Search results for: bovine serum albumin (BSA)
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 168

Search results for: bovine serum albumin (BSA)

168 Adsorption of Bovine Serum Albumin on CeO2

Authors: Roman Marsalek

Abstract:

Preparation of nanoparticles of cerium oxide and adsorption of bovine serum albumin on them were studied. Particle size distribution and influence of pH on zeta potential of prepared CeO2 were determined. Average size of prepared cerium oxide nanoparticles was 9 nm. The simultaneous measurements of the bovine serum albumin adsorption and zeta potential determination of the (adsorption) suspensions were carried out. The adsorption isotherms were found to be of typical Langmuir type; values of the bovine serum albumin adsorption capacities were calculated. Increasing of pH led to decrease of zeta potential and decrease of adsorption capacity of cerium oxide nanoparticles. The maximum adsorption capacity was found for strongly acid suspension (am = 118 mg/g). The samples of nanoceria with positive zeta potential adsorbed more bovine serum albumin on the other hand, the samples with negative zeta potential showed little or no protein adsorption. Surface charge or better say zeta potential of CeO2 nanoparticles plays the key role in adsorption of proteins on such type of materials.

Keywords: Adsorption, BSA, cerium oxide nanoparticles, zeta potential.

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167 The Influence of Surface Potential on the Kinetics of Bovine Serum Albumin Adsorption on a Biomedical Grade 316LVM Stainless Steel Surface

Authors: Khawtar Hasan Ahmed, Sasha Omanovic

Abstract:

Polarization modulation infrared reflection absorption spectroscopy (PM-IRRAS) in combination with electrochemistry, was employed to study the influence of surface charge (potential) on the kinetics of bovine serum albumin (BSA) adsorption on a biomedical-grade 316LVM stainless steel surface is discussed. The BSA adsorption kinetics was found to greatly depend on the surface potential. With an increase in surface potential towards more negative values, both the BSA initial adsorption rate and the equilibrium (saturated) surface concentration also increased. Both effects were explained on the basis of replacement of well-ordered water molecules at the 316LVM / solution interface, i.e. by the increase in entropy of the system.

Keywords: adsorption, biomedical grade stainless steel, bovine serum albumin (BSA), electrode surface potential / charge, kinetics, PM-IRRAS, protein/surface interactions

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166 Inulinase Immobilization on Functionalized Magnetic Nanoparticles Prepared with Soy Protein Isolate Conjugated Bovine Serum Albumin for High Fructose Syrup Production

Authors: Homa Torabizadeh, Mohaddeseh Mikani

Abstract:

Inulinase from Aspergillus niger was covalently immobilized on magnetic nanoparticles (MNPs/Fe3O4) covered with soy protein isolate (SPI/Fe3O4) functionalized by bovine serum albumin (BSA) nanoparticles. MNPs are promising enzyme carriers because they separate easily under external magnetic fields and have enhanced immobilized enzyme reusability. As MNPs aggregate simply, surface coating strategy was employed. SPI functionalized by BSA was a suitable candidate for nanomagnetite coating due to its superior biocompatibility and hydrophilicity. Fe3O4@SPI-BSA nanoparticles were synthesized as a novel carrier with narrow particle size distribution. Step by step fabrication monitoring of Fe3O4@SPI-BSA nanoparticles was performed using field emission scanning electron microscopy and dynamic light scattering. The results illustrated that nanomagnetite with the spherical morphology was well monodispersed with the diameter of about 35 nm. The average size of the SPI-BSA nanoparticles was 80 to 90 nm, and their zeta potential was around −34 mV. Finally, the mean diameter of fabricated Fe3O4@SPI-BSA NPs was less than 120 nm. Inulinase enzyme from Aspergillus niger was covalently immobilized through gluteraldehyde on Fe3O4@SPI-BSA nanoparticles successfully. Fourier transform infrared spectra and field emission scanning electron microscopy images provided sufficient proof for the enzyme immobilization on the nanoparticles with 80% enzyme loading.

Keywords: High fructose syrup, inulinase immobilization, functionalized magnetic nanoparticles, soy protein isolate.

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165 Effect of Commercial or Bovine Yeasts on the Performance and Blood Variables of Broiler Chickens Intoxicated with Aflatoxins

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

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164 Protein Delivery from Polymeric Nanoparticles

Authors: G. Spada, E. Gavini, P. Giunchedi

Abstract:

Aim of this work was to compare the efficacy of two loading methods of proteins onto polymeric nanocarriers: adsorption and encapsulation methods. Preliminary studies of protein loading were done using Bovine Serum Albumin (BSA) as model protein. Nanocarriers were prepared starting from polylactic co-glycolic acid (PLGA) polymer; production methods used are two different variants of emulsion evaporation method. Nanoparticles obtained were analyzed in terms of dimensions by Dynamic Light Scattering and Loading Efficiency of BSA by Bradford Assay. Loaded nanoparticles were then submitted to in-vitro protein dissolution test in order to study the effect of the delivery system on the release rate of the protein.

Keywords: Drug delivery, nanoparticles, PLGA, proteinadsorption, protein encapsulation.

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163 Secondary Ion Mass Spectrometry of Proteins

Authors: Santanu Ray, Alexander G. Shard

Abstract:

The adsorption of bovine serum albumin (BSA), immunoglobulin G (IgG) and fibrinogen (Fgn) on fluorinated selfassembled monolayers have been studied using time of flight secondary ion mass spectrometry (ToF-SIMS) and Spectroscopic Ellipsometry (SE). The objective of the work has to establish the utility of ToF-SIMS for the determination of the amount of protein adsorbed on the surface. Quantification of surface adsorbed proteins was carried out using SE and a good correlation between ToF-SIMS results and SE was achieved. The surface distribution of proteins were also analysed using Atomic Force Microscopy (AFM). We show that the surface distribution of proteins strongly affect the ToFSIMS results.

Keywords: ToF-SIMS, Spectroscopic Ellipsometry, Protein, Atomic Force Microscopy.

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162 Comparison of Real-Time PCR and FTIR with Chemometrics Technique in Analysing Halal Supplement Capsules

Authors: Mohd Sukri Hassan, Ahlam Inayatullah Badrul Munir, M. Husaini A. Rahman

Abstract:

Halal authentication and verification in supplement capsules are highly required as the gelatine available in the market can be from halal or non-halal sources. It is an obligation for Muslim to consume and use the halal consumer goods. At present, real-time polymerase chain reaction (RT-PCR) is the most common technique being used for the detection of porcine and bovine DNA in gelatine due to high sensitivity of the technique and higher stability of DNA compared to protein. In this study, twenty samples of supplements capsules from different products with different Halal logos were analyzed for porcine and bovine DNA using RT-PCR. Standard bovine and porcine gelatine from eurofins at a range of concentration from 10-1 to 10-5 ng/µl were used to determine the linearity range, limit of detection and specificity on RT-PCR (SYBR Green method). RT-PCR detected porcine (two samples), bovine (four samples) and mixture of porcine and bovine (six samples). The samples were also tested using FT-IR technique where normalized peak of IR spectra were pre-processed using Savitsky Golay method before Principal Components Analysis (PCA) was performed on the database. Scores plot of PCA shows three clusters of samples; bovine, porcine and mixture (bovine and porcine). The RT-PCR and FT-IR with chemometrics technique were found to give same results for porcine gelatine samples which can be used for Halal authentication.

Keywords: Halal, real-time PCR, gelatin, FTIR and chemometrics.

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161 Microneedles-Mediated Transdermal Delivery

Authors: M. Petchsangsai, N. Wonglertnirant, T. Rojanarata, P. Opanasopit, T. Ngawhirunpat

Abstract:

The objective of the present study was to evaluate the potential of hollow microneedles for enhancing the transdermal delivery of Bovine Serum Albumin (MW~66,000 Da)-Fluorescein Isothiocyanate (BSA-FITC) conjugate, a hydrophilic large molecular compound. Moreover, the effect of different formulations was evaluated. The series of binary mixtures composed of propylene glycol (PG) and pH 7.4 phosphate buffer solution (PBS) was prepared and used as a medium for BSA-FITC. The results showed that there was no permeation of BSA-FITC solution across the neonatal porcine skin without using hollow microneedles, whereas the cumulative amount of BSA-FITC released at 8 h through the neonatal porcine skin was about 60-70% when using hollow microneedles. Furthermore, the results demonstrated that the higher volume of PG in binary mixtures injected, the lower cumulative amount of BSA-FITC released and release rate of BSA-FITC from skin. These release profiles of BSA-FITC in binary mixtures were expressed by Fick-s law of diffusion. These results suggest the utilization of hollow microneedle to enhance transdermal delivery of protein and provide useful information for designing an effective hollow microneedle system.

Keywords: Hydrophilic macromolecules, Microneedles, Propylene glycol, Transdermal drug delivery

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160 Application of Whole Genome Amplification Technique for Genotype Analysis of Bovine Embryos

Authors: S. Moghaddaszadeh-Ahrabi, S. Farajnia, Gh. Rahimi-Mianji, A. Nejati-Javaremi

Abstract:

In recent years, there has been an increasing interest toward the use of bovine genotyped embryos for commercial embryo transfer programs. Biopsy of a few cells in morulla stage is essential for preimplantation genetic diagnosis (PGD). Low amount of DNA have limited performing the several molecular analyses within PGD analyses. Whole genome amplification (WGA) promises to eliminate this problem. We evaluated the possibility and performance of an improved primer extension preamplification (I-PEP) method with a range of starting bovine genomic DNA from 1-8 cells into the WGA reaction. We optimized a short and simple I-PEP (ssI-PEP) procedure (~3h). This optimized WGA method was assessed by 6 loci specific polymerase chain reactions (PCRs), included restriction fragments length polymorphism (RFLP). Optimized WGA procedure possesses enough sensitivity for molecular genetic analyses through the few input cells. This is a new era for generating characterized bovine embryos in preimplantation stage.

Keywords: Whole genome amplification (WGA), Genotyping, Bovine, Preimplantation genetic diagnosis (PGD)

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159 Evaluation of Disease Risk Variables in the Control of Bovine Tuberculosis

Authors: Berrin Şentürk

Abstract:

In this study, due to the recurrence of bovine tuberculosis, in the same areas, the risk factors for the disease were determined and evaluated at the local level. This study was carried out in 32 farms where the disease was detected in the district and center of Samsun province in 2014. Predetermined risk factors, such as farm, environmental and economic risks, were investigated with the survey method. It was predetermined that risks in the three groups are similar to the risk variables of the disease on the global scale. These risk factors that increase the susceptibility of the infection must be understood by the herd owners. The risk-based contagious disease management system approach should be applied for bovine tuberculosis by farmers, animal health professionals and public and private sector decision makers.

Keywords: Bovine tuberculosis, disease management, control, outbreak, risk analysis.

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158 Enhancing Protein Incorporation in Calcium Phosphate Coating on Titanium by Rapid Biomimetic Co-Precipitation Technique

Authors: J. Suwanprateeb, F. Thammarakcharoen

Abstract:

Calcium phosphate coating (CaP) has been employed for protein delivery, but the typical direct protein adsorption on the coating led to low incorporation content and fast release of the protein from the coating. By using bovine serum albumin (BSA) as a model protein, rapid biomimetic co-precipitation between calcium phosphate and BSA was employed to control the distribution of BSA within calcium phosphate coating during biomimetic formation on titanium surface for only 6 h at 50oC in an accelerated calcium phosphate solution. As a result, the amount of BSA incorporation and release duration could be increased by using a rapid biomimetic coprecipitation technique. Up to 43 fold increases in the BSA incorporation content and the increase from 6 h to more than 360 h in release duration compared to typical direct adsorption technique were observed depending on the initial BSA concentration used during coprecipitation (1, 10 and 100 μg.ml-1). From x-ray diffraction and Fourier transform infrared spectroscopy studies, the coating composition was not altered with the incorporation of BSA by this rapid biomimetic co-precipitation and mainly comprised octacalcium phosphate and hydroxyapatite. However, the microstructure of calcium phosphate crystals changed from straight, plate-like units to curved, plate-like units with increasing BSA content.

Keywords: Biomimetic, Calcium Phosphate Coating, Protein, Titanium.

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157 Growth Performance and Blood Characteristics of Broilers Chicken Fed on Diet Containing Brewer Spent Grain at Finisher Phase

Authors: O. A. Anjola, M. A. Adejobi, L. A Tijani

Abstract:

This study was conducted to investigate the effects of brewer spent grain (BSG) on growth performance and serum biochemistry characteristics of blood of broilers chickens. Three hundred and fifteen (4 weeks old) Oba – Marshall Broilers were used for the experiment. Five experimental diets were formulated with diet 1 (T1) containing 100% soya bean meal as the control, Diet 2, 3, 4 and 5 had BSG as replacement for soya bean meal at 0%, 36%, 57%, 76% and 100% respectively. The birds were allocated into each dietary group in a completely randomized design with 63 chicks in 3 replicates of 21 chicks each. The birds were offered these diets ad libitum from four weeks old to nine weeks old (35 days). Feed intake, body weight, weight gain, and feed conversion ratio (FCR) were assessed. Blood samples were also collected to examine the effect of BSG waste on hematology and serum biochemistry of broilers. Result indicated that BSG did not significantly (P>0.05) affect feed intake and weight gain. However, FCR and final weight of finishing broilers differs significantly (P<0.05) among treatments. The blood hematology and serum biochemistry indices did not follow a particular trend. Cholesterol concentration reduced with increasing level of BSG in the diet. Hb, RBC, WBC, neutrophils, lymphocytes, heterophiles and MCHC were significant (P<0.05) while MHC and MVC were not significantly (P>0.05) affected by BSG in diets. serum total protein, albumin, and cholesterol concentration also showed significance (P<0.05) difference. Thus, BSG can replace soya bean meal up to 14% in the broiler finisher diet without deleterious effect on the growth, hematology and the serum biochemistry of broiler chicken.

Keywords: Broilers, growth performance, hematology, serum biochemistry.

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156 Nutritional Potential and Functionality of Whey Powder Influenced by Different Processing Temperature and Storage

Authors: Zarmina Gillani, Nuzhat Huma, Aysha Sameen, Mulazim Hussain Bukhari

Abstract:

Whey is an excellent food ingredient owing to its high nutritive value and its functional properties. However, composition of whey varies depending on composition of milk, processing conditions, processing method, and its whey protein content. The aim of this study was to prepare a whey powder from raw whey and to determine the influence of different processing temperatures (160 and 180 °C) on the physicochemical, functional properties during storage of 180 days and on whey protein denaturation. Results have shown that temperature significantly (P < 0.05) affects the pH, acidity, non-protein nitrogen (NPN), protein total soluble solids, fat and lactose contents. Significantly (p < 0.05) higher foaming capacity (FC), foam stability (FS), whey protein nitrogen index (WPNI), and a lower turbidity and solubility index (SI) were observed in whey powder processed at 160 °C compared to whey powder processed at 180 °C. During storage of 180 days, slow but progressive changes were noticed on the physicochemical and functional properties of whey powder. Reverse phase-HPLC analysis revealed a significant (P < 0.05) effect of temperature on whey protein contents. Denaturation of β-Lactoglobulin is followed by α-lacalbumin, casein glycomacropeptide (CMP/GMP), and bovine serum albumin (BSA).

Keywords: Whey powder, temperature, denaturation, reverse phase – HPLC.

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155 Effect of Oral Administration of “Gadagi“ Tea on Liver Function in Rats

Authors: A. M. Gadanya, M. S. Sule, M. K. Atiku

Abstract:

Effect of oral administration of “Gadagi" tea on liver function was assessed on 50 healthy male albino rats which were grouped and administered with different doses(mg/kg) i.e low dose (380mg/kg, 415mg/kg, 365mg/kg, 315mg/kg for “sak", “sada" and “magani" respectively), standard dose ( 760mg/kg, 830mg/kg, 730mg/kg for “sak-, “sada" and “magani" respectively) and high dose (1500mg/kg, 1700mg/kg and 1460mg/kg for “sak--,"sada" and “magani" groups respectively) for a period of four weeks. Animals that were not administered with the tea constituted the control group. At the end of fourth week, the animals were sacrificed and their serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), total protein (TP), albumin (ALB), and globulins (GLO) were determined. Mean serum ALT and ALP activities were significantly higher (P<0.05) in rats orally administered with high dose of “sak" and those administered with standard dose of “sada" than those of the control group, suggesting a probable impairment of liver function due to liver cytolysis.Mean serum AST, ALT and ALP activities were significantly lower (P<0.05) in rats that were orally administered with high dose of “magani" than that of the control group, suggesting a probable improvement in liver function (due to decrease in liver cytolysis). Mean serum TP, ALB and GLO levels were significantly higher (P<0.05) in rats that were orally administered with the various doses of“sak", “sada" and “magani" than those of the control group. This also suggests a probable improvement in the synthetic function of the liver.Thus, some dosages of “sak" and “sada could be hepatotoxic, whereas “magani" especially at the high dose administered could have pharmacologically positive effect on the liver of the rats.

Keywords: Gadagi" tea, Liver function, Oral, Rats.

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154 Haematology and Serum Biochemical Profile of Laying Chickens Reared on Deep Litter System with or without Access to Grass or Legume Pasture under Humid Tropical Climate

Authors: E. Oke, A. O. Ladokun, J. O. Daramola, O. M. Onagbesan

Abstract:

There has been a growing interest on the effects of access to pasture on poultry health status. However, there is a paucity of data on the relative benefits of grass and legume pastures. An experiment was conducted to determine the effects of rearing systems {deep litter system (DL), deep litter with access to legumes (LP) or grass (GP) pastures} haematology and serum chemistry of ISA Brown layers. The study involved the use of two hundred and forty 12 weeks old pullets. The birds were reared until 60 weeks of age. Eighty birds were assigned to each treatment; each treatment had four replicates of 20 birds each. Blood samples (2.5 ml) were collected from the wing vein of two birds per replicate and serum chemistry and haematological parameters were determined. The results showed that there were no significant differences between treatments in all the parameters considered at 18 weeks of age. At 24 weeks old, the percentage of heterophyl (HET) in DL and LP were similar but higher than that of GP. The ratio of H:L was higher (P<0.05) in DL than those of LP and GP while LP and GP were comparable. At week 38 of age, the percentage of PCV in the birds in LP and GP were similar but the birds in DL had significantly lower level than that of GP. In the early production phase, serum total protein of the birds in LP was similar to that of GP but higher (P<0.05) than that of DL. At the peak production phase (week 38), the total protein in GP and DL were similar but significantly lower than that of LP. The albumin level in LP was greater (P<0.05) than GP but similar to that of DL. In the late production phase, the total protein in LP was significantly higher than that of DL but similar to that of GP. It was concluded that rearing chickens in either grass or legume pasture did not have deleterious effects on the health of laying chickens but improved some parameters including blood protein and HET/lymphocyte.

Keywords: Rearing systems, Stylosanthes, Cynodon serum chemistry, haematology, hen.

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153 Cissampelos capensis Rhizome Extract Induces Intracellular ROS Production, Capacitation and DNA Fragmentation in Human Spermatozoa

Authors: S. Shalaweh, P. Bouic, F. Weitz, R. Henkel

Abstract:

More than 3000 plants of notable phyto-therapeutic value grow in South Africa; these include Cissampelos capensis, commonly known in Afrikaans as dawidjie or dawidjiewortel. C. capensis is the most significant and popular medicinal plant used by the Khoisan as well as other rural groups in the Western region of South Africa. Its rhizomes are traditionally used to treat male fertility problems. Yet, no studies have investigated the effects of this plant or its extracts on human spermatozoa. Therefore, this study aimed at investigating the effects of C. capensis rhizome extract (CRE) fractions on ejaculated human spermatozoa in vitro. Spermatozoa from a total of 77 semen samples were washed with human tubular fluid medium supplemented with bovine serum albumin (HTF-BSA) and incubated for 2 hours with 20 μg/ml progesterone (P4) followed by incubation with different concentrations (0, 0.05, 0.5, 5, 50, 200 μg/ml) of fractionated CRE (F1=0% MeOH, F2=30% MeOH, F3=60% MeOH and F4=100% MeOH) for 1.5 hours at 37°C. A sample without addition of CRE fractions served as control. Samples were analyzed for sperm motility, reactive oxygen species (ROS), DNA-fragmentation, acrosome reaction and capacitation. Results showed that F1 resulted in significantly higher values for ROS, capacitation and hyper-activation compared to F2, F3, and F4 with P4-stimulated samples generally having higher values. No significant effect was found for the other parameters. In conclusion, alkaloids present in F1 of CRE appear to have triggered sperm intrinsic ROS production leading to sperm capacitation and acrosome reaction induced by P4.

Keywords: Capacitation, acrosome reaction, Cissampelos capensis, DNA fragmentation, ROS.

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152 Liquid Chromatography Microfluidics for Detection and Quantification of Urine Albumin Using Linear Regression Method

Authors: Patricia B. Cruz, Catrina Jean G. Valenzuela, Analyn N. Yumang

Abstract:

Nearly a hundred per million of the Filipino population is diagnosed with Chronic Kidney Disease (CKD). The early stage of CKD has no symptoms and can only be discovered once the patient undergoes urinalysis. Over the years, different methods were discovered and used for the quantification of the urinary albumin such as the immunochemical assays where most of these methods require large machinery that has a high cost in maintenance and resources, and a dipstick test which is yet to be proven and is still debated as a reliable method in detecting early stages of microalbuminuria. This research study involves the use of the liquid chromatography concept in microfluidic instruments with biosensor as a means of separation and detection respectively, and linear regression to quantify human urinary albumin. The researchers’ main objective was to create a miniature system that quantifies and detect patients’ urinary albumin while reducing the amount of volume used per five test samples. For this study, 30 urine samples of unknown albumin concentrations were tested using VITROS Analyzer and the microfluidic system for comparison. Based on the data shared by both methods, the actual vs. predicted regression were able to create a positive linear relationship with an R2 of 0.9995 and a linear equation of y = 1.09x + 0.07, indicating that the predicted values and actual values are approximately equal. Furthermore, the microfluidic instrument uses 75% less in total volume – sample and reagents combined, compared to the VITROS Analyzer per five test samples.

Keywords: Chronic kidney disease, microfluidics, linear regression, VITROS analyzer, urinary albumin.

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151 Identification of Binding Proteins That Interact with BVDV E2 Protein in Bovine Trophoblast Cell

Authors: Yan Ren, Fei Guo, Jun Qiao, Shengwei Hu, Hui Zhang, Yuanzhi Wang, Pengyan Wang, Jinliang Sheng, Xinli Gu, Xiaojun Liu, Chuangfu Chen

Abstract:

Bovine viral diarrhea virus (BVDV) can cause lifelong persistent infection. One reason for the phenomena is attributed to BVDV infection to placenta tissue. However the mechanisms that BVDV invades into placenta tissue remain unclear. To clarify the molecular mechanisms, we investigated the possible means that BVDV entered into bovine trophoblast cells (TPC). Yeast two-hybrid system was used to identify proteins extracted from TPC, which interact with BVDV envelope glycoprotein E2. A PGbkt7-E2 yeast expression vector and TPC cDNA library were constructed. Through two rounds of screening, three positive clones were identified. Sequencing analysis indicated that all the three positive clones encoded the same protein clathrin. Physical interaction between clathrin and BVDV E2 protein was further confirmed by coimmunoprecipitation experiments. This result suggested that the clathrin might play a critical role in the process of BVDV entry into placenta tissue and might be a novel antiviral target for preventing BVDV infection.

Keywords: Bovine viral diarrhea virus, clathrin, glycoprotein E2, yeast two-hybrid system.

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150 Evaluation of the Possible Effect of Gender, Age and Duration of Diabetes on the Serum Zinc Levels of Diabetic Patients in Murzuk Area-Libya

Authors: Mukhtar H. Hassan, Muhammed A. Basher, Elhadi E. Saad, Almahdi M. Almahdi

Abstract:

The aim of this study was to demonstrate the possible effect of some variables such as age, gender, blood sugar level, and duration of diabetes on the serum level of zinc in diabetic individuals from Murzuk area. Serum zinc (Zn), Fasting blood sugar (FBS), hemoglobin HbA1c (HbA1c) were evaluated in 46 type I diabetic subjects (group 1), 48 type II diabetic subjects (group 2) and 43 healthy individuals (control) of both genders aged (30-81) years. Data showed that both diabetic groups have significantly higher (P<0.05) serum levels of Zn, FBS and HbA1c compared with controls. No significant (p>0.05) differences in serum Zn levels were observed between Males and Females. Serum Zn levels were non-significantly decreased with increasing age. In type II diabetic subjects, serum Zn levels were non-significantly decreased with increasing duration of disease whereas those in type I were non-significantly increased.

Keywords: Blood sugar, diabetes, HbA1c, zinc.

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149 Gamma Glutamyl Transferase and Lactate Dehydrogenase as Biochemical Markers of Severity of Preeclampsia

Authors: S. M. Munde, N. R. Hazari, A. P. Thorat, S. B. Gaikwad, V. S. Hatolkar

Abstract:

This study was conducted to examine the possible role of serum Gamma-glutamyltransferase (GGT) and Lactate dehydrogenase (LDH) in the prediction of severity of preeclampsia. The study group comprised of 40 preeclamptic cases (22 with mild and 18 with severe) and 40 healthy normotensive pregnant controls. Serum samples of all the cases were assayed for GGT and LDH. Demographic, hemodynamic and laboratory data as well as serum GGT and LDH levels were compared among the three groups.

The results indicated that severe preeclamptic cases had significantly increased levels of serum GGT and LDH. The symptoms in severe preeclamptic women were significantly increased in patients with GGT > 70 IU/L and LDH >800 IU/L. Elevated levels of serum GGT and LDH can be used as biochemical markers which reflects the severity of preeclampsia and useful for the management of preeclampsia to decrease maternal and fetal morbidity and mortality.

Keywords: Severe Preeclampsia, GGT, LDH.

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148 In vitro Effects of Berberine on the Vitality and Oxidative Profile of Bovine Spermatozoa

Authors: Eva Tvrdá, Hana Greifová, Peter Ivanič, Norbert Lukáč

Abstract:

The aim of this study was to evaluate the dose- and time-dependent in vitro effects of berberine (BER), a natural alkaloid with numerous biological properties on bovine spermatozoa during three time periods (0 h, 2 h, 24 h). Bovine semen samples were diluted and cultivated in physiological saline solution containing 0.5% DMSO together with 200, 100, 50, 10, 5, and 1 μmol/L BER. Spermatozoa motility was assessed using the computer assisted semen analyzer. The viability of spermatozoa was assessed by the metabolic (MTT) assay, production of superoxide radicals was quantified using the nitroblue tetrazolium (NBT) test, and chemiluminescence was used to evaluate the generation of reactive oxygen species (ROS). Cell lysates were prepared and the extent of lipid peroxidation (LPO) was evaluated using the TBARS assay. The results of the movement activity showed a significant increase in the motility during long term cultivation in case of concentrations ranging between 1 and 10 μmol/L BER (P < 0.01; P < 0.001; 24 h). At the same time, supplementation of 1, 5 and 10 μmol/L BER led to a significant preservation of the cell viability (P < 0.001; 24 h). BER addition at a range of 1-50 μmol/L also provided a significantly higher protection against superoxide (P < 0.05) and ROS (P < 0.001; P < 0.01) overgeneration as well as LPO (P < 0.01; P<0.05) after a 24 h cultivation. We may suggest that supplementation of BER to bovine spermatozoa, particularly at concentrations ranging between 1 and 50 μmol/L, may offer protection to the motility, viability and oxidative status of the spermatozoa, particularly notable at 24 h.

Keywords: Berberine, bulls, motility, oxidative profile, spermatozoa, viability.

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147 Serum Nitric Oxide and Sialic Acid: Possible Biochemical Markers for Progression of Diabetic Nephropathy

Authors: Syed M. Shahid, Rozeena Shaikh, Syeda N. Nawab, Shah A. Qader, Abid Azhar, Tabassum Mahboob

Abstract:

This study was designed to investigate the role of serum nitric oxide and sialic acid in the development of diabetic nephropathy as disease marker. Total 210 diabetic patients (age and sex matched) were selected followed by informed consent and divided into four groups (70 each) as I: control; II: diabetic; III: diabetic hypertensive; IV: diabetic nephropathy. The blood samples of all subjects were collected and analyzed for serum nitric oxide, sialic acid, fasting blood glucose, serum urea, creatinine, HbA1c and GFR. The BMI, systolic and diastolic blood pressures, blood glucose, HbA1c and serum sialic acid levels were high (p<0.01) in group II as compared to control subjects. The higher levels (p<0.01) of BMI, systolic and diastolic blood pressures, blood glucose, HbA1c, serum urea, creatinine and sialic acid were observed in group III and IV as compared to controls. Significantly low levels of GFR and serum nitric oxide (p<0.01) were observed in group III and IV as compared to controls. Results indicated that serum nitric oxide and sialic acid are the major biochemical indicators for micro and macrovascular complications of diabetes such as hypertension and nephropathy. These should be taken into account during screening procedures regarding identifications of the diabetic patients to get them rid of progressive renal impairment to ESRD.

Keywords: Diabetic nephropathy, hypertension, nitric oxide, sialic acid.

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146 Histochemistry of Intestinal Enzymes of Juvenile Dourado Salminus brasiliensis Fed Bovine Colostrum

Authors: Debora B. Moretti, Wiolene M. Nordi, Thaline Maira P. Cruz, José Eurico P. Cyrino, Raul Machado-Neto

Abstract:

Enzyme activity was evaluated in the intestine of juvenile dourado (Salminus brasiliensis) fed with diets containing 0, 10 or 20% of lyophilized bovine colostrum (LBC) inclusion for either 30 or 60 days. The intestinal enzymes acid and alkaline phosphatase (ACP and ALP, respectively), non-specific esterase (NSE), lipase (LIP), dipeptidyl aminopeptidase IV (DAP IV) and leucine aminopeptidase (LAP) were studied using histochemistry in four intestinal segments (S1, S2, S3 and posterior intestine). Weak proteolitic activity was observed in all intestinal segments for DAP IV and LAP. The activity of NSE and LIP was also weak in all intestines, except for the moderate activity of NSE in the S2 of 20% LBC group after 30 days and in the S1 of 0% LBC group after 60 days. The ACP was detected only in the S2 and S3 of the 10% LBC group after 30 days. Moderate and strong staining was observed in the first three intestinal segments for ALP and weak activity in the posterior intestine. The activity of DAP IV, LAP and ALP were also present in the cytoplasm of the enterocytes. In the present results, bovine colostrum feeding did not cause alterations in activity of intestinal enzymes.

Keywords: Carnivorous fish, enterocyte, intestinal epithelium, teleost.

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145 Quantitative Analysis of Carcinoembryonic Antigen (CEA) Using Micromechanical Piezoresistive Cantilever

Authors: Meisam Omidi, M. Mirijalili, Mohammadmehdi Choolaei, Z. Sharifi, F. Haghiralsadat, F. Yazdian

Abstract:

In this work, we have used arrays of micromechanical piezoresistive cantilever with different geometries to detect carcinoembryonic antigen (CEA), which is known as an important biomarker associated with various cancers such as colorectal, lung, breast, pancreatic, and bladder cancer. The sensing principle is based on the surface stress changes induced by antigen–antibody interaction on the microcantilevers surfaces. Different concentrations of CEA in a human serum albumin (HSA) solution were detected as a function of deflection of the beams. According to the experiments, it was revealed that microcantilevers have surface stress sensitivities in the order of 8 (mJ/m). This matter allows them to detect CEA concentrations as low as 3 ng/mL or 18 pM. This indicates the fact that the self-sensing microcantilevers approach is beneficial for pathological tests.

Keywords: Micromechanical biosensors, Carcinoembryonic antigen (CEA), surface stress.

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144 Contrast-Enhanced Magnetic Resonance Angiography in Rats with Gadobenate Dimeglumine at 3T

Authors: Jo-Chi Jao, Yen-Ku Chen, Twei-Shiun Jaw, Po-Chou Chen

Abstract:

This study aimed to investigate the magnetic resonance (MR) signal enhancement ratio (ER) of contrast-enhanced MR angiography (CE-MRA) in normal rats with gadobenate dimeglumine (Gd-BOPTA) using a clinical 3T scanner and an extremity coil. The relaxivities of Gd-BOPTA with saline only and with 4.5% human serum albumin (HSA) were also measured. Compared with Gadolinium diethylenetriaminepentaacetic acid (Gd-DTPA), Gd-BOPTA had higher relaxivities. The maximum ER of aorta (ERa), kidney, liver and muscle with Gd-BOPTA were higher than those with Gd-DTPA. The maximum ERa appeared at 1.2 min and decayed to half at 10 min after Gd-BOPTA injection. This information is helpful for the design of CE-MRA study of rats.

Keywords: Contrast-Enhanced Magnetic Resonance Angiography, Gd-BOPTA, Gd-DTPA, Rat.

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143 Effect of Cold Plasma-Surface Modification on Surface Wettability and Initial Cell Attachment

Authors: Masao Yoshinari, Jianhua Wei, Kenichi Matsuzaka, Takashi Inoue

Abstract:

A thin coating of hexamethyldisiloxane and subsequent O2-plasma treatment was performed on mirror-polished titanium in order to regulate the wide range of wettability including 106 and almost 0 degrees of contact angles. The adsorption behavior of fibronectin and albumin in both individual and competitive mode, and initial attachment of fibroblasts and osteoblasts were investigated. Individually, fibronectin adsorption showed a biphasic inclination, whereas albumin showed greater adsorption to hydrophobic surfaces. In competitive mode, in solution containing both fibronectin and albumin, fibronectin showed greater adsorption on hydrophilic surfaces, whereas Alb predominantly adsorbed on hydrophobic surfaces. Initial attachment of both cells increased with increase in surface wettability, in particular, on super-hydrophilic surface, which correlated well with fibronectin adsorption in competitive mode. These results suggest that a cold plasma-surface modification enabled to regulate the surface wettability, and fibronectin adsorption may be responsible for increasing cell adhesion on hydrophilic surfaces in a body fluid

Keywords: cold plasma-surface modification, wettability, protein adsorption, initial cell attachment.

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142 Biosensor Measurement of Urea Coonncentration in Human Blood Serum

Authors: O. L. Kukla, S. V. Marchenko, O. A. Zinchenko, O. S. Pavluchenko, O. M. KKuukla, S. V. Dzyadevych, O. P. Soldatkin

Abstract:

An application of the highly biosensor based on pH-sensitive field immobilized urease for urea analysis was demo The main analytical characteristics of the bios determined; the conditions of urea measureme blood were optimized. A conceptual possibility biosensor for detection of urea concentratio patients suffering from renal insufficiency was sensitive and selective effect transistor and monstrated in this work. iosensor developed were ment in real samples of ility of application of the tion in blood serum of as shown.

Keywords: Biosensor, blood serum, pH transistor, urea, urease, field-effect

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141 Protective Effect of L-Carnitine against Gentamicin-Induced Nephrotoxicity in Rats

Authors: Mohamed F. Ahmed, Mabruka S. Elashheb, Fatma M. Ben Rabha

Abstract:

This study aimed to determine the possible protective effects of L‐carnitine against gentamicin‐induced nephrotoxicity. Forty male albino rats were divided into 4 groups (10 rats each); Group 1: normal control, group 2: induced nephrotoxicity (gentamicin 50 mg/kg/day S.C; 8 days), group 3: treated with L‐ carnitine (40 mg/kg/d SC for 12 days) and group 4: treated with L‐ carnitine 4 days before and for 8 days in concomitant with gentamicin. Gentamicin‐induced nephrotoxicity (group 2): caused significant increase in serum urea, creatinine, urinary N‐acetyl‐B‐D‐ glucosaminidase (NAG), gamma glutamyl transpeptidase (GGT), urinary total protein and kidney tissue malondialdehyde (MDA) with significant decrease in serum superoxide dismutase (SOD), serum catalase and creatinine clearance and marked tubular necrosis in the proximal convoluted tubules with interruption in the basement membrane around the necrotic tubule compared to the normal control group. L‐carnitine 4 days before and for 8 days in concomitant with gentamicin (group 4) offered marked decrease in serum urea, serum creatinine, urinary NAG, urinary GGT, urinary proteins and kidney tissue MDA, with marked increase in serum SOD, serum catalase and creatinine clearance with marked improvement in the tubular damage compared to gentamicin‐induced nephrotoxicity group. L‐carnitine administered for 12 days produced no change in the parameters mentioned above as compared to the normal control group. In conclusion: L‐carnitine could reduce most of the biochemical parameters and also improve the histopathological features of kidney asscociated with gentamicin induced‐nephrotoxicity. 

Keywords: Gentamicin, kidney disease, L‐carnitine, nephrotoxicity.

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140 Molecular Epidemiology and Genotyping of Bovine Viral Diarrhea Virus in Xinjiang Uygur Autonomous Region of China

Authors: Yan Ren, Jun Qiao, Xianxia Liu, Pengyan Wang, Qiang Fu, Huijun Shi, Fei Guo, Yuanzhi Wang, Hui Zhang, Jinliang Sheng, Xinli Gu, Xiao-Jun Liu, Chuangfu Chen

Abstract:

As part of national epidemiological survey on bovine viral diarrhea virus (BVDV), a total of 274 dejecta samples were collected from 14 cattle farms in 8 areas of Xinjiang Uygur Autonomous Region in northwestern China. Total RNA was extracted from each sample, and 5--untranslated region (UTR) of BVDV genome was amplified by using two-step reverse transcriptase-polymerase chain reaction (RT-PCR). The PCR products were subsequently sequenced to study the genetic variations of BVDV in these areas. Among the 274 samples, 33 samples were found virus-positive. According to sequence analysis of the PCR products, the 33 samples could be arranged into 16 groups. All the sequences, however, were highly conserved with BVDV Osloss strains. The virus possessed theses sequences belonged to BVDV-1b subtype by phylogenetic analysis. Based on these data, we established a typing tree for BVDV in these areas. Our results suggested that BVDV-1b was a predominant subgenotype in northwestern China and no correlation between the genetic and geographical distances could be observed above the farm level.

Keywords: bovine viral diarrhea virus, molecular epidemiology, phylogenetic analysis.

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139 The Alterations of Some Pancreas Gland Hormones after an Aerobic Strenuous Exercise in Male Students

Authors: M. Javad Pourvaghar, A. Reza Shahsavar

Abstract:

The alterations in pancreas gland secretion hormones following an aerobic and exhausting exercise was the purpose of this study. Sixteen healthy men participated in the study. The blood samples of these participants were taken in four stages under fasting condition. The first sample was taken before Bruce exhausting and aerobic test, the second sample was taken after Bruce exercise and the third and forth stages samples were taken 24 and 48 hours after the exercises respectively. The final results indicated that a strenuous aerobic exercise can have a significant effect on glucagon and insulin concentration of blood serum. The increase in blood serum insulin was higher after 24 and 48 hours. It seems that an intensive exercise has little effect on changes in glucagon concentration of blood serum. Also, disorder in secretion in glucagon and insulin concentration of serum disturbs athletes- exercise.

Keywords: Intensive Exercise, Bruce Protocol, Glucagon, Insulin

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