Search results for: serum xanthine oxidase
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 1089

Search results for: serum xanthine oxidase

519 Anti-Apoptotic Effect of Pueraria tuberosa in Rats with Streptozotocin Induced Diabetic Nephropathy

Authors: Rashmi Shukla, Yamini Bhusan Tripathi

Abstract:

Diabetic nephropathy (DN) is characterized as diabetic kidney disease which involves many pathways e.g. hyperactivated protein kinase c (PKC), polyol pathway, excess production of advanced glycation end product (AGEs) & free radical accumulation etc. All of them results to hypoxia followed by apoptosis of podocytes, glomerulosclerosis, extracellular matrix (ECM) accumulation and fibrosis resulting to irreversible changes in kidney. This is continuously rising worldwide and there are not enough specific drugs, to retard its progress. Due to increasing side effects of allopathic drugs, interest in herbal remedies is growing. Earlier, we have reported that PTY-2 (a phytomedicine, derived from Pueraria tuberosa Linn.) inhibits the accumulation of extracellular matrix (ECM) through activation of MMP-9. Present study exhibited the therapeutic potential of Pueraria tuberosa in the prevention of podocytes apoptosis and modulation of nephrin expression in streptozotocin (STZ) induced DN rats. DN rats were produced by maintaining persistent hyperglycemia for 8 weeks by intra-peritoneal injection of 55 mg/kg streptozotocin (STZ). These rats were randomly divided in 2 groups, i.e. DN control, and DN+ water extract of Pueraria tuberosa (PTW). One group of age-matched normal rats served as non-diabetic control (group-1), The STZ induced DN rats (group-2) and DN+PTW treated rats (group-3). The PTW was orally administered (0.3g/kg) daily to group-2 rats and drug vector (1 ml of 10% tween 20) in control rats. The treatments were continued for 20 days and blood and urine samples were collected. Rats were then sacrificed to investigate the expression Bcl2, Bax and nephroprotective protein i.e. nephrin in kidney glomerulus. The effect of PTW was evaluated, we have found that the PTW significantly(p < .001) reversed the raised serum urea, serum creatinine, urine protein and improved the creatinine clearance in STZ induce diabetic nephropathy in rats and also significantly(p < .001) prevented the rise in urine albumin excretion. The Western blot analysis of kidney tissue homogenate showed increased expression of Bcl2 in PTW treated rats. The RT-PCR showed the increased expression and accumulation of nephrin mRNA. The confocal photomicrographs also supported the reduction of Bax and a simultaneous increase in Bcl2 and nephrin in glomerular podocytes. Hence, our finding suggests that the nephroprotective role of PTW is mediated via restoration of nephrin thus prevents the podocytes apoptosis and ameliorates diabetic nephropathy. The clinical trial of PTW would prove to be a potential food supplement/ drug of alternative medicine for patients with diabetic nephropathy in early stage.

Keywords: Pueraria tuberosa, diabetic nephropathy, anti-apoptosis, nephrin

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518 Aloe vera Prevents Injuries Induced by Whole Body X-ray Irradiation in Rodents

Authors: Shashi Bala, Neha A. Chugh, Subhash C. Bansal, Mohal L. Garg, Ashwani Koul

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Purpose: The present study was designed to evaluate the radioprotective efficacy of Aloe vera from whole body X-ray exposure in rodents. Materials and Methods: For this purpose, after on week’s acclimatization, male balb/c mice procured from Central Animal House, Panjab University, Chandigarh (India), were divided into four groups: Group I mice served as control. Group II mice were orally administrated Aloe vera pulp extract (50 mg/ kg body weight) on alternate days for 30 days. Group III mice were subjected to whole body X-ray irradiation to cumulative dose of 2Gy (0.258Gy twice a day for four days in the last week). Group IV animals were pretreated with Aloe vera pulp extract on alternate days as in Group II and in the last week of the study, they were exposed to X-ray as in Group III. Results: Spleen of X-ray irradiated mice showed histopathological alterations accompanied with enhanced activity of lactate dehydrogenase (LDH) in serum. Elevated levels of reactive oxygen species (ROS), lipid peroxidation (LPO), enhanced activities in Glutathione based enzymes such as Glutathione peroxidase (GSH-Px), Glutathione reductase (GR), Catalase (CAT), Superoxide dismutase (SOD) associated with depletion in reduced Glutathione (GSH) concentration were observed after X-ray exposure in blood plasma and spleen.. Pro-inflammatory cytokines like tumor necrosis factors (TNF-α) and Inteleukin-6 (IL-6) levels were also found to be enhanced in serum of irradiated mice. Irradiation-induced significant elevation in Total leucocyte counts (TLC), neutrophil counts and decline in platelet counts, associated with unaltered levels of red blood cell counts (RBC’s) and haemoglobin (Hb) in various treatment groups. Clastogenic damage and apoptosis was also found to be increase in splenic tissue of X-ray exposed mice as assessed by micronucleus and TUNEL assay. However, X-ray irradiated animals administered with Aloe vera revealed significant improvement in levels of ROS/ LPO, LDH activity, and antioxidant mechanism. Aloe vera pretreated animals exhibited less severe damage, and early recovery in micronucleated cells, hematological parameters, apoptotic cells and inflammatory markers as compared to X-ray exposed mice. Conclusion: These results indicate that the radioprotective potential of Aloe vera against X-ray induced damage. This may be due to its free radical scavenging, antioxidant, anti-apoptotic and anti-inflammatory properties.

Keywords: aloe vera, antioxidant defense system, lactate dehydrogenase (LDH), micronucleus assay, x-ray

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517 Electrophoretic Deposition of Ultrasonically Synthesized Nanostructured Conducting Poly(o-phenylenediamine)-Co-Poly(1-naphthylamine) Film for Detection of Glucose

Authors: Vaibhav Budhiraja, Chandra Mouli Pandey

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The ultrasonic synthesis of nanostructured conducting copolymer is an effective technique to synthesize polymer with desired chemical properties. This tailored nanostructure, shows tremendous improvement in sensitivity and stability to detect a variety of analytes. The present work reports ultrasonically synthesized nanostructured conducting poly(o-phenylenediamine)-co-poly(1-naphthylamine) (POPD-co-PNA). The synthesized material has been characterized using Fourier transform infrared spectroscopy (FTIR), ultraviolet-visible spectroscopy, transmission electron microscopy, X-ray diffraction and cyclic voltammetry. FTIR spectroscopy confirmed random copolymerization, while UV-visible studies reveal the variation in polaronic states upon copolymerization. High crystallinity was achieved via ultrasonic synthesis which was confirmed by X-ray diffraction, and the controlled morphology of the nanostructures was confirmed by transmission electron microscopy analysis. Cyclic voltammetry shows that POPD-co-PNA has rather high electrochemical activity. This behavior was explained on the basis of variable orientations adopted by the conducting polymer chains. The synthesized material was electrophoretically deposited at onto indium tin oxide coated glass substrate which is used as cathode and parallel platinum plate as the counter electrode. The fabricated bioelectrode was further used for detection of glucose by crosslinking of glucose oxidase in the PODP-co-PNA film. The bioelectrode shows a surface-controlled electrode reaction with the electron transfer coefficient (α) of 0.72, charge transfer rate constant (ks) of 21.77 s⁻¹ and diffusion coefficient 7.354 × 10⁻¹⁵ cm²s⁻¹.

Keywords: conducting, electrophoretic, glucose, poly (o-phenylenediamine), poly (1-naphthylamine), ultrasonic

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516 Localized Detection of ᴅ-Serine by Using an Enzymatic Amperometric Biosensor and Scanning Electrochemical Microscopy

Authors: David Polcari, Samuel C. Perry, Loredano Pollegioni, Matthias Geissler, Janine Mauzeroll

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ᴅ-serine acts as an endogenous co-agonist for N-methyl-ᴅ-aspartate receptors in neuronal synapses. This makes it a key component in the development and function of a healthy brain, especially given its role in several neurodegenerative diseases such as Alzheimer’s disease and dementia. Despite such clear research motivations, the primary site and mechanism of ᴅ-serine release is still currently unclear. For this reason, we are developing a biosensor for the detection of ᴅ-serine utilizing a microelectrode in combination with a ᴅ-amino acid oxidase enzyme, which produces stoichiometric quantities of hydrogen peroxide in response to ᴅ-serine. For the fabrication of a biosensor with good selectivity, we use a permselective poly(meta-phenylenediamine) film to ensure only the target molecule is reacted, according to the size exclusion principle. In this work, we investigated the effect of the electrodeposition conditions used on the biosensor’s response time and selectivity. Careful optimization of the fabrication process allowed for enhanced biosensor response time. This allowed for the real time sensing of ᴅ-serine in a bulk solution, and also provided in means to map the efflux of ᴅ-serine in real time. This was done using scanning electrochemical microscopy (SECM) with the optimized biosensor to measure localized release of ᴅ-serine from an agar filled glass capillary sealed in an epoxy puck, which acted as a model system. The SECM area scan simultaneously provided information regarding the rate of ᴅ-serine flux from the model substrate, as well as the size of the substrate itself. This SECM methodology, which provides high spatial and temporal resolution, could be useful to investigate the primary site and mechanism of ᴅ-serine release in other biological samples.

Keywords: ᴅ-serine, enzymatic biosensor, microelectrode, scanning electrochemical microscopy

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515 Isolation and Characterization of an Ethanol Resistant Bacterium from Sap of Saccharum officinarum for Efficient Fermentation

Authors: Rukshika S Hewawasam, Sisira K. Weliwegamage, Sanath Rajapakse, Subramanium Sotheeswaran

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Bio fuel is one of the emerging industries around the world due to arise of crisis in petroleum fuel. Fermentation is a cost effective and eco-friendly process in production of bio-fuel. So inventions in microbes, substrates, technologies in fermentation cause new modifications in fermentation. One major problem in microbial ethanol fermentation is the low resistance of conventional microorganisms to the high ethanol concentrations, which ultimately lead to decrease in the efficiency of the process. In the present investigation, an ethanol resistant bacterium was isolated from sap of Saccharum officinarum (sugar cane). The optimal cultural conditions such as pH, temperature, incubation period, and microbiological characteristics, morphological characteristics, biochemical characteristics, ethanol tolerance, sugar tolerance, growth curve assay were investigated. Isolated microorganism was tolerated to 18% (V/V) of ethanol concentration in the medium and 40% (V/V) glucose concentration in the medium. Biochemical characteristics have revealed as Gram negative, non-motile, negative for Indole test ,Methyl Red test, Voges- Proskauer`s test, Citrate Utilization test, and Urease test. Positive results for Oxidase test was shown by isolated bacterium. Sucrose, Glucose, Fructose, Maltose, Dextrose, Arabinose, Raffinose, Lactose, and Sachcharose can be utilized by this particular bacterium. It is a significant feature in effective fermentation. The fermentation process was carried out in glucose medium under optimum conditions; pH 4, temperature 30˚C, and incubated for 72 hours. Maximum ethanol production was recorded as 12.0±0.6% (V/V). Methanol was not detected in the final product of the fermentation process. This bacterium is especially useful in bio-fuel production due to high ethanol tolerance of this microorganism; it can be used to enhance the fermentation process over conventional microorganisms. Investigations are currently conducted on establishing the identity of the bacterium

Keywords: bacterium, bio-fuel, ethanol tolerance, fermentation

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514 Helicobacter Pylori Detection by Invasive and Noninvasive Diagnostic Tests from Dyspepsia Patients

Authors: Muhammad Suhail Ibrahim, Ahmad Mujtaba

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Background: The accuracy of the most frequently used tests for diagnosing Helicobacter pylori is always under consideration in clinical settings. A reliable diagnosis is crucial to confirm the success of therapy. Objective: The aim of this research was to study the isolation frequency of H. pylori from patients compatible with gastritis or gastric ulcer and to compare some feasible non-invasive and invasive methods for the diagnosis of infection. Materials and Methods: Ninety-six gastric biopsy and blood samples were obtained with various gastroduodenal symptoms after obtaining informed consent. The biopsies were analyzed and compared using the culture, microscopic examination, histopathology, Rapid urease RUT), serology, biochemical, antibiotic susceptibility test and molecular method. Results: A number of 40 (41.67%) were considered H. pylori positive in both histopathology and RUT. On the other hand, 46 patients were positive against anti IgA and IgG by ELISA. Eighteen biopsies were positive according to the culture test. This was further confirmed by endoscopic examination, urease, catalase and oxidase tests. A high percentage of resistance to polymyxin B, amoxicillin, and kanamycin was observed (100, 88.89, and 77.78%, respectively). A gene (Cag A) was also detected by using molecular technique which appeared positive in 16 patients. The sensitivity/specificity (%) of diagnostic method was 95/77 for histology, 100/83.5 for rapid urease, 85.7/90 for gram staining, 100/66.6 for IgG serology, 100/79.5 for IgA serology, 100/75.0 for PCR, 100/79.04 for combination of RUT and IgG serology and 100/92.4 for combination of RUT, gram staining and IgG serology. Conclusion: In view of the result obtained, PCR appeared to be the most reliable test. However, higher sensitivity and specificity were also recorded for other tests. So, for more accurate results, it is advisable not to rely solely on a single method for detection.

Keywords: helicobacter pylori, isolation, detection, culture, urease, polymerase chain reaction, antibiotic susceptibility test, dyspeptic patients

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513 Enzyme Producing Psyhrophilic Pseudomonas app. Isolated from Poultry Meats

Authors: Ali Aydin, Mert Sudagidan, Aysen Coban, Alparslan Kadir Devrim

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Pseudomonas spp. (specifically, P. fluorescens and P. fragi) are considered the principal spoilage microorganisms of refrigerated poultry meats. The higher the level psychrophilic spoilage Pseudomonas spp. on carcasses at the end of processing lead to decrease the shelf life of the refrigerated product. The aim of the study was the identification of psychrophilic Pseudomonas spp. having proteolytic and lipolytic activities from poultry meats by 16S rRNA and rpoB gene sequencing, investigation of protease and lipase related genes and determination of proteolytic activity of Pseudomonas spp. In the of isolation procedure, collected chicken meat samples from local markets and slaughterhouses were homogenized and the lysates were incubated on Standard method agar and Skim Milk agar for selection of proteolytic bacteria and tributyrin agar for selection of lipolytic bacteria at +4 °C for 7 days. After detection of proteolytic and lipolytic colonies, the isolates were firstly analyzed by biochemical tests such as Gram staining, catalase and oxidase tests. DNA gene sequencing analysis and comparison with GenBank revealed that 126 strong enzyme Pseudomonas spp. were identified as predominantly P. fluorescens (n=55), P. fragi (n=42), Pseudomonas spp. (n=24), P. cedrina (n=2), P. poae (n=1), P. koreensis (n=1), and P. gessardi (n=1). Additionally, protease related aprX gene was screened in the strains and it was detected in 69/126 strains, whereas, lipase related lipA gene was found in 9 Pseudomonas strains. Protease activity was determined using commercially available protease assay kit and 5 strains showed high protease activity. The results showed that psychrophilic Pseudomonas strains were present in chicken meat samples and they can produce important levels of proteases and lipases for food spoilage to decrease food quality and safety.

Keywords: Pseudomonas, chicken meat, protease, lipase

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512 Comparison of Two Methods of Cryopreservation of Testicular Tissue from Prepubertal Lambs

Authors: Rensson Homero Celiz Ygnacio, Marco Aurélio Schiavo Novaes, Lucy Vanessa Sulca Ñaupas, Ana Paula Ribeiro Rodrigues

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The cryopreservation of testicular tissue emerges as an alternative for the preservation of the reproductive potential of individuals who still cannot produce sperm; however, they will undergo treatments that may affect their fertility (e.g., chemotherapy). Therefore, the present work aims to compare two cryopreservation methods (slow freezing and vitrification) in testicular tissue of prepubertal lambs. For that, to obtain the testicular tissue, the animals were castrated and the testicles were collected immediately in a physiological solution supplemented with antibiotics. In the laboratory, the testis was split into small pieces. The total size of the testicular fragments was 3×3x1 mm³ and was placed in a dish contained in Minimum Essential Medium (MEM-HEPES). The fragments were distributed randomly into non-cryopreserved (fresh control), slow freezing (SF), and vitrified. To SF procedures, two fragments from a given male were then placed in a 2,0 mL cryogenic vial containing 1,0 mL MEM-HEPES supplemented with 20% fetal bovine serum (FBS) and 20% dimethylsulfoxide (DMSO). Tubes were placed into a Mr. Frosty™ Freezing container with isopropyl alcohol and transferred to a -80°C freezer for overnight storage. On the next day, each tube was plunged into liquid nitrogen (NL). For vitrification, the ovarian tissue cryosystem (OTC) device was used. Testicular fragments were placed in the OTC device and exposed to the first vitrification solution composed of MEM-HEPES supplemented with 10 mg/mL Bovine Serum Albumin (BSA), 0.25 M sucrose, 10% Ethylene glycol (EG), 10% DMSO and 150 μM alpha-lipoic acid for four min. The VS1 was discarded and then the fragments were submerged into a second vitrification solution (VS2) containing the same composition of VS1 but 20% EG and 20% DMSO. VS2 was then discarded and each OTC device containing up to four testicular fragments was closed and immersed in NL. After the storage period, the fragments were removed from the NL, kept at room temperature for one min and then immersed at 37 °C in a water bath for 30 s. Samples were warmed by sequentially immersing in solutions of MEM-HEPES supplemented with 3 mg/mL BSA and decreasing concentrations of sucrose. Hematoxylin-eosin staining to analyze the tissue architecture was used. The score scale used was from 0 to 3, classified with a score 0 representing normal morphologically, and 3 were considered a lot of alteration. The histomorphological evaluation of the testicular tissue shows that when evaluating the nuclear alteration (distinction of nucleoli and condensation of nuclei), there are no differences when using slow freezing with respect to the control. However, vitrification presents greater damage (p <0.05). On the other hand, when evaluating the epithelial alteration, we observed that the freezing showed scores statistically equal to the control in variables such as retraction of the basement membrane, formation of gaps and organization of the peritubular cells. The results of the study demonstrated that cryopreservation using the slow freezing method is an excellent tool for the preservation of pubertal testicular tissue.

Keywords: cryopreservation, slow freezing, vitrification, testicular tissue, lambs

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511 Ultrasensitive Hepatitis B Virus Detection in Blood Using Nano-Porous Silicon Oxide: Towards POC Diagnostics

Authors: N. Das, N. Samanta, L. Pandey, C. Roy Chaudhuri

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Early diagnosis of infection like Hep-B virus in blood is important for low cost medical treatment. For this purpose, it is desirable to develop a point of care device which should be able to detect trace quantities of the target molecule in blood. In this paper, we report a nanoporous silicon oxide sensor which is capable of detecting down to 1fM concentration of Hep-B surface antigen in blood without the requirement of any centrifuge or pre-concentration. This has been made possible by the presence of resonant peak in the sensitivity characteristics. This peak is observed to be dependent only on the concentration of the specific antigen and not on the interfering species in blood serum. The occurrence of opposite impedance change within the pores and at the bottom of the pore is responsible for this effect. An electronic interface has also been designed to provide a display of the virus concentration.

Keywords: impedance spectroscopy, ultrasensitive detection in blood, peak frequency, electronic interface

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510 Nano-Immunoassay for Diagnosis of Active Schistosomal Infection

Authors: Manal M. Kame, Hanan G. El-Baz, Zeinab A.Demerdash, Engy M. Abd El-Moneem, Mohamed A. Hendawy, Ibrahim R. Bayoumi

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There is a constant need to improve the performance of current diagnostic assays of schistosomiasis as well as develop innovative testing strategies to meet new testing challenges. This study aims at increasing the diagnostic efficiency of monoclonal antibody (MAb)-based antigen detection assays through gold nanoparticles conjugated with specific anti-Schistosoma mansoni monoclonal antibodies. In this study, several hybidoma cell lines secreting MAbs against adult worm tegumental Schistosoma antigen (AWTA) were produced at Immunology Department of Theodor Bilharz Research Institute and preserved in liquid nitrogen. One MAb (6D/6F) was chosen for this study due to its high reactivity to schistosome antigens with highest optical density (OD) values. Gold nanoparticles (AuNPs) were functionalized and conjugated with MAb (6D/6F). The study was conducted on serum samples of 116 subjects: 71 patients with S. mansoni eggs in their stool samples group (gp 1), 25 with other parasites (gp2) and 20 negative healthy controls (gp3). Patients in gp1 were further subdivided according to egg count in their stool samples into Light infection {≤ 50 egg per gram(epg) (n= 17)}, moderate {51-100 epg (n= 33)} and severe infection {>100 epg(n= 21)}. Sandwich ELISA was performed using (AuNPs -MAb) for detection of circulating schistosomal antigen (CSA) levels in serum samples of all groups and the results were compared with that after using MAb/ sandwich ELISA system. Results Gold- MAb/ ELISA system reached a lower detection limit of 10 ng/ml compared to 85 ng/ml on using MAb/ ELISA and the optimal concentrations of AuNPs -MAb were found to be 12 folds less than that of MAb/ ELISA system for detection of CSA. The sensitivity and specificity of sandwich ELISA for detection of CSA levels using AuNPs -MAb were 100% & 97.8 % respectively compared to 87.3% &93.38% respectively on using MAb/ ELISA system. It was found that CSA was detected in 9 out of 71 S.mansoni infected patients on using AuNPs - MAb/ ELISA system and was not detected by MAb/ ELISA system. All those patients (9) was found to have an egg count below 50 epg feces (patients with light infections). ROC curve analyses revealed that sandwich ELISA using gold-MAb was an excellent diagnostic investigator that could differentiate Schistosoma patients from healthy controls, on the other hand it revealed that sandwich ELISA using MAb was not accurate enough as it could not recognize nine out of 71 patients with light infections. Conclusion Our data demonstrated that: Loading gold nanoparticles with MAb (6D/6F) increases the sensitivity and specificity of sandwich ELISA for detection of CSA, thus active (early) and light infections could be easily detected. Moreover this binding will decrease the amount of MAb consumed in the assay and lower the coast. The significant positive correlation that was detected between ova count (intensity of infection) and OD reading in sandwich ELISA using gold- MAb enables its use to detect the severity of infections and follow up patients after treatment for monitoring of cure.

Keywords: Schistosomiasis, nanoparticles, gold, monoclonal antibodies, ELISA

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509 Acute Hepatotoxicity of Nano and Micro-Sized Iron Particles in Adult Albino Rats

Authors: Ghada Hasabo, Mahmoud Saber Elbasiouny, Mervat Abdelsalam, Sherin Ghaleb, Niveen Eldessouky

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In the near future, nanotechnology is envisaged for large scale use. Hence health and safety issues of nanoparticles should be promptly addressed. In the present study the acute hepatoxicity assessment due to high single oral dose of nano iron and micro iron particles were studied. The normal daily activities, biochemical alterations, blood coagulation, histopathological changes in Wister rats were the aspect of the toxicological assessment.This work found that significant alterations in biochemical enzymes (serum iron level, liver enzymes, albumin, and bilirubin levels), blood coagulation (PT, PC, INR), and histopathological changes occurred more prominently in the nano iron particle treated group.

Keywords: nanobiotechnology, nanosystems, nanomaterials, nanotechnology

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508 Applicability of Polyisobutylene-Based Polyurethane Structures in Biomedical Disciplines: Some Calcification and Protein Adsorption Studies

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

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

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

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507 Biocompatibility and Sensing Ability of Highly Luminescent Synthesized Core-Shell Quantum Dots

Authors: Mohan Singh Mehata, R. K. Ratnesh

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CdSe, CdSe/ZnS, and CdSe/CdS core-shell quantum dots (QDs) of 3-4 nm were developed by using chemical route and following successive ion layer adsorption and reaction (SILAR) methods. The prepared QDs have been examined by using X-ray diffraction, high-resolution electron microscopy and optical spectroscopy. The photoluminescence (PL) quantum yield (QY) of core-shell QDs increases with respect to the core, indicating that the radiative rate increases by the formation of shell around core, as evident by the measurement of PL lifetime. Further, the PL of bovine serum albumin is quenched strongly by the presence of core-shall QDs and follow the Stern-Volmer (S-V) relation, whereas the lifetime does not follow the S-V relation, demonstrating that the observed quenching is predominantly static in nature. Among all the QDs, the CdSe/ZnS QDs shows the least cytotoxicity hence most biocompatibility.

Keywords: biocompatibility, core-shell quantum dots, photoluminescence and lifetime, sensing ability

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506 Immunoglobulins and Importance in Ruminants

Authors: M. Akoz, O. B. Citil, I. Aydin

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Colostrum secreted by the mammary glands after birth in the early days, a high proportion of fat, protein and ash containing a secretion containing low amounts of casein and lactose. Especially immunoglobulins contain high proportions. Maternal immunoglobulins own immune system to protect the newborn against neonatal disease until development are very important matter. However, colostrum is transferred to the offspring due to placental barrier in ruminants. Immunoglobulins are absorbed through the intestinal epithelium but absorption can vary under the influence of some factors. These factors are among the priority ones taking colostrum first time, amount, concentration, the metabolic status of the newborn. intestinal absorption of immunoglobulins occurs over the first 24 h high. Absorption from the gut after nine hours, 50% after 24 hours was only 11%. On the other hand pup's digestive system degrade the enzymes after 24 hours immunoglobulins. Bovine colostrum in the composition while basic immune IgG, IgA and IgM are also available. Total IgG in colostrum of ruminants, while in other species is a greater amount in blood serum.

Keywords: immunoglobulin, ruminants, colostrum, immune system

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505 Prominent Lipid Parameters Correlated with Trunk-to-Leg and Appendicular Fat Ratios in Severe Pediatric Obesity

Authors: Mustafa M. Donma, Orkide Donma

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The examination of both serum lipid fractions and body’s lipid composition are quite informative during the evaluation of obesity stages. Within this context, alterations in lipid parameters are commonly observed. The variations in the fat distribution of the body are also noteworthy. Total cholesterol (TC), triglycerides (TRG), low density lipoprotein-cholesterol (LDL-C), high density lipoprotein-cholesterol (HDL-C) are considered as the basic lipid fractions. Fat deposited in trunk and extremities may give considerable amount of information and different messages during discrete health states. Ratios are also derived from distinct fat distribution in these areas. Trunk-to-leg fat ratio (TLFR) and trunk-to-appendicular fat ratio (TAFR) are the most recently introduced ratios. In this study, lipid fractions and TLFR, as well as TAFR, were evaluated, and the distinctions among healthy, obese (OB), and morbid obese (MO) groups were investigated. Three groups [normal body mass index (N-BMI), OB, MO] were constituted from a population aged 6 to 18 years. Ages and sexes of the groups were matched. The study protocol was approved by the Non-interventional Ethics Committee of Tekirdag Namik Kemal University. Written informed consent forms were obtained from the parents of the participants. Anthropometric measurements (height, weight, waist circumference, hip circumference, head circumference, neck circumference) were obtained and recorded during the physical examination. Body mass index values were calculated. Total, trunk, leg, and arm fat mass values were obtained by TANITA Bioelectrical Impedance Analysis. These values were used to calculate TLFR and TAFR. Systolic (SBP) and diastolic blood pressures (DBP) were measured. Routine biochemical tests including TC, TRG, LDL-C, HDL-C, and insulin were performed. Data were evaluated using SPSS software. p value smaller than 0.05 was accepted as statistically significant. There was no difference among the age values and gender ratios of the groups. Any statistically significant difference was not observed in terms of DBP, TLFR as well as serum lipid fractions. Higher SBP values were measured both in OB and MO children than those with N-BMI. TAFR showed a significant difference between N-BMI and OB groups. Statistically significant increases were detected between insulin values of N-BMI group and OB as well as MO groups. There were bivariate correlations between LDL and TLFR (r=0.396; p=0.037) as well as TAFR values (r=0.413; p=0.029) in MO group. When adjusted for SBP and DBP, partial correlations were calculated as (r=0.421; p=0.032) and (r=0.438; p=0.025) for LDL-TLFR as well as LDL-TAFR, respectively. Much stronger partial correlations were obtained for the same couples (r=0.475; p=0.019 and r=0.473; p=0.020, respectively) upon controlling for TRG and HDL-C. Much stronger partial correlations observed in MO children emphasize the potential transition from morbid obesity to metabolic syndrome. These findings have concluded that LDL-C may be suggested as a discriminating parameter between OB and MO children.

Keywords: children, lipid parameters, obesity, trunk-to-leg fat ratio, trunk-to-appendicular fat ratio

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504 Anti-DNA Antibodies from Patients with Schizophrenia Hydrolyze DNA

Authors: Evgeny A. Ermakov, Lyudmila P. Smirnova, Valentina N. Buneva

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Schizophrenia associated with dysregulation of neurotransmitter processes in the central nervous system and disturbances in the humoral immune system resulting in the formation of antibodies (Abs) to the various components of the nervous tissue. Abs to different neuronal receptors and DNA were detected in the blood of patients with schizophrenia. Abs hydrolyzing DNA were detected in pool of polyclonal autoantibodies in autoimmune and infectious diseases, such catalytic Abs were named abzymes. It is believed that DNA-hydrolyzing abzymes are cytotoxic, cause nuclear DNA fragmentation and induce cell death by apoptosis. Abzymes with DNAase activity are interesting because of the mechanism of formation and the possibility of use as diagnostic markers. Therefore, in our work we have set following goals: to determine the level anti-DNA Abs in the serum of patients with schizophrenia and to study DNA-hydrolyzing activity of IgG of patients with schizophrenia. Materials and methods: In our study there were included 41 patients with a verified diagnosis of paranoid or simple schizophrenia and 24 healthy donors. Electrophoretically and immunologically homogeneous IgGs were obtained by sequential affinity chromatography of the serum proteins on protein G-Sepharose and gel filtration. The levels of anti-DNA Abs were determined using ELISA. DNA-hydrolyzing activity was detected as the level of supercoiled pBluescript DNA transition in circular and linear forms, the hydrolysis products were analyzed by agarose electrophoresis followed by ethidium bromide stain. To correspond the registered catalytic activity directly to the antibodies we carried out a number of strict criteria: electrophoretic homogeneity of the antibodies, gel filtration (acid shock analysis) and in situ activity. Statistical analysis was performed in ‘Statistica 9.0’ using the non-parametric Mann-Whitney test. Results: The sera of approximately 30% of schizophrenia patients displayed a higher level of Abs interacting with single-stranded (ssDNA) and double-stranded DNA (dsDNA) compared with healthy donors. The average level of Abs interacting with ssDNA was only 1.1-fold lower than that for interacting with dsDNA. IgG of patient with schizophrenia were shown to possess DNA hydrolyzing activity. Using affinity chromatography, electrophoretic analysis of isolated IgG homogeneity, gel filtration in acid shock conditions and in situ DNAse activity analysis we proved that the observed activity is intrinsic property of studied antibodies. We have shown that the relative DNAase activity of IgG in patients with schizophrenia averaged 55.4±32.5%, IgG of healthy donors showed much lower activity (average of 9.1±6.5%). It should be noted that DNAase activity of IgG in patients with schizophrenia with a negative symptoms was significantly higher (73.3±23.8%), than in patients with positive symptoms (43.3±33.1%). Conclusion: Anti-DNA Abs of patients with schizophrenia not only bind DNA, but quite efficiently hydrolyze the substrate. The data show a correlation with the level of DNase activity and leading symptoms of patients with schizophrenia.

Keywords: anti-DNA antibodies, abzymes, DNA hydrolysis, schizophrenia

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503 Altered Gene Expression: Induction/Suppression of some Pathogenesis Related Protein Genes in an Egyptian Isolate of Potato Leafroll Virus (PLRV)

Authors: Dalia G. Aseel

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The potato (Solanum tubersum, L.) has become one of the major vegetable crops in Egypt and all over the world. Potato leafroll virus(PLRV) was observed on potato plants collected from different governorates in Egypt. Three cultivars, Spunta, Diamont, and Cara, infected with PLRV were collected; RNA was extracted and subjected to Real-Time PCR using the coat protein gene primers. The results showed that the expression of the coat protein was 39.6-fold, 12.45-fold, and 47.43-fold, respectively, for Spunta, Diamont, and Cara cultivars. Differential Display Polymerase Chain Reaction (DD-PCR) using pathogenesis-related protein 1 (PR-1), β-1,3-glucanases (PR-2), chitinase (PR-3), peroxidase (POD), and polyphenol oxidase (PPO) forward primers for pathogenesis-related proteins (PR). The obtained data revealed different banding patterns depending on the viral type and the region of infection. Regarding PLRV, 58 up-regulated and 19 down-regulated genes were detected. Sequence analysis of the up-and down-regulated genes revealed that infected plants were observed in comparison with the healthy control. Sequence analysis of the up-regulated gene was performed, and the encoding sequence analysis showed that the obtained genes include: induced stolen tip protein. On the other hand, two down-regulated genes were identified: disease resistance RPP-like protein and non-specific lipid-transfer protein. In this study, the expressions of PR-1, PR-2, PR-3, POD, and PPO genes in the infected leaves of three potato cultivars were estimated by quantitative real-time PCR. We can conclude that the PLRV-infection of potato plants inhibited the expression of the five PR genes. On the contrary, infected leaves by PLRV elevated the expression of some defense genes. This interaction may also induce and/or suppress the expression of some genes responsible for the plant's defense mechanisms.

Keywords: PLRV, pathogenesis-related proteins (PRs), DD-PCR, sequence, real-time PCR

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502 Assessment of Water Pollution in the River Nile (Egypt) by Applying Blood Biomarkers in Two Excellent Model Species Oreochromis niloticus niloticus and Clarias gariepinus

Authors: Alaa G. M. Osman, Abd-El –Baset M. Abd El Reheem, Khaled Y. Abouelfadl, Usama M. Mahmoud, Mohsen A. Moustafa

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This study aimed to explore new sites of biomarker research and to establish the use of blood parameters in wild fish populations. Four hundred and twenty fish samples were collected from six sites along the whole course of the river Nile, Egypt. The mean values of erythrocytes, thrombocytes, hemoglobin concentration, hematocrit value, and mean corpuscular volume were significantly lower in the blood of Nile tilapia and African catfish collected from downstream (contaminated) compared to upstream sites. In contrast, mean corpuscular hemoglobin and mean corpuscular hemoglobin concentration in the peripheral blood of both fish species significantly increased from upstream to downstream river Nile. The leukocytes count was significantly decreased in contaminated sites compared to upstream area. Hematological variables in the peripheral blood of Oreochromis niloticus niloticus and Clarias gariepinus exhibited significant (p<0.05) correlation with nearly all the detected chemical and physical parameters along the Nile course. In the present study, lower cellular and nuclear areas and cellular and nuclear shape factor were recorded in the erythrocytes of fish collected from downstream compared to those caught from upstream sites. This was confirmed by higher immature ratios of red cells in the blood of fish sampled from downstream river Nile. Karyorrhetic and enucleated erythrocytes were significantly correlated with physiochemical parameters in water samples collected from the same sites is being higher in the blood of fish collected from downstream sites. To see if there was any correlation between fish altered physiological fitness and environmental stress, we measured serum biochemical variables namely; total protein, cholesterol, triglycerides, calcium, chlorides, alkaline phosphatase activity (ALP), aspartate aminotransferase (AST), alanine aminotransferase (ALT), uric acid activity, creatinine, and serum glucose. The level of all the selected biochemical variables in the blood of O. niloticus niloticus and C. gariepinus were recorded to be significantly higher (p<0.05) in downstream sites. According to the present results, nearly all the detected haematological and blood biochemical variables are suitable indicators of contaminant exposure in O. niloticus niloticus and C. gariepinus. Also the detected erythrocytes malformations in blood collected from Nile tilapia and African catfish were proven to be suitable for bio-monitoring aquatic pollution. The results revealed species-specific differences in sensitivities, suggesting that Nile tilapia may serve as a more sensitive test species compared to African catfish.

Keywords: biomarkers, water pollution, blood parameters, river nile, african catfish, nile tilapia

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501 The Haemoglobin, Transferrin, Ceruloplasmin and Glutathione Polymorphism of Native Goat Breeds of Turkey, II-Kilis and Honamli

Authors: Ayse Ozge Demir, Nihat Mert

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In this research, Kilis and Honamli goats are used, which are specific local genetic resources of Turkey. The herds were independent, but they had similar care and nutrition circumstances. From each breed 30 samples were taken, in all 120 samples were collected. Erytrocyte, all blood and serum samples were used for hemoglobine (Hb), glutathione (GSH) and Tf with Cp analysis, respectively. In the analysis of this samples, Hb and Tf bands were determined by electrophoresis. However, Cp and GSH levels were analyzed by the spectrophotometer. Three Hb phenotypes (AA, BB, AB) and Six Tf phenotypes (AA, AB, AC, BB, BC, CC) were determined in this study. In addition, both the observed and the expected values of polymorphic characteristic for 2 characters were presented according to the Hardy-Weinberg Equilibrium (HWE). Cp levels were detected as 0.822 ± 0.055 mg/dl and 1.793 ± 0.109 mg/dl in Kilis and Honamli herds, respectively. GSH levels were detected as, 42,486 ± 1,034 mg/dl and 33.515 ± 0.345 mg/dl in these breeds, respectively,. On the other hand, the high and low GSH levels (GSHH and GSHh) of herds were presented.

Keywords: electrophoresis, gene resource, goat, spectrophotometer

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500 Prevalence of Positive Serology for Celiac Disease in Children With Autism Spectrum Disorder

Authors: A. Venkatakrishnan, M. Juneja, S. Kapoor

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Background: Gastrointestinal dysfunction is an emerging co morbidity seen in autism and may further strengthen the association between autism and celiac disease. This is supported by increased rates (22-70%) of gastrointestinal symptoms like diarrhea, constipation, abdominal discomfort/pain, and gastrointestinal inflammation in children with the etiology of autism is still elusive. In addition to genetic factors, environmental factors such as toxin exposure, intrauterine exposure to certain teratogenic drugs, are being proposed as possible contributing factors in the etiology of Autism Spectrum Disorders (ASD) in cognizance with reports of increased gut permeability and high rates of gastrointestinal symptoms noted in children with ASD, celiac disease has also been proposed as a possible etiological factor. Despite insufficient evidence regarding the benefit of restricted diets in Autism, GFD has been promoted as an alternative treatment for ASD. This study attempts to discern any correlation between ASD and celiac disease. Objective: This cross sectional study aims to determine the proportion of celiac disease in children with ASD. Methods: Study included 155 participants aged 2-12 yrs, diagnosed as ASD as per DSM-5 attending the child development center at a tertiary care hospital in Northern India. Those on gluten free diet or having other autoimmune conditions were excluded. A detailed Performa was filled which included sociodemographic details, history of gastrointestinal symptoms, anthropometry, systemic examination, and pertinent psychological testing was done using was assessed using Developmental Profile-3(DP-3) for Developmental Quotient, Childhood Autism Rating Scale-2 (CARS-2) for severity of ASD, Vineland Adaptive Behavior Scales (VABS) for adaptive behavior, Child Behavior Checklist (CBCL) for behavioral problems and BAMBI (Brief Autism Mealtime Behavior Scales) for feeding problems. Screening for celiac was done by TTG-IgA levels, and total serum IgA levels were measured to exclude IgA deficiency. Those with positive screen were further planned for HLA typing and endoscopic biopsy. Results: A total of 155 cases were included, out of which 5 had low IgA levels and were hence excluded from the study. The rest 150 children had TTG levels below the ULN and normal total serum IgA level. History of Gastrointestinal symptoms was present in 51 (34%) cases abdominal pain was the most frequent complaint (16.6%), followed by constipation (12.6%). Diarrhea was seen in 8 %. Gastrointestinal symptoms were significantly more common in children with ASD above 5 yrs (p-value 0.006) and those who were verbal (p = 0.000). There was no significant association between socio-demographic factors, anthropometric data, or severity of autism with gastrointestinal symptoms. Conclusion: None of the150 patients with ASD had raised TTG levels; hence no association was found between ASD and celiac disease. There is no justification for routine screening for celiac disease in children with ASD. Further studies are warranted to evaluate association of Non Celiac Gluten Sensitivity with ASD and any role of gluten-free diet in such patients.

Keywords: autism, celiac, gastrointestinal, gluten

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499 Green Synthesis of Red-Fluorescent Gold Nanoclusters: Characterization and Application for Breast Cancer Detection

Authors: Agnė Mikalauskaitė, Renata Karpicz, Vitalijus Karabanovas, Arūnas Jagminas

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The use of biocompatible precursors for the synthesis and stabilization of fluorescent gold nanoclusters (NCs) with strong red photoluminescence creates an important link between natural sciences and nanotechnology. Herein, we report the cost-effective synthesis of Au nanoclusters by templating and reduction of chloroauric acid with the cheap amino acid food supplements. This synthesis under the optimized conditions leads to the formation of biocompatible Au NCs having good stability and intense red photoluminescence, peaked at 680 to 705 nm, with a quantum yield (QY) of ≈7% and the average lifetime of up to several µs. The composition and luminescent properties of the obtained NCs were compared with ones formed via well-known bovine serum albumin reduction approach. Our findings implied that synthesized Au NCs tend to accumulate in more tumorigenic breast cancer cells (line MDA-MB-213) and after dialysis can be prospective for bio imagining.

Keywords: gold nanoclusters, proteins, materials chemistry, red-photoluminescence, bioimaging

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498 Preparation and Quality Control of a New Radiolabelled Complex of Spion

Authors: H. Yousefnia, SJ. Ahmadi, S. Sajadi, S. Zolghadri, A. Bahrami-Samani, M. Bagherzadeh

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Nowadays, superparamagnetic iron oxide nanoparticles (SPIONs) as the multitask agents have showed advantageous characteristics. The aim of this study was the preparation and quality control of 153Sm-DTPA-DA-SPION complex. Samarium-153 was produced by neutron irradiation of the enriched 152Sm2O3 in a research reactor for 5 d. For radiolabeling purposes, 8 mg of the ligand was added to the vial containing 153SmCl3 and the mixture was sonicated 30 min, while pH was adjusted to 7-8. The radiochemical purity of the complex was checked by the ITLC method using NH4OH:MeOH:H2O (0.2:2:4) as the mobile phase. This new radiolabeled complex was prepared with a radiochemical purity of higher than 98% in 30 min at the optimized condition. The complex was kept at room temperature and in human serum at 37 °C for 48 h, showed no loss of 153Sm from the complex. Considering all of these features, this new radiolabeled complex can be considered as a good therapeutic agent; however, further studies on its biological behavior are still needed.

Keywords: iron nanoparticles, preparation, quality control, 153Sm

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497 Association of Selected Biochemical Markers and Body Mass Index in Women with Endocrine Disorders

Authors: M. Mydlárová Blaščáková, J. Bernasovská, J. Poráčová, I. Boroňová

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Obesity is frequent attendant phenomenon of patients with endocrinological disease. Between BMI and endocrinological diseases is close correlation. In thesis we focused on the allocation of hormone concentration – PTH and TSH, CHOL a mineral element Ca in a blood serum. The examined group was formed by 100 respondents (women) aged 36 – 83 years, who were divided into two groups – control group (CG), group with diagnosed endocrine disease (DED). The concentration of PTH and TSH, Ca and CHOL was measured through the medium of analyzers Cobas e411 (Japan); Cobas Integra 400 (Switzerland). At individuals was measured body weight as well as stature and thereupon from those data we enumerated BMI. On the basis of Student T-test in biochemical parameter of PTH and Ca we found out significantly meaningful difference (p<0,05) between CG and DED. In CG we made a founding the association between BMI and PTH by means of correlation analysis.

Keywords: biochemical markers, hormones, obesity, women

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496 Active Exopolysaccharides Based Edible Coating Enriched with Red Seaweed (Gracilaria gracilis) Extract for Improved Preservation of Shrimp Quality during Refrigerated Storage

Authors: Rafik Balti, Mohamed Ben Mansour, Abdellah Arhaliass, Anthony Masse

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Unfortunately, shrimps are highly perishable and they start deteriorating immediately after death owing to their high water content and nutritional components. Currently, there has been an increasing interest in bioactive edible films and coatings to preserve the freshness and quality of foods. In this study, active edible coatings from microalgal exopolysaccharides (EPS) enriched with different concentrations of Red Seaweed Extract (RSE) (0.5, 1 and 1.5 % (w/v)) were developed and their effects on the quality changes of white shrimp during refrigerated storage (4 ± 1 °C) were examined over a period of 8 days. The control and the coated shrimp samples were analyzed periodically for microbiological (total viable bacteria, psychrotrophic bacteria, and enterobacteriaceae counts), chemical (pH, TVB-N, TMA-N, PV, TBARS), textural and sensory characteristics. The results indicated that the coating with a mixture of EPS and RSE could significantly decrease the total volatile basic nitrogen (TVB-N), trimethylamine (TMA) and thiobarbituric acid reactive substances (TBARS) (p < 0.05). With storage, EPS coatings containing RSE at both levels (1 and 1.5 %) were more effective in inhibiting the microbial species studied, specially psychrotrophic bacteria. Also, EPS + RSE coated samples had lower polyphenol oxidase (PPO) activity and lipid oxidation (p < 0.05) toward the end of storage. Textural and color properties of coated shrimp were generally more acceptable. Sensory scores indicated no significant changes in all samples during storage. The obtained results indicate that the edible EPS coating solutions enriched with RSE have noticeable effects on the quality and shelf life of shrimps when compared to control group. Finally, the present work demonstrates the effectiveness of EPS enriched coatings, offering a promising alternative to preserve more better the quality characteristics and to extend the shelf life of shrimp during the refrigerated storage

Keywords: active coating, exopolysaccharides, red seaweed, refrigerated storage, white shrimp

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495 Antiasthmatic Effect of Kankasava in OVA-Induced Asthma Mouse Model

Authors: Bharti Ahirwar

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The main object of this study was to evaluate the effect of kankasava on OVA-induced asthma in mouse model. Present study has demonstrated that kankasava exhibited an antiasthmatic effect by attenuated AHR and reducing level of IgE, IL-5, and IL-13, in both serum and BALF in OVA induced asthmatic mice. Effect of kankasav on airway responsiveness was obtained by monitoring the enhanced pen value . Kankasava significantly reduced AHR can be explained, in part, by reduction in both IgE overexoression and cytokine levels. Kankasava significantly decreased IL-4, IL-5, and IL-13 in BALF indicate that it may suppress the excess activity of T-cells and Th2 cytokines, which are implicated in the pathogenesis of allergic asthma, and consequently restore the Th1/Th2 imbalance of the immune system. In summary, we hypothesize that kankasava effectively suppressed elevations in IgE and cytokines levels, AHR, and mucus overproduction in mice with OVA-induced asthma suggested kankasava could be effective in immunological and pharmacological modulation of allergic asthma.

Keywords: asthma, ayurveda, kankasava, cytokine

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494 A Thermosensitive Polypeptide Hydrogel for Biomedical Application

Authors: Chih-Chi Cheng, Ji-Yu Lin, I-Ming Chu

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In this study, we synthesized a thermosensitive polypeptide hydrogel by copolymerizing poloxamer (PLX) and poly(ʟ-alanine) with ʟ-lysine segments at the both ends to form PLX-b-poly(ʟ-alanine-lysine) (Lys-Ala-PLX-Ala-Lys) copolymers. Poly(ʟ-alanine) is the hydrophobic chain of Lys-Ala-PLX-Ala-Lys copolymers which was designed to capture the hydrophobic agents. The synthesis was examined by 1H NMR and showed that Lys-Ala-PLX-Ala-Lys copolymers were successfully synthesized. At the concentration range of 3-7 wt%, the aqueous copolymer solution underwent sol-gel transition near the physiological temperature and exhibited changes in its secondary structure content, as evidenced by FTIR. The excellent viability of cells cultured within the scaffold was observed after 72 hr of incubation. Also, negatively charged bovine serum albumin was incorporated into the hydrogel without diminishing material integrity and shows good release profile. In the animal study, the results also indicated that Lys-Ala-PLX-Ala-Lys hydrogel has high potential in wound dressing.

Keywords: polypeptide thermosensitive hydrogel, tacrolimus, vascularized composite allotransplantation, sustain release

Procedia PDF Downloads 279
493 Effect of Formulated Insect Enriched Sprouted Soybean /Millet Based Food on Gut Health Markers in Albino Wistar Rats

Authors: Gadanya, A.M., Ponfa, S., Jibril, M.M., Abubakar, S. M.

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Background: Edible insects such as grasshopper are important sources of food for humans, and have been consumed as traditional foods by many indigenous communities especially in Africa, Asia, and Latin America. These communities have developed their skills and techniques in harvesting, preparing, consuming, and preserving edible insects, widely contributing to the role played by the use of insects in human nutrition. Aim/ objective: This study was aimed at determining the effect of insect enriched sprouted soyabean /millet based food on some gut health markers in albino rats. Methods. Four different formulations of Complementary foods (i.e Complementary Food B (CFB): sprouted millet (SM), Complementary Food C (CFC): sprouted soyabean (SSB), Complementary Food D (CFD): sprouted soybean and millet (SSBM) in a ratio of (50:50) and Complementary Food E (CFE): insect (grasshopper) enriched sprouted soybean and millet (SSBMI) in a ratio of (50:25:25)) were prepared. Proximate composition and short chain fatty acid contents were determined. Thirty albino rats were divided into5 groups of six rats each. Group 1(CDA) were fed with basal diet and served as a control group, while groups 2,3,4 and 5 were fed with the corresponding complimentary foods CFB, CFC, CFD and CFE respectively daily for four weeks. Concentrations of fecal protein, serum total carotenoids and nitric oxide were determined. DNA extraction for molecular isolation and characterization were carried out followed by PCR, the use of mega 11 software and NCBI blast for construction of the phylogenetic tree and organism identification respectively. Results: Significant increase (P<0.05) in percentage ash, fat, protein and moisture contents, as well as short chain fatty acid (acetate, butyrate and propionate) concentrations were recorded in the insect enriched sprouted composite food (CFE) when compared with the CFA, CFB, CFC and CFD composite food. Faecal protein, carotenoid and nitric oxide concentrations were significantly lower (P>0.05) in group 5 in comparison to groups 1to 4. Ruminococcus bromii and Bacteroidetes were molecularly isolated and characterized by 16s rRNA from the sprouted millet/sprouted soybean and the insect enriched sprouted soybean/sprouted millet based food respectively. The presence of these bacterial strains in the feaces of the treated rats is an indication that the gut of the treated rats is colonized by good gut bacteria, hence, an improved gut health. Conclusion: Insect enriched sprouted soya bean/sprouted millet based complementary diet showed a high composition of ash, fat, protein and fiber. Thus, could increase the availability of short chain fatty acids whose role to the host organism cannot be overemphasized. It was also found to have decrease the level of faecal protein, carotenoid and nitric oxide in the serum which is an indication of an improvement in the immune system function.

Keywords: gut-health, insect, millet, soybean, sprouted

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492 SEC-MALLS Study of Hyaluronic Acid and BSA Thermal Degradation in Powder and in Solution

Authors: Vasile Simulescu, Jakub Mondek, Miloslav Pekař

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Hyaluronic acid (HA) is an anionic glycosaminoglycan distributed throughout connective, epithelial and neural tissues. The importance of hyaluronic acid increased in the last decades. It has many applications in medicine and cosmetics. Hyaluronic acid has been used in attempts to treat osteoarthritis of the knee via injecting it into the joint. Bovine serum albumin (also known as BSA) is a protein derived from cows, which has many biochemical applications. The aim of our research work was to compare the thermal degradation of hyaluronic acid and BSA in powder and in solution, by determining changes in molar mass and conformation, by using SEC-MALLS (size exclusion chromatography -multi angle laser light scattering). The aim of our research work was to observe the degradation in powder and in solution of different molar mass hyaluronic acid samples, at different temperatures for certain periods. The degradation of the analyzed samples was mainly observed by modifications in molar mass.

Keywords: thermal degradation, hyaluronic acid, BSA, SEC-MALLS

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491 Traumatic Brain Injury Induced Lipid Profiling of Lipids in Mice Serum Using UHPLC-Q-TOF-MS

Authors: Seema Dhariwal, Kiran Maan, Ruchi Baghel, Apoorva Sharma, Poonam Rana

Abstract:

Introduction: Traumatic brain injury (TBI) is defined as the temporary or permanent alteration in brain function and pathology caused by an external mechanical force. It represents the leading cause of mortality and morbidity among children and youth individuals. Various models of TBI in rodents have been developed in the laboratory to mimic the scenario of injury. Blast overpressure injury is common among civilians and military personnel, followed by accidents or explosive devices. In addition to this, the lateral Controlled cortical impact (CCI) model mimics the blunt, penetrating injury. Method: In the present study, we have developed two different mild TBI models using blast and CCI injury. In the blast model, helium gas was used to create an overpressure of 130 kPa (±5) via a shock tube, and CCI injury was induced with an impact depth of 1.5mm to create diffusive and focal injury, respectively. C57BL/6J male mice (10-12 weeks) were divided into three groups: (1) control, (2) Blast treated, (3) CCI treated, and were exposed to different injury models. Serum was collected on Day1 and day7, followed by biphasic extraction using MTBE/Methanol/Water. Prepared samples were separated on Charged Surface Hybrid (CSH) C18 column and acquired on UHPLC-Q-TOF-MS using ESI probe with inhouse optimized parameters and method. MS peak list was generated using Markerview TM. Data were normalized, Pareto-scaled, and log-transformed, followed by multivariate and univariate analysis in metaboanalyst. Result and discussion: Untargeted profiling of lipids generated extensive data features, which were annotated through LIPID MAPS® based on their m/z and were further confirmed based on their fragment pattern by LipidBlast. There is the final annotation of 269 features in the positive and 182 features in the negative mode of ionization. PCA and PLS-DA score plots showed clear segregation of injury groups to controls. Among various lipids in mild blast and CCI, five lipids (Glycerophospholipids {PC 30:2, PE O-33:3, PG 28:3;O3 and PS 36:1 } and fatty acyl { FA 21:3;O2}) were significantly altered in both injury groups at Day 1 and Day 7, and also had VIP score >1. Pathway analysis by Biopan has also shown hampered synthesis of Glycerolipids and Glycerophospholipiods, which coincides with earlier reports. It could be a direct result of alteration in the Acetylcholine signaling pathway in response to TBI. Understanding the role of a specific class of lipid metabolism, regulation and transport could be beneficial to TBI research since it could provide new targets and determine the best therapeutic intervention. This study demonstrates the potential lipid biomarkers which can be used for injury severity diagnosis and identification irrespective of injury type (diffusive or focal).

Keywords: LipidBlast, lipidomic biomarker, LIPID MAPS®, TBI

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490 Tailorability of Poly(Aspartic Acid)/BSA Complex by Self-Assembling in Aqueous Solutions

Authors: Loredana E. Nita, Aurica P. Chiriac, Elena Stoleru, Alina Diaconu, Tudorachi Nita

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Self-assembly processes are an attractive method to form new and complex structures between macromolecular compounds to be used for specific applications. In this context, intramolecular and intermolecular bonds play a key role during self-assembling processes in preparation of carrier systems of bioactive substances. Polyelectrolyte complexes (PECs) are formed through electrostatic interactions, and though they are significantly below of the covalent linkages in their strength, these complexes are sufficiently stable owing to the association processes. The relative ease way of PECs formation makes from them a versatile tool for preparation of various materials, with properties that can be tuned by adjusting several parameters, such as the chemical composition and structure of polyelectrolytes, pH and ionic strength of solutions, temperature and post-treatment procedures. For example, protein-polyelectrolyte complexes (PPCs) are playing an important role in various chemical and biological processes, such as protein separation, enzyme stabilization and polymer drug delivery systems. The present investigation is focused on evaluation of the PPC formation between a synthetic polypeptide (poly(aspartic acid) – PAS) and a natural protein (bovine serum albumin - BSA). The PPC obtained from PAS and BSA in different ratio was investigated by corroboration of various techniques of characterization as: spectroscopy, microscopy, thermo-gravimetric analysis, DLS and zeta potential determination, measurements which were performed in static and/or dynamic conditions. The static contact angle of the sample films was also determined in order to evaluate the changes brought upon surface free energy of the prepared PPCs in interdependence with the complexes composition. The evolution of hydrodynamic diameter and zeta potential of the PPC, recorded in situ, confirm changes of both co-partners conformation, a 1/1 ratio between protein and polyelectrolyte being benefit for the preparation of a stable PPC. Also, the study evidenced the dependence of PPC formation on the temperature of preparation. Thus, at low temperatures the PPC is formed with compact structure, small dimension and hydrodynamic diameter, close to those of BSA. The behavior at thermal treatment of the prepared PPCs is in agreement with the composition of the complexes. From the contact angle determination results the increase of the PPC films cohesion, which is higher than that of BSA films. Also, a higher hydrophobicity corresponds to the new PPC films denoting a good adhesion of the red blood cells onto the surface of PSA/BSA interpenetrated systems. The SEM investigation evidenced as well the specific internal structure of PPC concretized in phases with different size and shape in interdependence with the interpolymer mixture composition.

Keywords: polyelectrolyte – protein complex, bovine serum albumin, poly(aspartic acid), self-assembly

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