional groups (phenylic, ketonic groups in case of Ritonavir and carboxylic groups in case of Lopinavir respectively) which were interacting with the receptor molecule. Analogs were prepared by Marvin Sketch software and were docked using HEX docking software. Lopinavir analog 8 and Ritonavir analog 11 were detected with significant energy values and are probable lead molecule. It infers that some of the modified drugs are better than the original drugs. Further work can be carried out to improve the steric compatibility of the drug based upon the work done above for a more energy efficient binding of the drugs to the receptor.

Keywords: Protein data bank, Rasmol, Marvin sketch, Hexdocking.

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 2261

Authors: R. Razavizadeh, F. Adabavazeh, M. Rezaee Chermahini

Abstract:

Salinity is one of the most widespread agricultural problems in arid and semi-arid areas that limits the plant growth and crop productivity. In this study, the salt stress effects on protein, reducing sugar, proline contents and antioxidant enzymes activities of Carum copticum L. under in vitro conditions were studied. Seeds of C. copticum were cultured in Murashige and Skoog (MS) medium containing 0, 25, 50, 100 and 150 mM NaCl and calli were cultured in MS medium containing 1 μM 2, 4-dichlorophenoxyacetic acid, 4 μM benzyl amino purine and different levels of NaCl (0, 25, 50, 100 and 150 mM). After NaCl treatment for 28 days, the proline and reducing sugar contents of shoots, roots and calli increased significantly in relation to the severity of the salt stress. The highest amount of proline and carbohydrate were observed at 150 and 100 mM NaCl, respectively. The reducing sugar accumulation in shoots was the highest as compared to roots, whereas, proline contents did not show any significant difference in roots and shoots under salt stress. The results showed significant reduction of protein contents in seedlings and calli. Based on these results, proteins extracted from the shoots, roots and calli of C. copticum treated with 150 mM NaCl showed the lowest contents. The positive relationships were observed between activity of antioxidant enzymes and the increase in stress levels. Catalase, ascorbate peroxidase and superoxide dismutase activity increased significantly under salt concentrations in comparison to the control. These results suggest that the accumulation of proline and sugars, and activation of antioxidant enzymes play adaptive roles in the adaptation of seedlings and callus of C. copticum to saline conditions.

Keywords: Antioxidant enzymes, Carum copticum, organic solutes, salt stress.

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 1042

Authors: H. Mansoori, A. Aghazadeh, K. Nazeradl

Abstract:

The objective of this study was to investigate the effects of dietary supplementation with raw or heat-treated sunflower oil seed with two levels of 7.5% or 15% on unsaturated fatty acids in milk fat and performances of high-yielding lactating cows. Twenty early lactating Holstein cows were used in a complete randomized design. Treatments included: 1) CON, control (without sunflower oil seed). 2) LS-UT, 7.5% raw sunflower oil seed. 3) LS-HT, 7.5% heat-treated sunflower oil seed. 4) HS-UT, 15% raw sunflower oil seed. 5) HS-HT, 15% heat-treated sunflower oil seed. Experimental period lasted for 4 wk, with first 2 wk used for adaptation to the diets. Supplementation with 7.5% raw sunflower seed (LS-UT) tended to decrease milk yield, with 28.37 kg/d compared with the control (34.75 kg/d). Milk fat percentage was increased with the HS-UT treatment that obtained 3.71% compared with CON that was 3.39% and without significant different. Milk protein percent was decreased high level sunflower oil seed treatments (15%) with 3.18% whereas CON treatment is caused 3.40% protein. The cows fed added low sunflower heat-treated (LS-HT) produced milk with the highest content of total unsaturated fatty acid with 32.59 g/100g of milk fat compared with the HS-UT with 23.59 g/100g of milk fat. Content of C18 unsaturated fatty acids in milk fat increased from 21.68 g/100g of fat in the HS-UT to 22.50, 23.98, 27.39 and 30.30 g/100g of fat from the cow fed HS-HT, CON, LS-UT and LS-HT treatments, respectively. C18:2 isomers of fatty acid in milk were greater by LSHT supplementation with significant effect (P < 0.05). Total of C18 unsaturated fatty acids content was significantly higher in milk of animal fed added low heat-treated sunflower (7.5%) than those fed with high sunflower. In all, results of this study showed that diet cow's supplementation with sunflower oil seed tended to reduce milk production of lactating cows but can improve C18 UFA (Unsaturated Fatty Acid) content in milk fat. 7.5% level of sunflower oil seed that heated seemed to be the optimal source to increase UFA production.

Keywords: Fatty acid profile, milk production, sunflower seed.

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 1935

Authors: Prasad Senadheera, Younousse Saidi, Frans JM Maathuis

Abstract:

Rice seed expression (cDNA) library in the Lambda Zap 11® phage constructed from the developing grain 10-20 days after flowering was transformed into yeast for functional complementation assays in three salt sensitive yeast mutants S. cerevisiae strain CY162, G19 and Axt3K. Transformed cells of G19 and Axt3K with pYES vector with cDNA inserts showed enhance tolerance than those with empty pYes vector. Sequencing of the cDNA inserts revealed that they encode for the putative proteins with the sequence homologous to rice putative protein PROLM24 (Os06g31070), a prolamin precursor. Expression of this cDNA did not affect yeast growth in absence of salt. Axt3k and G19 strains expressing the PROLM24 were able to grow upto 400 mM and 600 mM of NaCl respectively. Similarly, Axt3k mutant with PROLM24 expression showed comparatively higher growth rate in the medium with excess LiCl (50 mM). The observation that expression of PROLM24 rescued the salt sensitive phenotypes of G19 and Axt3k indicates the existence of a regulatory system that ameliorates the effect of salt stress in the transformed yeast mutants. However, the exact function of the cDNA sequence, which shows partial sequence homology to yeast UTR1 is not clear. Although UTR1 involved in ferrous uptake and iron homeostasis in yeast cells, there is no evidence to prove its role in Na+ homeostasis in yeast cells. Absence of transmembrane regions in Os06g31070 protein indicates that salt tolerance is achieved not through the direct functional complementation of the mutant genes but through an alternative mechanism.

Keywords: Rice seed expression, salt stress, prolamin, salinitytolerance, Oryza sativa

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 1907

Authors: M. Naimi, M. B. Khaled

Abstract:

The present study consisted of an applied test in meat system to assess the effectiveness of three bio agents bacteriocinproducing strains: Lm24: Lactobacillus sakei, Lm14and Lm25: Pediococcus spp. Two tests were carried out: The ex situ test was intended for three batches added with crude bacteriocin solutions at 12.48 AU/ml for Lm25 and 8.4 AU/ml for Lm14 and Lm24. However, the in situ one consisted of four batches; three of them inoculated with one bacteriocinogenic Lm25, Lm14, Lm24, respectively. The fourth one was used in mixture: Lm14+m24 at approximately of 107 CFU/ml. The two used tests were done in the presence of the pathogen St. aureus ATCC 6538, as a test strain at 103 CFU/ml. Another batch served as a positive or a negative control was used too. The incubation was performed at 7°C. Total viable counts, staphylococci and lactic acid bacteria, at the beginning and at selected times with interval of three days were enumerated. Physico-chemical determinations (except for in situ test): pH, dry mater, sugars, fat and total protein, at the beginning and at end of the experiment, were done, according to the international norms. Our results confirmed the ex situ effectiveness. Furthermore, the batches affected negatively the total microbial load over the incubation days, and showed a significant regression in staphylococcal load at day seven, for Lm14, Lm24, and Lm25 of 0.73, 2.11, and 2.4 log units. It should be noticed that, at the last day of culture, staphylococcal load was nil for the three batches. In the in situ test, the cultures displayed less inhibitory attitude and recorded a decrease in staphylococcal load, for Lm14, Lm24, Lm25, Lm14+m24 of 0.73, 0.20, 0.86, 0.032 log units. Therefore, physicochemical analysis for Lm14, Lm24, Lm25, Lm14+m24 showed an increase in pH from 5.50 to 5.77, 6.18, 5.96, 7.22, a decrease in dry mater from 7.30% to 7.05%, 6.87%, 6.32%, 6.00%.This result reflects the decrease in fat ranging from 1.53% to 1.49%, 1.07%, 0.99%, 0.87%; and total protein from 6.18% to 5.25%, 5.56%, 5.37%, 5.5%. This study suggests that the use of selected strains as Lm25 could lead to the best results and would help in preserving and extending the shelf life of lamb meat.

Keywords: Biocontrol, in situ and ex situ, meat system, St. aureus, Lactobacillus sakei, Pediococcus spp.

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 1631

Authors: Yoshihiro Ochiai

Abstract:

Globin superfamily proteins including myoglobin and hemoglobin, have welcome new members recently, namely, cytoglobin, neuroglobin and globin X, though their physiological functions are still to be addressed. Fish are the excellent models for the study of these globins, but their characteristics have not yet been discussed to date. In the present study, attempts have been made to characterize their structural uniqueness by making use of proteomics approach. This is the first comparative study on the characterization of globin superfamily proteins from fish.

Keywords: Globin, Superfamily, Protein, Fish, Structure

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 1275

Authors: Hamada A. Ahmed, Kadry M. Sadek, Ayman E. Taha

Abstract:

The effects of basil and/or chamomile seed supplementation on the growth of Hubbard broiler chicks were evaluated. The antioxidant effects of these supplements were also assessed. 120 1-day-old broiler chicks were randomly divided into four equal groups. The control group (group 1) was fed a basal diet (BD) without supplementation. Groups 2, 3, and 4 were fed the BD supplemented with 10g basil, 10g chamomile, and 5g basil plus 5g chamomile per kg of food, respectively. Basil supplementation alone or in combination with chamomile non-significantly (P≥0.05) increased final body weight (3.2% and 0.3%, respectively) and weight gain (3.5% and 3.6%, respectively) over the experimental period. Chamomile supplementation alone non-significantly (P≥0.05) reduced final body weight and weight gain over the experimental period by 1.7% and 1.7%, respectively. In comparison to the control group, herbal seed supplementation reduced feed intake and improved the feed conversion and protein efficiency ratios. In general, basil seed supplementation stimulated chicken growth and improved the feed efficiency more effectively than chamomile seed supplementation. The antioxidant activities of basil and/or chamomile supplementation were examined in the thymus, bursa, and spleen. In chickens that received supplements, the level of malondialdehyde was significantly decreased, whereas the activities of glutathione, superoxide dismutase, and catalase were significantly increased (P<0.05). Supplementation of basil and/or chamomile did not affect blood protein levels, but had lipid-lowering effects as evidenced by reduced serum levels of total lipids, triglycerides, and cholesterol. In conclusion, supplementation of basil and/or chamomile improved growth parameters in broiler chicks and had antioxidant and blood lipid-lowering effects. These beneficial effects of basil and/or chamomile supplementation resulted in economically viable production of high-quality white meat containing no harmful residues.

Keywords: Herbal additives, basil, chamomile, broiler, growth performance, antioxidant.

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 3355

Authors: N. Umasuthan, S. D. N. K. Bathige, W. S. Thulasitha, I. Whang, J. Lee

Abstract:

The MyD88 is an evolutionarily conserved host-expressed adaptor protein that is essential for proper TLR/ IL1R immune-response signaling. A previously identified complete cDNA (1626 bp) of OfMyD88 comprised an ORF of 867 bp encoding a protein of 288 amino acids (32.9 kDa). The gDNA (3761 bp) of OfMyD88 revealed a quinquepartite genome organization composed of 5 exons (with the sizes of 310, 132, 178, 92 and 155 bp) separated by 4 introns. All the introns displayed splice signals consistent with the consensus GT/AG rule. A bipartite domain structure with two domains namely death domain (24-103) coded by 1^st exon, and TIR domain (151-288) coded by last 3 exons were identified through in silico analysis. Moreover, homology modeling of these two domains revealed a similar quaternary folding nature between human and rock bream homologs. A comprehensive comparison of vertebrate MyD88 genes showed that they possess a 5-exonic structure.In this structure, the last three exons were strongly conserved, and this suggests that a rigid structure has been maintained during vertebrate evolution.A cluster of TATA box-like sequences were found 0.25 kb upstream of cDNA starting position. In addition, putative 5'-flanking region of OfMyD88 was predicted to have TFBS implicated with TLR signaling, including copies of NFkB1, APRF/ STAT3, Sp1, IRF1 and 2 and Stat1/2. Using qPCR technique, a ubiquitous mRNA expression was detected in liver and blood. Furthermore, a significantly up-regulated transcriptional expression of OfMyD88 was detected in head kidney (12-24 h; >2-fold), spleen (6 h; 1.5-fold), liver (3 h; 1.9-fold) and intestine (24 h; ~2-fold) post-Fla challenge. These data suggest a crucial role for MyD88 in antibacterial immunity of teleosts.

Keywords: MyD88, Innate immunity, Flagellin, Genomic analysis.

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 1908

Authors: A. M. Ismaiel, Ali Hafez El-Far, Abou-Ganema I. I

Abstract:

A thirty Rahmani weaned male lambs of average body weight (27.28±1.40 kg) were randomly allotted to three similar groups, ten lambs in each, to study the benefit of commercial feed additives Tonilisat (Saccharomyces cerevisiae) and Roemin W2 (Lactobacillus acidophilus, Lactobacillus thermophilus, Bifidobacterium and Lactose) as growth promoters on lambs performance, digestibility, rumen activity and some blood constituents. The experiment lasted about 107 days. Three experimental groups were allotted as control group: received the basal ration, T1 group: received the basal ration supplemented with Tonilisat as (0.5kg/ ton concentrate feed mixture) and T2 group: received the basal ration supplemented with Roemin W2 (1kg/ ton concentrate feed mixture). Our study revealed that addition of Tonilisat significantly increased digestion coefficient of crude protein than that of the control group, Furthermore, the supplementation of Tonilisat or Roemin W2 increased (p<0.05) crude fiber digestibility than control group. Total digestible nutrients and crude digestible protein were not significantly changed between treatments. Retained nitrogen was higher in treated lamb groups than untreated but the different was non significant. Rumen activity of different rations showed that volatile fatty acids concentrations for Tonilisat and Roemin W2 groups were higher than control group, but the differences were not significant. There are no significant changes between groups in tested blood parameters but in T1 group ALT and AST were decreased. Conclusion: Supplementation of the lamb's rations with probiotics had a non significant effect (p<0.05) on blood constituents. While, growth performance and economic efficiency revealed that Tonilisat supplemented lambs had the best average daily gain followed by Roemin W2 treated group in comparison with control group. The best economic efficiency was recorded for T1 which fed Tonilisat followed by control group at whole period.

Keywords: Rahmani sheep, Tonilisat, Roemin W2, Growth, Performance.

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 2094

Authors: Arun Kumar V.A., Keshav Mohan

Abstract:

Adenylate kinase (AK) catalyse the phosphotransferase reaction plays an important role in cellular energy homeostasis. The inhibitors of bacterial AK are useful in the treatment of several bacterial infections. To the novel inhibitors of AK, docking studies performed by using the 3D structure of Bacillus stearothermophilus adenylate kinase from protein data bank (IZIP). 46 Quinoxaline analogues were docked in 1ZIP and selected the highly interacting compounds based on their binding energies, for further studies

Keywords: Kinase, quinoxaline, homeostasis, inhibitor, nucleotide.

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 1743

Authors: S. Banerjee, A. Apte Deshpande, N. Mandi, S. Padmanabhan

Abstract:

We report a novel fusion tag for expressing recombinant proteins in E. coli. The fusion tag is the C-terminus part of the human GMCSF gene comprising 45 amino acids, which aid in over expression of otherwise non expressible genes. Expression of hIFN a2b with this fusion tag also escapes the requirement of rare codons for expression. This is also a first report of a small fusion tag of human origin having affinity to heparin sepharose column facilitating the purification of fusion protein.

Keywords: fusion tag, bacterial expression, rare codons, human GMCSF

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 1898

Authors: Hanaa M. M. El-Khayat, Hoda Abdel-Hamid, Kadria M. A. Mahmoud, Hanan S. Gaber, Hoda, M. A. Abu Taleb, Hassan E. Flefel

Abstract:

Snails are considered as suitable diagnostic organisms for heavy metal–contaminated sites. Biomphalaria alexandrina snails are used in this work as pollution bioindicators after exposure to chemical mixtures consisted of heavy metals (HM); zinc (Zn), copper (Cu) and lead (Pb); and persistent organic pollutants; Decabromodiphenyl ether 98% (D) and Aroclor 1254 (A). The impacts of these tested chemicals, individual and mixtures, on liver and kidney functions, antioxidant enzymes, complete blood picture, and tissue histology were studied. Results showed that Cu was proved to be the highly toxic against snails than Zn and Pb where LC₅₀ values were 1.362, 213.198 and 277.396 ppm, respectively. Also, B. alexandrina snails exposed to the mixture of HM (¼ LC₅ Cu, Pb and Zn) showed the highest bioaccumulation of Cu and Zn in their whole tissue, the most significant increase in AST, ALT & ALP activities and the highest significant levels of total protein, albumin and globulin. Results showed significant alterations in CAT activity in snail tissue extracts while snail samples exposed to most experimental tests showed significant increase in GST activity. Snail samples that exposed to HM mixtures showed a significant decrease in total hemocytes count while snail samples that exposed to mixtures containing A & D showed a significant increase in total hemocytes and Hyalinocytes. Histopathological alterations in snail samples exposed to individual HM and their mixtures for 4 weeks showed degeneration, edema, hyper trophy and vaculation in head-foot muscle, degeneration and necrotic changes in the digestive gland and accumulation in most tested organs. Also, the hermaphrodite gland showed mature ova with irregular shape and reduction in sperm number. In conclusion, the resulted damage and alterations in B. alexandrina studied parameters can be used as bioindicators to the presence of pollutants in its habitats.

Keywords: Biomphalaria, Zn, Cu, Pb, AST, ALT, ALP, total protein albumin, globulin, CAT and Histopathology.

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 1244

Authors: Congo Tak Shing Ching, Hong-Sheng Chen, Tai-Ping Sun, Hsiu-Li Shieh

Abstract:

Nuclear matrix protein 22 (NMP22) is a FDA approved biomarker for bladder cancer. The objective of this study is to develop a simple NMP22 immumosensor (NMP22-IMS) for accurate measurement of NMP22. The NMP22-IMS was constructed with NMP22 antibody immobilized on screen-printed carbon electrodes. The construction procedures and antibody immobilization are simple. Results showed that the NMP22-IMS has an excellent (r2³0.95) response range (20 – 100 ng/mL). In conclusion, a simple and reliable NMP22-IMS was developed, capable of precisely determining urine NMP22 level.

Keywords: Bladder Cancer, Immunosensor, Impedance, NMP22

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 1657

Authors: Antonella Bandiera

Abstract:

A new sythetic gene coding for a Human Elastin-Like Polypeptide was constructed and expressed. The recombinant product was tested as coating agent to realize a surface suitable for cell growth. Coatings showed peculiar features and different human cell lines were seeded and cultured. All cell lines tested showed to adhere and proliferate on this substrate that has been shown also to exert a specific effect on cells, depending on cell type.

Keywords: elastin, recombinant protein, coating, cell adhesion.

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 1829

Authors: W. Suksombat, P. Suksombat, R. Mirattanaphrai

Abstract:

The effects of commercial or bovine yeasts on the performance and blood variables of broiler chickens intoxicated with aflatoxin were investigated in broilers. Four hundred eighty broilers (Arbor Acres; 3-wk-old) were randomly assigned to 4 groups. Each group (120 broiler chickens) was further randomly divided into 6 replicates of 20 chickens. The treatments were control diet without additives (treatment 1), 250 ppb AFB1 (treatment 2), commercial yeast, Saccharomyces cerevisiae, (CY 2.5 x 107 CFU/g) + 250 ppb AFB1 (treatment 3) and bovine yeast, Saccharomyces cerevisiae, (BY 2.5 x 107 CFU/g + 250 ppb AFB1 (treatment 4). Complete randomized design (CRD) was used in the experiment. Feed consumption and body weight were recorded at every five-day period. On day 42, carcass compositions were determined from 30 birds per treatment. While chicks were sacrificed, 3-4 ml blood sample was taken and stored frozen at (-20°C) for serum chemical analysis to determine effects of consumption of diets on blood chemistry (total protein, albumin, glucose, urea, cholesterol and triglycerides). There were no significant differences in ADFI among the treatments(P>0.05). However, BWG, FCR and mortality were highly significantly different (P<0.01) between treatments. ADG was significantly reduced (P<0.05) by aflatoxin but was unaffected by aflatoxin supplemented with either commercial or bovine yeasts (P>0.05). In terms of carcass portions, percentage of carcass was unaffected by the treatments, however, percentages of drumstick were reduced by aflatoxin and aflatoxin supplemented commercial yeast. Abdominal fat was significantly reduced (P<0.01) when commercial or bovine yeasts were added to the aflatoxin contaminated diets. Percentage of liver were significantly increased by aflatoxin contamination but were unaffected when yeasts were added to the diets. Blood chemical parameters, i.e. albumin, blood urea nitrogen and glucose were unaffected the treatments, while total protein, cholesterol and triglycerides were significantly decreased by aflatoxin. When yeasts were supplemented, such effect was not differed from the control. It is clearly indicated in the present study that supplementation of either commercial or bovine yeasts had beneficial effects on performance of broiler chickens intoxicated with aflatoxins.

Keywords: Aflatoxin, Commercial yeast, Bovine yeast, Growth performance, Blood chemical parameters, Broilers

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 2157

Authors: Paola Lecca, Lorenzo Dematte

Abstract:

Reactiondiffusion systems are mathematical models that describe how the concentration of one or more substances distributed in space changes under the influence of local chemical reactions in which the substances are converted into each other, and diffusion which causes the substances to spread out in space. The classical representation of a reaction-diffusion system is given by semi-linear parabolic partial differential equations, whose general form is ÔêétX(x, t) = DΔX(x, t), where X(x, t) is the state vector, D is the matrix of the diffusion coefficients and Δ is the Laplace operator. If the solute move in an homogeneous system in thermal equilibrium, the diffusion coefficients are constants that do not depend on the local concentration of solvent and of solutes and on local temperature of the medium. In this paper a new stochastic reaction-diffusion model in which the diffusion coefficients are function of the local concentration, viscosity and frictional forces of solvent and solute is presented. Such a model provides a more realistic description of the molecular kinetics in non-homogenoeus and highly structured media as the intra- and inter-cellular spaces. The movement of a molecule A from a region i to a region j of the space is described as a first order reaction Ai k- → Aj , where the rate constant k depends on the diffusion coefficient. Representing the diffusional motion as a chemical reaction allows to assimilate a reaction-diffusion system to a pure reaction system and to simulate it with Gillespie-inspired stochastic simulation algorithms. The stochastic time evolution of the system is given by the occurrence of diffusion events and chemical reaction events. At each time step an event (reaction or diffusion) is selected from a probability distribution of waiting times determined by the specific speed of reaction and diffusion events. Redi is the software tool, developed to implement the model of reaction-diffusion kinetics and dynamics. It is a free software, that can be downloaded from http://www.cosbi.eu. To demonstrate the validity of the new reaction-diffusion model, the simulation results of the chaperone-assisted protein folding in cytoplasm obtained with Redi are reported. This case study is redrawing the attention of the scientific community due to current interests on protein aggregation as a potential cause for neurodegenerative diseases.

Keywords: Reaction-diffusion systems, Fick's law, stochastic simulation algorithm.

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 1738

Authors: F. Alaa Ali, S. A. Sherein Abdelgayed, S. Osama. EL-Tawil, M. Adel Bakeer

Abstract:

Tartrazine is an organic azo dyes food additive widely used in foods, drugs, and cosmetics. The present study aimed to investigate the toxic effects of tartrazine on kidneys and liver biomarkers in addition to the investigation of oxidative stress and change of histopathological structure of liver and kidneys in 30 male rats. Tartrazine was orally administrated daily at dose 200 mg/ kg bw (1/ 10 LD₅₀) for sixty days. Serum and tissue samples were collected at the end of the experiment to investigate the underlying mechanism of tartrazine through assessment oxidative stress (Glutathione (GSH), Superoxide dismutase (SOD) and malondialdehyde (MDA) and biochemical markers (alanine aminotransferase (ALT), aspartate aminotransferase (AST), Total protein and Urea). Liver and kidneys tissue were collected and preserved in 10% formalin for histopathological examination. The obtained values were statistically analyzed by one way analysis of variance (ANOVA) followed by multiple comparison test. Biochemical analysis revealed that tartrazine induced significant increase in serum ALT, AST, total protein, urea level compared to control group. Tartrazine showed significant decrease in liver GSH and SOD where their values when compared to control group. Tartrazine induced increase in liver MDA compared to control group. Histopathology of the liver showed diffuse vacuolar degeneration in hepatic parenchyma, the portal area showed sever changes sever in hepatoportal blood vessels and in the bile ducts. The kidneys showed degenerated tubules at the cortex together with mononuclear leucocytes inflammatory cells infiltration. There is perivascular edema with inflammatory cell infiltration surrounding the congested and hyalinized vascular wall of blood vessel. The present study indicates that the subchronic effects of tartrazine have a toxic effect on the liver and kidneys together with induction of oxidative stress by formation of free radicals. Therefore, people should avoid the hazards of consuming tartrazine.

Keywords: Albino rats, tartrazine, toxicity, pathology

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 2201

Authors: Sérgio F. Sousa, Nuno M. F. S. A. Cerqueira, Marta A. S. Perez, Irina S. Moreira, António J. M.Ribeiro, Ana R. A. P. Neves, Maria J. Ramos, Pedro A. Fernandes

Abstract:

In this work we report the recent progresses that have been achieved by our group in the last half decade on the field of computational proteomics. Specifically, we discuss the application of Molecular Dynamics Simulations and Electronic Structure Calculations in drug design, in the clarification of the structural and dynamic properties of proteins and enzymes and in the understanding of the catalytic and inhibition mechanism of cancer-related enzymes. A set of examples illustrate the concepts and help to introduce the reader into this important and fast moving field.

Keywords: Enzyme, Molecular Dynamics, Protein, Quantum Mechanics.

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 1956

Authors: Ireneusz Majsterek, Anna Merecz, Agnieszka Sliwinska, Marcin Kosmalski, Jacek Kasznicki, Jozef Drzewoski

Abstract:

Oxidative stress is considered to be the cause for onset and the progression of type 2 diabetes mellitus (T2DM) and complications including neuropathy. It is a deleterious process that can be an important mediator of damage to cell structures: protein, lipids and DNA. Data suggest that in patients with diabetes and diabetic neuropathy DNA repair is impaired, which prevents effective removal of lesions. Objective: The aim of our study was to evaluate the association of the hOGG1 (326 Ser/Cys) and XRCC1 (194 Arg/Trp, 399 Arg/Gln) gene polymorphisms whose protein is involved in the BER pathway with DNA repair efficiency in patients with diabetes type 2 and diabetic neuropathy compared to the healthy subjects. Genotypes were determined by PCR-RFLP analysis in 385 subjects, including 117 with type 2 diabetes, 56 with diabetic neuropathy and 212 with normal glucose metabolism. The polymorphisms studied include codon 326 of hOGG1 and 194, 399 of XRCC1 in the base excision repair (BER) genes. Comet assay was carried out using peripheral blood lymphocytes from the patients and controls. This test enabled the evaluation of DNA damage in cells exposed to hydrogen peroxide alone and in the combination with the endonuclease III (Nth). The results of the analysis of polymorphism were statistically examination by calculating the odds ratio (OR) and their 95% confidence intervals (95% CI) using the ¤ç2-tests. Our data indicate that patients with diabetes mellitus type 2 (including those with neuropathy) had higher frequencies of the XRCC1 399Arg/Gln polymorphism in homozygote (GG) (OR: 1.85 [95% CI: 1.07-3.22], P=0.3) and also increased frequency of 399Gln (G) allele (OR: 1.38 [95% CI: 1.03-1.83], P=0.3). No relation to other polymorphisms with increased risk of diabetes or diabetic neuropathy. In T2DM patients complicated by neuropathy, there was less efficient repair of oxidative DNA damage induced by hydrogen peroxide in both the presence and absence of the Nth enzyme. The results of our study suggest that the XRCC1 399 Arg/Gln polymorphism is a significant risk factor of T2DM in Polish population. Obtained data suggest a decreased efficiency of DNA repair in cells from patients with diabetes and neuropathy may be associated with oxidative stress. Additionally, patients with neuropathy are characterized by even greater sensitivity to oxidative damage than patients with diabetes, which suggests participation of free radicals in the pathogenesis of neuropathy.

Keywords: Diabetic neuropathy, oxidative stress, gene polymorphisms, oxidative DNA damage.

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 2068

Authors: Marzhan Baimurzayeva, Alibek Utyanov, Gulnar Shabdarbaeva, Damir Khussainov

Abstract:

This article introduces the actual problem that is а study of proposed by the authors Hydrocele ointment in amino acids’ metabolism of cows’ blood in inflammation of traumatic origin. Hydrocele ointment has shown a positive effect on inflammatory process and amino acids’ metabolism of animals treated with the drug. Amino acid levels reached physiological parameters on the 10th day after treatment; in the control group this parameter was higher than normal.

Keywords: Amino acids, blood protein, Hydrocele ointment, inflammation, repair.

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 1824

Authors: T Rusu, P I Moraru, I Bogdan, A I Pop, M L Sopterean

Abstract:

The paper presents the influence of the conventional ploughing tillage technology in comparison with the minimum tillage, upon the soil properties, weed control and yield in the case of maize (Zea mays L.), soya-bean (Glycine hispida L.) and winter wheat (Triticum aestivum L.) in a three years crop rotation. A research has been conducted at the University of Agricultural Sciences and Veterinary Medicine Cluj-Napoca, Romania. The use of minimum soil tillage systems within a three years rotation: maize, soya-bean, wheat favorites the rise of the aggregates hydro stability with 5.6-7.5% on a 0-20 cm depth and 5-11% on 20-30 cm depth. The minimum soil tillage systems – paraplow, chisel or rotary grape – are polyvalent alternatives for basic preparation, germination bed preparation and sowing, for fields and crops with moderate loose requirements being optimized technologies for: soil natural fertility activation and rationalization, reduction of erosion, increasing the accumulation capacity for water and realization of sowing in the optimal period. The soil tillage system influences the productivity elements of cultivated species and finally the productions thus obtained. Thus, related to conventional working system, the productions registered in minimum tillage working represented 89- 97% in maize, 103-112% in soya-bean, 93-99% in winter-wheat. The results of investigations showed that the yield is a conclusion soil tillage systems influence on soil properties, plant density assurance and on weed control. Under minimum tillage systems in the case of winter weat as an option for replacing classic ploughing, the best results in terms of quality indices were obtained from version worked with paraplow, followed by rotary harrow and chisel. At variants worked with paraplow were obtained quality indices close to those of the variant worked with plow, and protein and gluten content was even higher. At Ariesan variety, highest protein content, 12.50% and gluten, 28.6% was obtained for the variant paraplow.

Keywords: Minimum tillage, soil properties, yields quality.

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 1919

Authors: F. Yaghobian, R. Stosch, B. Güttler

Abstract:

This study aims to demonstrate the quantification of peptides based on isotope dilution surface enhanced Raman scattering (IDSERS). SERS spectra of phenylalanine (Phe), leucine (Leu) and two peptide sequences TGQIFK (T13) and YSFLQNPQTSLCFSESIPTPSNR (T6) as part of the 22-kDa human growth hormone (hGH) were obtained on Ag-nanoparticle covered substrates. On the basis of the dominant Phe and Leu vibrational modes, precise partial least squares (PLS) prediction models were built enabling the determination of unknown T13 and T6 concentrations. Detection of hGH in its physiological concentration in order to investigate the possibility of protein quantification has been achieved.

Keywords: Surface Enhanced Raman Scattering, Quantification, Peptides.

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 1672

Authors: S. Al-Khalasi, O. Mahgoub, H. Yaakub

Abstract:

This study evaluated the use of raw or processed Prosopis juliflora (Meskit) pods as a major ingredient in a formulated ration to provide an alternative non-conventional concentrate for livestock feeding in Oman. Dry Meskit pods were reduced to lengths of 0.5- 1.0 cm to ensure thorough mixing into three diets. Meskit pods were subjected to two types of treatments; roasting and soaking. They were roasted at 150оC for 30 minutes using a locally-made roasting device (40 kg barrel container rotated by electric motor and heated by flame gas cooker). Chopped pods were soaked in tap water for 24 hours and dried for 2 days under the sun with frequent turning. The Meskit-pod-based diets (MPBD) were formulated and pelleted from 500 g/kg ground Meskit pods, 240 g/kg wheat bran, 200 g/kg barley grain, 50 g/kg local dried sardines and 10 g/kg of salt. Twenty four 10 months-old intact Omani male lambs with average body weight of 27.3 kg (± 0.5 kg) were used in a feeding trial for 84 days. They were divided (on body weight basis) and allocated to four diet combination groups. These were: Rhodes grass hay (RGH) plus a general ruminant concentrate (GRC); RGH plus raw Meskit pods (RMP) based concentrate; RGH plus roasted Meskit pods (ROMP) based concentrate; RGH plus soaked Meskit pods (SMP) based concentrate Daily feed intakes and bi-weekly body weights were recorded. MPBD had higher contents of crude protein (CP), acid detergent fibre (ADF) and neutral detergent fibre (NDF) than the GRC. Animals fed various types of MPBD did not show signs of ill health. There was a significant effect of feeding ROMP on the performance of Omani sheep compared to RMP and SMP. The ROMP fed animals had similar performance to those fed the GRC in terms of feed intake, body weight gain and feed conversion ratio (FCR).This study indicated that roasted Meskit pods based diet may be used instead of the commercial concentrate for feeding Omani sheep without adverse effects on performance. It offers a cheap alternative source of protein and energy for feeding Omani sheep. Also, it might help in solving the spread impact of Meskit trees, maintain the ecosystem and helping in preserving the local tree species.

Keywords: Growth, Meskit, Omani sheep, Prosopis juliflora.

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 2776

Authors: Mustafa M. Donma, Orkide Donma

Abstract:

Zinc (Zn) is a vital element required for growth and development particularly in children. It exhibits some protective effects against cardiovascular diseases (CVDs). Zn may be a potential biomarker of cardiovascular health. High sensitive cardiac troponin T (hs-cTnT) and cardiac myosin binding protein C (cMyBP-C) are new generation markers used for prediagnosis, diagnosis and prognosis of CVDs. The aim of this study is to determine Zn as well as new generation cardiac markers’ profiles in children with normal body mass index (N-BMI), obese (OB), morbid obese (MO) children and children with metabolic syndrome (MetS) findings. The association among them will also be investigated. Four study groups were constituted. The study protocol was approved by the institutional Ethics Committee of Tekirdag Namik Kemal University. Parents of the participants filled informed consent forms to participate in the study. Group 1 is composed of 44 children with N-BMI. Group 2 and Group 3 comprised 43 OB and 45 MO children, respectively. 45 MO children with MetS findings were included in Group 4. World Health Organization age- and sex-adjusted BMI percentile tables were used to constitute groups. These values were 15-85, 95-99 and above 99 for N-BMI, OB and MO, respectively. Criteria for MetS findings were determined. Routine biochemical analyses including Zn were performed. hs-cTnT and cMyBP-C concentrations were measured by enzyme-linked immunosorbent assay. Data were analyzed by using SPSS software. p < 0.05 was accepted as significant. Four groups were matched for age and gender. Decreased Zn concentrations were measured in Groups 2, 3 and 4 compared to Group 1. Groups did not differ from one another in terms of hs-cTnT. There were statistically significant differences between cMyBP-C levels of MetS group and N-BMI as well as OB groups. There was an increasing trend going from N-BMI group to MetS group. There were statistically significant negative correlations between Zn and hs-cTnT as well as cMyBP-C concentrations in MetS group. In conclusion, inverse correlations detected between Zn and new generation cardiac markers (hs-TnT and cMyBP-C) have pointed out that decreased levels of Zn accompany increased levels of hs-cTnT as well as cMyBP-C in children with MetS. This finding emphasizes that both Zn and these new generation cardiac markers may be evaluated as biomarkers of cardiovascular health during severe childhood obesity precipitated with MetS findings and also suggested as the messengers of the future risk in the adulthood periods of children with MetS.

Keywords: Cardiac myosin binding protein-C, cardiovascular diseases, children, high sensitive cardiac troponin T, obesity.

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 526

Authors: Shaiemaa, A. Momen, Dalia, A.M. Salem, Emad, M.S. Barakat, Mohamed, S. Salama

Abstract:

The present study has been conducted to characterize the prophenoloxidase (PPO) system of the desert locust, Schistocerca gregaria following injection of Bacillus thuringiensis kurstaki (Bt). The bulk of PPO system was associated with haemocytes and a little amount was found in plasma. This system was activated by different activators such as laminarin, lipopolysaccharide (LPS) and trypsin suggesting that the stimulatory mechanism may involve an enzyme cascade of one or more associated molecules. These activators did not activate all the molecules of the cascade. Presence of phenoloxidase activity (PO) coincides with the appearance of protein band with molecular weight (MW) 70.154 KD (Kilo Dalton).

Keywords: Schistocerca gregaria, haemolymph, proteins, prophenoloxidase system, phenoloxidase

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 1722

Authors: Chlopicka Joanna, Barton Henryk, Kryczyk Jadwiga, Francik Renata

Abstract:

Recent studies demonstrated that high-fat diet increases oxidative stress in plasma and in a variety of tissues. Many researchers have been looking for natural products, which can reverse the effect of high fat diet. Recently, buckwheat is becoming common ingredient in functional food because of it properties. In study on buckwheat, it is known that, this plant plays roles as anti-oxidative, anti-inflammatory and anti-hypertensive. Nevertheless still little is known about buckwheat groats. The aim of this study was to investigate the effects of addition of buckwheat groats to the fat diet (30% lard), on some antioxidant and oxidant stress parameters in plasma and selected tissues in Wistar rats. The experiment was carried out with three months old male Wistar rats ca. 250g of body weight fed for 5 weeks with either a high-fat (30% of lard) diet or control diet, with or without addition of buckwheat groats. In plasma biochemistry and the activities of the antioxidant enzymes were measured selected tissues: glutathione peroxidase (GPX), catalase (CAT) and the levels of total and reduced glutathione (GSH), free thiol groups (pSH), antioxidant potential of plasma (FRAP) and oxidant stress indices - proteins carbonyl groups (CO) and malonyldialdehyde concentration (MDA). Activity of catalase (CAT) in plasma of rats was significantly increased in buckwheat groats groups and activity of GPx3 in plasma of rats was decreased in buckwheat groups as compared to control group. The reduced glutathione (GSH) in plasma of rats was significantly increased and protein CO was significantly decreased in buckwheat groups as compared to controls. The lowered concentration of GSH was found in serum of rats fed buckwheat groats addition but it accompanied in 7-fold increase in reduced-to-oxidized glutatione ratio, significant increase in HDL and decrease in nonHDL concentration. Conclusions: Buckwheat groats indicate a beneficial effect in inhibiting protein and lipid peroxidation in rats and improved lipid profile. These results suggest that buckwheat groats exert a significant antioxidant potential and may be used as normal food constituent to ameliorate the oxidant-induced damage in organism. 

Keywords: Antioxidant, buckwheat, high-fat diet, rats.

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 2471