Search results for: serum protein electrophoresis
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 3135

Search results for: serum protein electrophoresis

585 Investigation on Single Nucleotide Polymorphism in Candidate Genes and Their Association with Occurrence of Mycobacterium avium Subspecies Paratuberculosis Infection in Cattle

Authors: Ran Vir Singh, Anuj Chauhan, Subhodh Kumar, Rajesh Rathore, Satish Kumar, B Gopi, Sushil Kumar, Tarun Kumar, Ramji Yadav, Donna Phangchopi, Shoor Vir Singh

Abstract:

Paratuberculosis caused by Mycobacterium avium subspecies paratuberculosis (MAP) is a chronic granulomatous enteritis affecting ruminants. It is responsible for significant economic losses in livestock industry worldwide. This organism is also of public health concern due to an unconfirmed link to Crohn’s disease. Susceptibility to paratuberculosis has been suggested to have genetic component with low to moderate heritability. Number of SNPs in various candidates genes have been observed to be affecting the susceptibility toward paratuberculosis. The objective of this study was to explore the association of various SNPs in the candidate genes and QTL region with MAP. A total of 117 SNPs from SLC11A1, IFNG, CARD15, TLR2, TLR4, CLEC7A, CD209, SP110, ANKARA2, PGLYRP1 and one QTL were selected for study. A total of 1222 cattle from various organized herds, gauhsalas and farmer herds were screened for MAP infection by Johnin intradermal skin test, AGID, serum ELISA, fecal microscopy, fecal culture and IS900 blood PCR. Based on the results of these tests, a case and control population of 200 and 183 respectively was established for study. A total of 117 SNPs from 10 candidate genes and one QTL were selected and validated/tested in our case and control population by PCR-RFLP technique. Data was analyzed using SAS 9.3 software. Statistical analysis revealed that, 107 out of 117 SNPs were not significantly associated with occurrence of MAP. Only SNP rs55617172 of TLR2, rs8193046 and rs8193060 of TLR4, rs110353594 and rs41654445 of CLEC7A, rs208814257of CD209, rs41933863 of ANKRA2, two loci {SLC11A1(53C/G)} and {IFNG (185 G/r) } and SNP rs41945014 in QTL region was significantly associated with MAP. Six SNP from 10 significant SNPs viz., rs110353594 and rs41654445 from CLEC7A, rs8193046 and rs8193060 from TLR4, rs109453173 from SLC11A1 rs208814257 from CD209 were validated in new case and control population. Out of these only one SNP rs8193046 of TLR4 gene was found significantly associated with occurrence of MAP in cattle. ODD ratio indicates that animals with AG genotype were more susceptible to MAP and this finding is in accordance with the earlier report. Hence it reaffirms that AG genotype can serve as a reliable genetic marker for indentifying more susceptible cattle in future selection against MAP infection in cattle.

Keywords: SNP, candidate genes, paratuberculosis, cattle

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584 Lactobacillus rhamnosus GG Increases the Re-Epithelialization Rate of Model Wounds by Stimulating Keratinocyte Migration in Ex-Vivo

Authors: W. Mohammedsaeed, A. J. Mcbain, S. M. Cruickshank, C. A. O’Neill

Abstract:

Many studies have demonstrated the importance of probiotics and their potential therapeutic effects within the gut. Recently, the possible therapeutic effects of probiotics in other tissues have also begun to be investigated. Comparatively few studies have evaluated the use of topical probiotics in relation to the skin. In this study, we have conducted preliminary investigations into whether a well-known probiotic, Lactobacillus rhamnosus GG (LGG), can increase the rate of re-epithelialization in a model wound. Full-thickness skin was obtained from individuals undergoing elective cosmetic surgery. This skin was wounded using excisional punch and cultured using a serum-free medium, either in the presence or absence of L. rhamnosus GG lysate. Histological staining of the sections was performed with Haematoxylin& Eosin E to quantify “epithelial tongue length”. This is the length of the new epithelial ‘tongue’ that grows and covers the exposed dermis at the inner wound edges. The length of the new epithelial ‘tongue’ was compared in untreated section and section treated with and L. rhamnosus GG made using108CFU/ml bacterial cells. L. rhamnosus GG lysate enhanced significantly the re-epithelialisation of treated wounds compared with that of untreated wounds (P=0.005, n=3). Tongue length, at day 1 was 7.55μm 0.15, at day 3 it was 18.5μm 0.25 and at day 7 was 22.9μm 0.35. These results can be compared with untreated cultures in which tongue length was 3.25μm 0.35, day 3 was 9.65μm 0.25 and day 7 was 13.5μm 0.15 post-wounding. In ex-vivo proliferation and migration cells were measured by determining the expression of nuclear proliferation marker Ki-67 and the expression of Phosphorylated cortactin respectively demonstrated that L. rhamnosus GG significantly increased NHEK proliferation and migration rates relative to controls. However, the dominant mechanism was migration because in ex-vivo skin treated with the L. rhamnosus GG up-regulated the gene expression of the chemokine receptor and ligands CXCR2 and CXCL2 comparing with controls (P=0.02, P=0.03 respectively, n=3). High levels of CXCL2/CXCL2 have already been implicated in multiple aspects of stimulation of wound healing through activation of keratinocyte migration. These data demonstrate that lysates from Lactobacillus rhamnosus GG increase re-epithelialization by stimulation of keratinocyte migration. The current study identifies the partial mechanism that contribute to stimulating the wound-healing process ex vivo in response to L. rhamnosus GG lysate is an increase in the production of CXCL2/ CXCR2 in ex vivo models. The use of probiotic lysates potentially offers new options to develop treatments that could improve wound healing.

Keywords: Lactobacillus rhamnosus GG, wounds, migration, lysate

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583 Further Evidence for the Existence of Broiler Chicken PFN (Pale, Firm and Non-Exudative Meat) and PSE (Pale, Soft and Exudative) in Brazilian Commercial Flocks

Authors: Leila M. Carvalho, Maria Erica S. Oliveira, Arnoud C. Neto, Elza I. Ida, Massami Shimokomaki, Marta S. Madruga

Abstract:

The quality of broiler breast meat is changing as a result of the continuing emphasis on genetic selection for a more efficient meat production. Breast meat has been classified as PSE (pale, soft, exudative), DFD (dark, firm, dry) and normal color meat, and recently a third group has emerged: the so-called PFN (pale, firm, non-exudative) meat. This classification was based on pH, color and functional properties. The aim of this work was to confirm the existence of PFN and PSE meat by biochemical characterization and functional properties. Twenty four hours of refrigerated fillet, Pectoralis major, m. samples (n= 838) were taken from Cobb flocks 42-48 days old, obtained in Northeastern Brazil tropical region, the Northeastern, considered to have only dry and wet seasons. Color (L*), pH, water holding capacity (WHC), values were evaluated and compared with PSE group samples. These samples were classified as Normal (465.8), PSE meat (L*≥53; pH<5.8) and PFN (L*≥53; pH>5.8). The occurrence of control meat, PSE and PFN was 69.09%, 11.10% and 19.81%, respectively. Samples from PFN presented 4.0-5.0% higher WHC in relation to PSE meat and similar to control group. These results are explained by the fact that PSE meat syndrome occurs because of higher protein denaturation as the consequence of a simultaneous lower pH values under warm carcass sooner after slaughtering impairing the myofibril proteins functional properties. Conversely, PFN samples follow normal glycolysis rate maintaining the normal proteins activities. In conclusion, the results reported herein confirm the existence of this emerging broiler meat group with similar properties as control group and it should be considered as normal breast meat group.

Keywords: broiler breast meat, funcional properties, PFN, PSE

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582 Aorta Adhesion Molecules in Cholesterol-Fed Rats Supplemented with Extra Virgin Olive Oil or Sunflower Oil, in Either Commercial or Modified Forms

Authors: Ageliki I. Katsarou, Andriana C. Kaliora, Antonia Chiou, Apostolos Papalois, Nick Kalogeropoulos, Nikolaos K. Andrikopoulos

Abstract:

Chronic inflammation plays a pivotal role in CVD development, while phytochemicals have been shown to reduce CVD risk. Several studies have correlated olive oil consumption with CVD prevention and CVD risk reduction. However, the effect of individual olive oil macro- or micro-constituents and possible synergisms among them needs to be further elucidated. Herein, extra virgin olive oil (EVOO) lipidic and polar phenolics fractions were evaluated for their effect on inflammatory markers in cholesterol-fed rats. Oils combining different characteristics as to their polar phenolic content and lipid profile were used. Male Wistar rats were fed for 9 weeks on either a high-cholesterol diet (HCD) or a HCD supplemented with oils, either commercially available, i.e. EVOO, sunflower oil (SO), or modified as to their polar phenol content, i.e. phenolics deprived-EVOO (EVOOd), SO enriched with the EVOO phenolics (SOe). Post-intervention, aorta and blood samples were collected. HCD induced dyslipidemia, manifested by serum total cholesterol and low-density lipoprotein cholesterol elevation. Additionally, HCD resulted in higher adhesion molecules’ levels in rat aorta. In the case of E-selectin, this increase was attenuated by HCD supplementation with EVOO and EVOOd, while no alterations were observed in SO and SOe groups. No differences were observed between pairs of commercial and modified oils, indicating that oleates may be the components responsible for aorta E-selectin levels lowering. The same was true for vascular adhesion molecule-1 (VCAM-1); augmentation in cholesterol-fed animals was attenuated by EVOO and EVOOd diets, highlighting oleates effect. In addition, VCAM-1 levels were higher in SO group compared to the respective SOe, indicating that in the presence of phenolic compounds linoleic acid have become less prone to oxidation. Intercellular adhesion molecule-1 (ICAM-1) levels were higher in cholesterol-fed rats, however not affected by any of the oils supplemented during the intervention. Overall, EVOO was found superior in regulating adhesion molecule levels in rat aorta compared to SO. EVOO and EVOOd exhibited analogous effects on all adhesion molecules assessed, indicating that EVOO major constituents (oleates) improve E-selectin and VCAM-1 levels in rat aorta, independently from phenolics presence. Further research is needed to elucidate the effect of phenolics and oleates in other tissues.

Keywords: extra virgin olive oil, cholesterol-fed rats, polar phenolics, adhesion molecules

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581 Structural Characterization of TIR Domains Interaction

Authors: Sara Przetocka, Krzysztof Żak, Grzegorz Dubin, Tadeusz Holak

Abstract:

Toll-like receptors (TLRs) play central role in the innate immune response and inflammation by recognizing pathogen-associated molecular patterns (PAMPs). A fundamental basis of TLR signalling is dependent upon the recruitment and association of adaptor molecules that contain the structurally conserved Toll/interleukin-1 receptor (TIR) domain. MyD88 (myeloid differentiation primary response gene 88) is the universal adaptor for TLRs and cooperates with Mal (MyD88 adapter-like protein, also known as TIRAP) in TLR4 response which is predominantly used in inflammation, host defence and carcinogenesis. Up to date two possible models of MyD88, Mal and TLR4 interactions have been proposed. The aim of our studies is to confirm or abolish presented models and accomplish the full structural characterisation of TIR domains interaction. Using molecular cloning methods we obtained several construct of MyD88 and Mal TIR domain with GST or 6xHis tag. Gel filtration method as well as pull-down analysis confirmed that recombinant TIR domains from MyD88 and Mal are binding in complexes. To examine whether obtained complexes are homo- or heterodimers we carried out cross-linking reaction of TIR domains with BS3 compound combined with mass spectrometry. To investigate which amino acid residues are involved in this interaction the NMR titration experiments were performed. 15N MyD88-TIR solution was complemented with non-labelled Mal-TIR. The results undoubtedly indicate that MyD88-TIR interact with Mal-TIR. Moreover 2D spectra demonstrated that simultaneously Mal-TIR self-dimerization occurs which is necessary to create proper scaffold for Mal-TIR and MyD88-TIR interaction. Final step of this study will be crystallization of MyD88 and Mal TIR domains complex. This crystal structure and characterisation of its interface will have an impact in understanding the TLR signalling pathway and possibly will be used in development of new anti-cancer treatment.

Keywords: cancer, MyD88, TIR domains, Toll-like receptors

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580 EDTA Enhanced Plant Growth, Antioxidant Defense System, and Phytoextraction of Copper by Brassica napus L.

Authors: Ume Habiba, Shafaqat Ali, Mujahid Farid, Muhammad Bilal Shakoor

Abstract:

Copper (Cu) is an essential micronutrient for normal plant growth and development, but in excess, it is also toxic to plants. The present study investigated the influence of ethylenediaminetetraacetic acid (EDTA) in enhancing Cu uptake and tolerance as well as the morphological and physiological responses of Brassica napus L. seedlings under Cu stress. Four-week-old seedlings were transferred to hydroponics containing Hoagland’s nutrient solution. After 2 weeks of transplanting, three levels (0, 50, and 100 μM) of Cu were applied with or without application of 2.5 mM EDTA and plants were further grown for 8 weeks in culture media. Results showed that Cu alone significantly decreased plant growth, biomass, photosynthetic pigments, and gas exchange characteristics. Cu stress also reduced the activities of antioxidants, such as superoxide dismutase (SOD), peroxidase (POD), ascorbate peroxidase (APX), and catalase (CAT) along with protein contents. Cu toxicity increased the concentration of reactive oxygen species (ROS) as indicated by the increased production of malondialdehyde (MDA) and hydrogen peroxide (H2O2) in both leaves and roots. The application of EDTA significantly alleviated Cu-induced toxic effects in B. napus, showing remarkable improvement in all these parameters. EDTA amendment increased the activity of antioxidant enzymes by decreasing the concentrations of MDA and H2O2 both in leaves and roots of B. napus. Although, EDTA amendment with Cu significantly increased Cu uptake in roots, stems, and leaves in decreasing order of concentration but increased the growth, photosynthetic parameters, and antioxidant enzymes. These results showed that the application of EDTA can be a useful strategy for phytoextraction of Cu by B. napus from contaminated soils.

Keywords: antioxidants, biomass, copper, EDTA, phytoextraction, tolerance

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579 Exploring 1,2,4-Triazine-3(2H)-One Derivatives as Anticancer Agents for Breast Cancer: A QSAR, Molecular Docking, ADMET, and Molecular Dynamics

Authors: Said Belaaouad

Abstract:

This study aimed to explore the quantitative structure-activity relationship (QSAR) of 1,2,4-Triazine-3(2H)-one derivative as a potential anticancer agent against breast cancer. The electronic descriptors were obtained using the Density Functional Theory (DFT) method, and a multiple linear regression techniques was employed to construct the QSAR model. The model exhibited favorable statistical parameters, including R2=0.849, R2adj=0.656, MSE=0.056, R2test=0.710, and Q2cv=0.542, indicating its reliability. Among the descriptors analyzed, absolute electronegativity (χ), total energy (TE), number of hydrogen bond donors (NHD), water solubility (LogS), and shape coefficient (I) were identified as influential factors. Furthermore, leveraging the validated QSAR model, new derivatives of 1,2,4-Triazine-3(2H)-one were designed, and their activity and pharmacokinetic properties were estimated. Subsequently, molecular docking (MD) and molecular dynamics (MD) simulations were employed to assess the binding affinity of the designed molecules. The Tubulin colchicine binding site, which plays a crucial role in cancer treatment, was chosen as the target protein. Through the simulation trajectory spanning 100 ns, the binding affinity was calculated using the MMPBSA script. As a result, fourteen novel Tubulin-colchicine inhibitors with promising pharmacokinetic characteristics were identified. Overall, this study provides valuable insights into the QSAR of 1,2,4-Triazine-3(2H)-one derivative as potential anticancer agent, along with the design of new compounds and their assessment through molecular docking and dynamics simulations targeting the Tubulin-colchicine binding site.

Keywords: QSAR, molecular docking, ADMET, 1, 2, 4-triazin-3(2H)-ones, breast cancer, anticancer, molecular dynamic simulations, MMPBSA calculation

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578 Surface Modified Polyamidoamine Dendrimer with Gallic Acid Overcomes Drug Resistance in Colon Cancer Cells HCT-116

Authors: Khushbu Priyadarshi, Chandramani Pathak

Abstract:

Cancer cells can develop resistance to conventional therapies especially chemotherapeutic drugs. Resistance to chemotherapy is another challenge in cancer therapeutics. Therefore, it is important to address this issue. Gallic acid (GA) is a natural plant compound that exhibits various biological properties including anti-proliferative, anti-inflammatory, anti-oxidant and anti-bacterial. Despite of the wide spectrum biological properties GA has cytotoxic response and low bioavailability. To overcome this problem, GA was conjugated with the Polyamidoamine(PAMAM) dendrimer for improving the bioavailability and efficient delivery in drug-resistant HCT-116 Colon Cancer cells. Gallic acid was covalently linked to 4.0 G PAMAM dendrimer. PAMAM dendrimer is well established nanocarrier but has cytotoxicity due to presence of amphiphilic nature of amino group. In our study we have modified surface of PAMAM dendrimer with Gallic acid and examine their anti-proliferative effects in drug-resistant HCT-116 cells. Further, drug-resistant colon cancer cells were established and thereafter treated with different concentration of PAMAM-GA to examine their anti-proliferative potential. Our results show that PAMAM-GA conjugate induces apoptotic cell death in HCT-116 and drug-resistant cells observed by Annexin-PI staining. In addition, it also shows that multidrug-resistant drug transporter P-gp protein expression was downregulated with increasing the concentration of GA conjugate. After that we also observed the significant difference in Rh123 efflux and accumulation in drug sensitive and drug-resistant cancer cells. Thus, our study suggests that conjugation of anti-cancer agents with PAMAM could improve drug resistant property and cytotoxic response to treatment of cancer.

Keywords: drug resistance, gallic acid, PAMAM dendrimer, P-glycoprotein

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577 Method Development for the Determination of Gamma-Aminobutyric Acid in Rice Products by Lc-Ms-Ms

Authors: Cher Rong Matthew Kong, Edmund Tian, Seng Poon Ong, Chee Sian Gan

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Gamma-aminobutyric acid (GABA) is a non-protein amino acid that is a functional constituent of certain rice varieties. When consumed, it decreases blood pressure and reduces the risk of hypertension-related diseases. This has led to more research dedicated towards the development of functional food products (e.g. germinated brown rice) with enhanced GABA content, and the development of these functional food products has led to increased demand for instrument-based methods that can efficiently and effectively determine GABA content. Current analytical methods require analyte derivatisation, and have significant disadvantages such as being labour intensive and time-consuming, and being subject to analyte loss due to the increased complexity of the sample preparation process. To address this, an LC-MS-MS method for the determination of GABA in rice products has been developed and validated. This developed method involves a relatively simple sample preparation process before analysis using HILIC LC-MS-MS. This method eliminates the need for derivatisation, thereby significantly reducing the labour and time associated with such an analysis. Using LC-MS-MS also allows for better differentiation of GABA from any potential co-eluting compounds in the sample matrix. Results obtained from the developed method demonstrated high linearity, accuracy, and precision for the determination of GABA (1ng/L to 8ng/L) in a variety of brown rice products. The method can significantly simplify sample preparation steps, improve the accuracy of quantitation, and increase the throughput of analyses, thereby providing a quick but effective alternative to established instrumental analysis methods for GABA in rice.

Keywords: functional food, gamma-aminobutyric acid, germinated brown rice, method development

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576 The Effect of Four-Week Resistance Exercise along with Milk Consumption on NT-proBNP and Plasma Troponin I

Authors: Rostam Abdi, Ahmad Abdi, Zahra Vahedi Langrodi

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The aim of this study is to investigate four-week resistance exercise and milk supplement on NT-proBNP and plasma troponin I of male students. Concerning the methodology of the study, 21 senior high school students of Ardebil city were selected. The selected subjects were randomly shared in three groups of control, exercise- water and exercise- milk. The exercise program includes resistance exercise for a big muscle group. The subjects of control group rested during the study and did not participate in any training. The subjects of exercise- water experimental group immediately received 400 cc water after exercise and exercise- milk group immediately received 400 cc low fat milk. Control-water groups consumed the same amount of water. 48 hours before and after the last exercise session, the blood sample of the subjects were taken for measuring the variables. NT-proBNP and Troponin I concentrations were measured by ELISA. For data analysis, one-way variance analysis test, correlated t-test and Bonferroni post hoc test were used. The significant difference of p ≤ 0.05 was accepted. Resistance training along with milk consumption leads to increase of plasma NT-proBNP, however; this increase has not reached the significant level. Furthermore, meaningful increase was observed in plasma NT–proBNP in exercise group between pretest and posttest values. Furthermore, no meaningful difference was observed between groups in terms of Troponin I after milk consumption. It seems that endurance exercises lead to change in the structure of heart muscle and is along with an increase of NT-proBNP. Furthermore, there is the possibility that milk consumption can lead to release of heart troponin I. The mechanism through which protein supplements have been put on heart troponin I is unknown and requires more research.

Keywords: resistance exercise, milk, NT-proBNP, Troponin I

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575 The Impact of Diesel Exhaust Particles on Tight Junction Proteins on Nose and Lung in a Mouse Model

Authors: Kim Byeong-Gon, Lee Pureun-Haneul, Hong Jisu, Jang An-Soo

Abstract:

Background: Diesel exhaust particles (DEPs) lead to trigger airway hyperresponsiveness (AHR) and airway dysfunction or inflammation in respiratory systems. Whether tight junction protein changes can contribute to development or exacerbations of airway diseases remain to be clarified. Objective: The aim of this study was to observe the effect of DEP on tight junction proteins in one airway both nose and lung in a mouse model. Methods: Mice were treated with saline (Sham) and exposed to 100 μg/m³ DEPs 1 hour a day for 5 days a week for 4 weeks and 8 weeks in a closed-system chamber attached to a ultrasonic nebulizer. Airway hyperresponsiveness (AHR) was measured and bronchoalveolar lavage (BAL) fluid, nasal lavage (NAL) fluid, lung and nasal tissue was collected. The effects of DEP on tight junction proteins were estimated using western blot, immunohistochemical in lung and nasal tissue. Results: Airway hyperresponsiveness and number of inflammatory cells were higher in DEP exposure group than in control group, and were higher in 4 and 8 weeks model than in control group. The expression of tight junction proteins CLND4, -5, and -17 in both lung and nasal tissue were significantly increased in DEP exposure group than in the control group. Conclusion: These results suggesting that CLDN4, -5 and -17 may be involved in the airway both nose and lung, suggesting that air pollutants cause to disruption of epithelial and endothelial cell barriers. Acknowledgment: This research was supported by Korea Ministry of Environment (MOE) as 'The Environmental Health Action Program' (2016001360009) and Soonchunhyang University Research Fund.

Keywords: diesel exhaust particles, air pollutant, tight junction, Claudin, Airway inflammation

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574 Use of Recycled Vegetable Oil in the Diet of Lactating Sows

Authors: Juan Manuel Uriarte Lopez, Hector Raul Guemez Gaxiola, Javier Alonso Romo Rubio, Juan Manuel Romo Valdez

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The objective of this investigation was to determine the influence of the use of recycled vegetable oil from restaurants in the productive performance of sows in lactation. Twenty-four hybrids lactating sows (Landrace x Yorkshire) were divided into three treatments with eight sows per treatment. On day 107 of gestation, the sows were moved to the mesh floor maternity cages in an environment regulated by the environment regulated (2.4 × 0.6 m) contained an area (2.4 × 0.5 m) for newborn pigs on each side, all diets were provided as a dry powder, and the sows received free access to water throughout the experimental period. After farrowing, the sows were fasted for 12 hours, the daily feed ration gradually increased, and the sows had ad libitum access to feed on the fourth day. The diets used were corn-soybean meal-based, containing 0 (CONT), recycled vegetable oil 1.0 % (RVOL), or recycled vegetable oil 1.5 % (RVOH) for 30 days. The diets contained similar calculated levels of crude protein and metabolizable energy and contained vitamins and minerals that exceeded National Research Council (1998) recommendations; sows were fed three times daily. On day 30, piglets were weaned, and performances of lactating sows and nursery piglets were recorded. Results indicated that average daily feed intake (5.58, 5.55, and 5.49 kg for CONT, RVOL, and RVO, respectively) of sows were not affected (P > 0.05) by different dietary. There was no difference in the average body weight of piglets on the day of birth, with 1.33, 1.36, and 1.35 kg, respectively (P > 0.05). There was no difference in average body weight of piglets on day 30, with 6.91, 6.75, and 7.05 kg, respectively 0.05) between treatments numbers of weaned piglets per sow (9.95, 9.80, and 9.80) were not affected by treatments (P > 0.05).In conclusion, the substitution of virgin vegetable oil for recycled vegetable oil in the diet does not affect the productive performance of lactating sows.

Keywords: lactating, sow, vegetable, oil

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573 Molecular Mechanism on Inflammation and Antioxidant Role of Pterocarpus Marsupiumin in Experimental Hyperglycaemia

Authors: Leelavinothan Pari , Ayyasamy Rathinam

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Diabetes mellitus (DM) is a major and growing public health problem throughout the world. Pterocarpus marsupium (Roxb.) (Family: Fabaceae) is widely used as a traditional medicine to treat various diseases including diabetes. However, the molecular mechanism of Pterocarpus marsupium has not been investigated so far. Two fractions (2.5% and 5%) of extract from the medicinal plant, Pterocarpus marsupium (PME) were conducted in a dose dependent manner in streptozotocin (45 mg/kg b.w.) induced type 2 diabetic rats. Each fraction of PME was administered to diabetic rats intragastrically at a dose of 50, 100 and 200 mg/kg b.w for 45 days. The effective dose 200 mg/kg b.w of 5% fraction was more pronounced in reducing the levels of blood glucose (95.65 mg/dL) and glycosylated hemoglobin (HbA1c) (0.41 mg/g Hb), and increasing the plasma insulin (16.20 µU/mL) level. Moreover, PME (200 mg/kg b.w) significantly ameliorated lipid peroxidation products (thiobarbituric reactive substances, lipid hydroperoxides) enzymatic (superoxide dismutase, catalase and glutathione peroxidase) and non-enzymatic antioxidants (Vitamin C, Vitamin E and reduced glutathione) levels. The altered activities of the key enzymes of lipid metabolism along with the lipid profile in diabetic rats were significantly reverted to near normal levels by the administration of PME 5% 200 mg/kg b.w fraction. PME (200 mg/kg b.w) has the ability to reduce the inflammatory cytokines, such as TNF-α, IL-6 mRNA, as well as protein expression and apoptotic marker, such as caspase-3 enzyme in diabetic hepatic tissue. The above biochemical findings were also supported by histological studies such as improvement in pancreas and liver. Pterocarpus marsupium could effectively reduce the hyperglycemia, oxidative-stress, inflammation and hyperlipedimea in diabetic rats; hence it could be a useful drug in the management of diabetes without any side effects.

Keywords: diabetes mellitus, streptozotocin, Pterocarpus marsupium, lipid peroxidation, Antioxidants, inflammatory cytokines

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572 Allele Mining for Rice Sheath Blight Resistance by Whole-Genome Association Mapping in a Tail-End Population

Authors: Naoki Yamamoto, Hidenobu Ozaki, Taiichiro Ookawa, Youming Liu, Kazunori Okada, Aiping Zheng

Abstract:

Rice sheath blight is one of the destructive fungal diseases in rice. We have thought that rice sheath blight resistance is a polygenic trait. Host-pathogen interactions and secondary metabolites such as lignin and phytoalexins are likely to be involved in defense against R. solani. However, to our knowledge, it is still unknown how sheath blight resistance can be enhanced in rice breeding. To seek for an alternative genetic factor that contribute to sheath blight resistance, we mined relevant allelic variations from rice core collections created in Japan. Based on disease lesion length on detached leaf sheath, we selected 30 varieties of the top tail-end and the bottom tail-end, respectively, from the core collections to perform genome-wide association mapping. Re-sequencing reads for these varieties were used for calling single nucleotide polymorphisms among the 60 varieties to create a SNP panel, which contained 1,137,131 homozygous variant sites after filitering. Association mapping highlighted a locus on the long arm of chromosome 11, which is co-localized with three sheath blight QTLs, qShB11-2-TX, qShB11, and qSBR-11-2. Based on the localization of the trait-associated alleles, we identified an ankyryn repeat-containing protein gene (ANK-M) as an uncharacterized candidate factor for rice sheath blight resistance. Allelic distributions for ANK-M in the whole rice population supported the reliability of trait-allele associations. Gene expression characteristics were checked to evaluiate the functionality of ANK-M. Since an ANK-M homolog (OsPIANK1) in rice seems a basal defense regulator against rice blast and bacterial leaf blight, ANK-M may also play a role in the rice immune system.

Keywords: allele mining, GWAS, QTL, rice sheath blight

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571 The Contribution of Genetic Polymorphisms of Tumor Necrosis Factor Alpha and Vascular Endothelial Growth Factor into the Unfavorable Clinical Course of Ulcerative Colitis

Authors: Y. I. Tretyakova, S. G. Shulkina, T. Y. Kravtsova, A. A. Antipova, N. Y. Kolomeets

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The research aimed to assess the functional significance of tumor necrosis factor-alpha (TNF-α) gene polymorphism at the -308G/A (rs1800629) region and vascular endothelial growth factor A (VEGFA) gene polymorphism at the -634G/C (rs 2010963) region in the development of ulcerative colitis (UC), focusing on patients from the Perm region, Russia. We examined 70 UC patients and 50 healthy donors during the active phase of the disease. Our focus was on TNF-α and VEGF concentration in the blood serum, as well as TNF-α and VEGFA gene polymorphisms at the -308G/А and -634G/C regions, respectively. We found that TNF-α and VEGF levels were significantly higher in patients with severe UC and high endoscopic activity compared to those with milder forms of the disease and low endoscopic activity. These tests could serve as additional non-invasive markers for assessing mucosal damage in the large intestine of UC patients. The frequency of allele variations in the TNF-α gene -308G/A (rs1800629) revealed a significantly higher occurrence of the unfavorable homozygote AA in UC patients compared to donors. Additionally, the major allele G and the allele pair GG were more frequent in patients with mild to moderate disease and 1-2 degree of endoscopic activity than in those with severe UC and 3-4 degree of endoscopic activity (χ2=14.19; p=0.000). We also observed a mutant allele A and the unfavorable homozygote AA associated with severe progressive UC. The occurrence of the mutant allele increased the risk of severe UC by 5 times (OR 5.03; CI 12.07-12.21). We did not find any significant differences in the frequency of the CC homozygote (χ2=1.02; p=0.6; OR=1.32) and the mutant allele C of the VEGFA gene -634G/C (rs 2010963) (χ2=0.01; p=0.913; OR=0.97) between groups of UC patients and healthy individuals. However, we detected that the mutant allele C and the unfavorable homozygote CC of the VEGFA gene were associated with more severe endoscopic changes in the colonic mucosa of UC patients (χ2=25,76; р=0,000; OR=0,15). The presence of the mutant allele increased the risk of severe UC by 6 times (OR 6,78; CI 3,13–14,7). We found a direct correlation between TNF-α and VEGFA gene polymorphisms, increased production of the same factors, disease severity, and endoscopic activity (р=0.000). Therefore, the presence of the mutant allele A and homozygote AA of the TNF-α gene at the -308G/A region and the mutant allele C and homozygote CC of the VEGFA gene at the -634G/C region are associated with risks related to an unfavorable clinical course of UC, frequent recurrences, and rapid progression. These findings should be considered when making prognoses regarding the clinical course of the disease and selecting treatment strategies. The presence of the homozygote AA in the TNF-α gene (rs1800629) is considered a sign of genetic predisposition to UC.

Keywords: gene polymorphism, TNF-α, ulcerative colitis, VEGF

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570 Physical, Microstructural and Functional Quality Improvements of Cassava-Sorghum Composite Snacks

Authors: Adil Basuki Ahza, Michael Liong, Subarna Suryatman

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Healthy chips now dominating the snack market shelves. More than 80% processed snack foods in the market are chips. This research takes the advantages of twin extrusion technology to produce two types of product, i.e. directly expanded and intermediate ready-to-fry or microwavable chips. To improve the functional quality, the cereal-tuber based mix was enriched with antioxidant rich mix of temurui, celery, carrot and isolated soy protein (ISP) powder. Objectives of this research were to find best composite cassava-sorghum ratio, i.e. 60:40, 70:30 and 80:20, to optimize processing conditions of extrusion and study the microstructural, physical and sensorial characteristics of the final products. Optimization was firstly done by applying metering section of extruder barrel temperatures of 120, 130 and 140 °C with screw speeds of 150, 160 and 170 rpm to produce direct expanded product. The intermediate product was extruded in 100 °C and 100 rpm screw speed with feed moisture content of 35, 40 and 45%. The directly expanded products were analyzed for color, hardness, density, microstructure, and organoleptic properties. The results showed that interaction of ratio of cassava-sorghum and cooking methods affected the product's color, hardness, and bulk density (p<0.05). Extrusion processing conditions also significantly affected product's microstructure (p<0.05). The direct expanded snacks of 80:20 cassava-sorghum ratio and fried expanded one 70:30 and 80:20 ratio shown the best organoleptic score (slightly liked) while baking the intermediate product with microwave were resulted sensorial not acceptable quality chips.

Keywords: cassava-sorghum composite, extrusion, microstructure, physical characteristics

Procedia PDF Downloads 278
569 Calcium Release- Activated Calcium Channels as a Target in Treatment of Allergic Asthma

Authors: Martina Šutovská, Marta Jošková, Ivana Kazimierová, Lenka Pappová, Maroš Adamkov, Soňa Fraňová

Abstract:

Bronchial asthma is characterized by increased bronchoconstrictor responses to provoking agonists, airway inflammation and remodeling. All these processes involve Ca2+ influx through Ca2+-release-activated Ca2+ channels (CRAC) that are widely expressed in immune, respiratory epithelium and airway smooth muscle (ASM) cells. Our previous study pointed on possible therapeutic potency of CRAC blockers using experimental guinea pigs asthma model. Presented work analyzed complex anti-asthmatic effect of long-term administered CRAC blocker, including impact on allergic inflammation, airways hyperreactivity, and remodeling and mucociliary clearance. Ovalbumin-induced allergic inflammation of the airways according to Franova et al. was followed by 14 days lasted administration of CRAC blocker (3-fluoropyridine-4-carboxylic acid, FPCA) in the dose 1.5 mg/kg bw. For comparative purposes salbutamol, budesonide and saline were applied to control groups. The anti-inflammatory effect of FPCA was estimated by serum and bronchoalveolar lavage fluid (BALF) changes in IL-4, IL-5, IL-13 and TNF-α analyzed by Bio-Plex® assay as well as immunohistochemical staining focused on assessment of tryptase and c-Fos positivity in pulmonary samples. The in vivo airway hyperreactivity was evaluated by Pennock et al. and by organ tissue bath methods in vitro. The immunohistochemical changes in ASM actin and collagen III layer as well as mucin secretion evaluated anti-remodeling effect of FPCA. The measurement of ciliary beat frequency (CBF) in vitro using LabVIEW™ Software determined impact on mucociliary clearance. Long-term administration of FPCA to sensitized animals resulted in: i. Significant decrease in cytokine levels, tryptase and c-Fos positivity similar to budesonide effect; ii.Meaningful decrease in basal and bronchoconstrictors-induced in vivo and in vitro airway hyperreactivity comparable to salbutamol; iii. Significant inhibition of airway remodeling parameters; iv. Insignificant changes in CBF. All these findings confirmed complex anti-asthmatic effect of CRAC channels blocker and evidenced these structures as the rational target in the treatment of allergic bronchial asthma.

Keywords: allergic asthma, CRAC channels, cytokines, respiratory epithelium

Procedia PDF Downloads 518
568 Plasma Engineered Nanorough Substrates for Stem Cells in vitro Culture

Authors: Melanie Macgregor-Ramiasa, Isabel Hopp, Patricia Murray, Krasimir Vasilev

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Stem cells based therapies are one of the greatest promises of new-age medicine due to their potential to help curing most dreaded conditions such as cancer, diabetes and even auto-immune disease. However, establishing suitable in vitro culture materials allowing to control the fate of stem cells remain a challenge. Amongst the factor influencing stem cell behavior, substrate chemistry and nanotopogaphy are particularly critical. In this work, we used plasma assisted surface modification methods to produce model substrates with tailored nanotopography and controlled chemistry. Three different sizes of gold nanoparticles were bound to amine rich plasma polymer layers to produce homogeneous and gradient surface nanotopographies. The outer chemistry of the substrate was kept constant for all substrates by depositing a thin layer of our patented biocompatible polyoxazoline plasma polymer on top of the nanofeatures. For the first time, protein adsorption and stem cell behaviour (mouse kidney stem cells and mesenchymal stem cells) were evaluated on nanorough plasma deposited polyoxazoline thin films. Compared to other nitrogen rich coatings, polyoxazoline plasma polymer supports the covalent binding of proteins. Moderate surface nanoroughness, in both size and density, triggers cell proliferation. In association with polyoxazoline coating, cell proliferation is further enhanced on nanorough substrates. Results are discussed in term of substrates wetting properties. These findings provide valuable insights on the mechanisms governing the interactions between stem cells and their growth support.

Keywords: nanotopography, stem cells, differentiation, plasma polymer, oxazoline, gold nanoparticles

Procedia PDF Downloads 276
567 Economics of Fish-Plantain Integrated Farm Enterprise in Southern Nigeria

Authors: S. O. Obasa, J. A. Soaga, O. I. Afolabi, N. A. Bamidele, O. E. Babalola

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Attempt to improve the income of the rural population is a welcome development in Nigeria. Integrated fish-crop farming has been suggested as a means of raising farm income, reducing wastage and mitigating the risk component in production through the complementarity gain. A feeding trial was carried out to investigate the replacement of maize with fermented unripe plantain (Musa paradisiaca) peel meal in the diet of Nile tilapia, Oreochromis niloticus. The economics of the integrated enterprise was assessed using budgetary analysis techniques. The analysis incorporated the material and labour costs as well as the returns from sale of matured fish and plantain. A total of 60 fingerlings of Nile tilapia (1.70±0.1 g) were stocked at 10 per plastic tank. Two iso-nitrogenous diets containing 35% crude protein in which maize meal was replaced by fermented unripe plantain peel meal at 0% (FUP0/Control diet), and 100% (FUP100) were formulated and prepared. The fingerlings were fed at 5% body weight per day for 56 days. Lowest feed conversion ratio of 1.39 in fish fed diet FUP100 was not significantly different (P > 0.05) from the highest 1.42 of fish fed the Control diet. The highest percentage profit of 88.85% in fish fed diet FUP100 was significantly higher than 66.68% in fish fed diet FUP0, while the profit index of 1.89 in fish fed diet FUP100 was significantly different from 1.67 in fish fed diet FUP0. Therefore, fermented unripe plantain peel meal can completely replace maize in the diet of O. niloticus fingerlings. Profitability assessment shows that the net income from the integration was ₦ 463,000 per hectare and the integration resulted to an increase of ₦ 87,750.00 representing a 12.2% increase than in separate production.

Keywords: fish-crop, income, Nile tilapia, waste management

Procedia PDF Downloads 495
566 Interaction of GCN5L1 with WHAMM and KIF5B Regulates Autolysosome Tubulation

Authors: Allen Seylani

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Lysosome-dependent autophagy is a nutrient-deprivation-induced evolutionarily conserved intracellular recycling program that sequestrates intracellular cargo into autophagosomes (AP), which then fuse with lysosomes to form autolysosomes (ALs) for cargo digestion. To restore free lysosomes, autophagic lysosome reformation (ALR) is initiated by extrusion of tubular structures from autolysosomes at the final stage of autophagy, in a process called lysosomal tubulation (LT). This project aimed to investigate the molecular mechanism of GCN5L1 in LT and the following lysosomal signaling. GCN5L1 belongs to the BORC multiprotein complexes and is involved in controlling lysosomal trafficking; however, the effect of GCN5L1 on lysosome tubulation remains largely unknown. Genetic ablation of GCN5L1 in the mouse primary hepatocytes showed dramatically increased autolysosomes (ALs), decreased lysosome regeneration and absence of lysosomal tubulation. This phenotype suggests the possibility of disruption in lysosome tubulation, which results in the disturbance of the overall lysosome homeostasis. The formation of tubulars from ALs requires kinesin motor protein KIF5B. Immunoprecipitation was employed and confirmed the interaction of GCN5L1 with the ARL8B-KIF5B complex, which recruited KIF5B to ALs. At the same time, GCN5L1 interacted with WHAMM, which promotes the actin nucleation factor, which brings actin cytoskeleton to ALs and initiates LT. Furthermore, impaired LT in GCN5L1 deficient hepatocytes was restored by overexpression of GCN5L1, and this rescue effect was attenuated by knockdown of KIF5B. Additionally, lysosomal mTORC1 activity was upregulated in GCN5L1-/- hepatocytes, while inhibition of mTORC1 abrogated the GCN5L1 mediated rescue of LT in knockout hepatocytes. Altogether these findings revealed a novel mechanism of ALR, in which a simultaneous interaction of GCN5L1 with KIF5B and WHAMM is required for LT and downstream mTORC1 signaling.

Keywords: autophagy, autolysosome, GCN5L1, lysosome

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565 Influence of Dietary Herbal Blend on Crop Filling, Growth Performance and Nutrient Digestibility in Broiler Chickens

Authors: S. Ahmad, M. Rizwan, B. Ayub, S. Mehmood, P. Akhtar

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This experiment was conducted to investigate the effect of supplementation of pure herbal blend on growth performance of boilers. One hundred and twenty birds were randomly distributed into 4 experimental units of 3 replicates (10 birds/replicate) as: negative control (basal diet), positive control (Lincomycin at the rate of 5g/bag), pure herbal blend at the rate of 150g/bag and pure herbal blend at the rate of 300g/bag. The data regarding weekly feed intake, body weight gain and feed conversion ratio were recorded, and fecal samples were collected at the end of starter and finisher phase for nutrient digestibility trial. The results of feed intake showed significant (P < 0.05) results in 1st (305g), 2nd (696.88g), 3rd (1046.9g) and 4th (1173.2g) week and feed conversion ratio indicated significant (P < 0.05) variations in 1st (2.54) and 4th (2.28) week of age. Also, both starter and finisher phase indicated significant (P < 0.05) differences among all treatment groups in feed intake (2023.4g) and (2302.6g) respectively. The statistical analysis indicated significant (P < 0.05) results in crop filling percentage (86.6%) after 2 hours of first feed supplementation. In case of nutrient digestibility trial, results showed significant (P < 0.05) values of crude protein and crude fat in starter phase as 69.65% and 56.62% respectively, and 69.57% and 48.55% respectively, in finisher phase. Based on overall results, it was concluded that the dietary inclusion of pure herbal blend containing neem tree leaves powder, garlic powder, ginger powder and turmeric powder increase the production performance of broilers.

Keywords: neem tree leave, garlic, ginger, herbal blend, broiler

Procedia PDF Downloads 198
564 Cerebrum Maturity Damage Induced by Fluoride in Suckling Mice

Authors: Hanen Bouaziz, Françoise Croute, Najiba Zeghal

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In order to investigate the toxic effects of fluoride on cerebrum maturity of suckling mice, we treated adult female mice of Swiss Albinos strain by 500 ppm NaF in their drinking water from the 15th day of pregnancy until the day 14 after delivery. All mice were sacrificed on day 14 after parturition. During treatment, levels of thiobarbituric acid reactive substances, the marker of lipid peroxidation extend, increased, while the activities of the antioxidant enzymes such as glutathione peroxidase, superoxide dismutase and catalase and the level of glutathione decreased significantly in cerebellum compared with those of the control group. These results suggested that fluoride enhanced oxidative stress, thereby disturbing the antioxidant defense of nursing pups. In addition, acetylcholinesterase activity in cerebellum was inhibited after treatment with fluoride. In cerebellum of mice, migration of neurons from the external granular layer to the internal granular layer occurred postnatally. Key guidance signals to these migrating neurons were provided by laminin, an extracellular matrix protein fixed to the surface of astrocytes. In the present study, we examined the expression and distribution of laminin in cerebellum of 14-day-old mice. Immunoreactive laminin was disappeared by postnatal day 14 in cerebellum parenchyma of control pups and was restricted to vasculature despite the continued presence of granular cells in the external granular layer. In contrast, in cerebellum of NaF treated pups, laminin was deposited in organised punctuate clusters in the molecular layer. These data indicated that the disruption of laminin distribution might play a major role in the profound derangement of neuronal migration observed in cerebellum of NaF treated pups.

Keywords: acetylcholinesterase activity, cerebellum, laminin, oxidative stress, suckling mice

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563 Biochemical and Pomological Variability among 14 Moroccan and Foreign Cultivars of Prunus dulcis

Authors: H. Hanine, H. H'ssaini, M. Ibno Alaoui, A. Nablousi, H. Zahir, S. Ennahli, H. Latrache, H. Zine Abidine

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Biochemical and pomological variability among 14 cultivars of Prunus dulcis planted in a germoplasm collection site in Morocco were evaluated. Almond samples from six local and eight foreign cultivars (France, Italy, Spain, and USA) were characterized. Biochemical and pomological data revealed significant genetic variability among the 14 cultivars; local cultivars exhibited higher total polyphenol content. Oil content ranged from 35 to 57% among cultivars; both Texas and Toundout genotypes recorded the highest oil content. Total protein concentration from select cultivars ranged from 50 mg/g in Ferraduel to 105 mg/g in Rizlane1 cultivars. Antioxidant activity of almond samples was examined by a DPPH (1,1-diphenyl-2-picrylhydrazyl) radical-scavenging assay; the antioxidant activity varied significantly within the cultivars, with IC50 (the half-maximal inhibitory concentration) values ranging from 2.25 to 20 mg/ml. Autochthonous cultivars originated from the Oujda region exhibited higher tegument total polyphenol and amino acid content compared to others. The genotype Rizlane2 recorded the highest flavonoid content. Pomological traits revealed a large variability within the almond germplasms. The hierarchical clustering analysis of all the data regarding pomological traits distinguished two groups with some particular genotypes as distinct cultivars, and groups of cultivars as polyclone varieties. These results strongly exhibit a potential use of Moroccan-originated almonds as potential clones for future selection due to their nutritional values and pomological traits compared to well-established cultivars.

Keywords: antioxidant activity, DDPH, Moroccan almonds, Prunus dulcis

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562 TNF-Alpha and MDA Levels in Hearts of Cholesterol-Fed Rats Supplemented with Extra Virgin Olive Oil or Sunflower Oil, in Either Commercial or Modified Forms

Authors: Ageliki I. Katsarou, Andriana C. Kaliora, Antonia Chiou, Apostolos Papalois, Nick Kalogeropoulos, Nikolaos K. Andrikopoulos

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Oxidative stress is a major mechanism underlying CVDs while inflammation, an intertwined process with oxidative stress, is also linked to CVDs. Extra virgin olive oil (EVOO) is widely known to play a pivotal role in CVD prevention and CVD reduction. However, in most studies, olive oil constituents are evaluated individually and not as part of the native food, hence potential synergistic effects as drivers of EVOO beneficial properties may be underestimated. In this study, EVOO lipidic and polar phenolics fractions were evaluated for their effect on inflammatory (TNF-alpha) and oxidation (malondialdehyde/MDA) markers, in cholesterol-fed rats. Thereat, oils with discernible lipidic profile and polar phenolic content were used. Wistar rats were fed on either a high-cholesterol diet (HCD) or a HCD supplemented with oils, either commercially available, i.e. EVOO, sunflower oil (SO), or modified as to their polar phenol content, i.e. phenolics deprived-EVOO (EVOOd), SO enriched with the EVOO phenolics (SOe). After 9 weeks of dietary intervention, heart and blood samples were collected. HCD induced dylipidemia shown by increase in serum total cholesterol, low-density lipoprotein cholesterol (LDL-c) and triacylglycerols. Heart tissue has been affected by dyslipidemia; oxidation was indicated by increase in MDA in cholesterol-fed rats and inflammation by increase in TNF-alpha. In both cases, this augmentation was attenuated in EVOO and SOe diets. With respect to oxidation, SO enrichment with the EVOO phenolics brought its lipid peroxidation levels as low as in EVOO-fed rats. This suggests that phenolic compounds may act as antioxidant agents in rat heart. A possible mechanism underlying this activity may be the protective effect of phenolics in mitochondrial membrane against oxidative damage. This was further supported by EVOO/EVOOd comparison with the former presenting lower heart MDA content. As for heart inflammation, phenolics naturally present in EVOO as well as phenolics chemically added in SO, exhibited quenching abilities in heart TNF-alpha levels of cholesterol-fed rats. TNF-alpha may have played a causative role in oxidative stress induction while the opposite may have also happened, hence setting up a vicious cycle. Overall, diet supplementation with EVOO or SOe attenuated hypercholesterolemia-induced increase in MDA and TNF-alpha in Wistar rat hearts. This is attributed to phenolic compounds either naturally existing in olive oil or as fortificants in seed oil.

Keywords: extra virgin olive oil, hypercholesterolemic rats, MDA, polar phenolics, TNF-alpha

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561 Impact of Length of Straw by the Use of a Straw Mill on the Selective Feeding of Young Cattle and Their Effects for the Cattle

Authors: Heiko Scholz

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When feeding high qualitysilagetoheifersfromthe age of two, there is a riskofenergyoversupply. Depending on the feeding valueorscarceavailability ofsilageorcorn silage diets withhighproportionsof straw is often incorporated. Foran energetically standardized young cattle supply of strawproportion can be more than 20% of dry matter. It was investigated whether the grinding of straw with the strawmillselective feeding significantly limits. The investigation has been carried out with young cattle in the second year. 78 animals were kept and fed under similar conditions in two groups. The experimental group (EG) consisted of cattle 12 to 15 months, and in the control group (CG), the cattle were 15 to 20 months old. The experimental feeding took place in five days of feed distribution, and residual feed were weighed. The ration of EG contained ground with the straw mill straw, and CG was further fed rotor-cut pressed straw. To determine the selective seizure samples of feed distributionandtheremainingfood with the particle separator boxandthecrude protein-and energy-content have been determined. The grinding of the straw increased the daily feed intake.IntheEGan increase infeed intakewas observedby grinding of the straw. Feed intakedirectlyon the day for changing the dietoflongonground straw increased by more than 2.0 kgofDMper animal. In the following days, the feed intakewasincreasedby 0.9kg DMper animal and day on average (7.4 vs. 8.3 kg DM per day). The results of the screen distribution of residual feed point to a differentiated feeding behavior between the groups. In the EG, the particle length of the residual feed to a large extent with the template matches. The acid-base-balance (NSBA)valuesofEGarewithin normal limits. Ifstrawsharesof25% and more are federations to young cattle (heifers), the theparticlelengthof straw has significant impact ontheselectivefeeding behavior. Aparticlelength of 1.5cmcompared to7.5 cmlongpreventedstrawcertainly discarding of the straw on the feeding barn. The feed intake increases whenshortstrawis mixed into theTMR.

Keywords: straw mill, heifer, feed selection, dry matter intake

Procedia PDF Downloads 193
560 Design and Optimisation of 2-Oxoglutarate Dioxygenase Expression in Escherichia coli Strains for Production of Bioethylene from Crude Glycerol

Authors: Idan Chiyanzu, Maruping Mangena

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Crude glycerol, a major by-product from the transesterification of triacylglycerides with alcohol to biodiesel, is known to have a broad range of applications. For example, its bioconversion can afford a wide range of chemicals including alcohols, organic acids, hydrogen, solvents and intermediate compounds. In bacteria, the 2-oxoglutarate dioxygenase (2-OGD) enzymes are widely found among the Pseudomonas syringae species and have been recognized with an emerging importance in ethylene formation. However, the use of optimized enzyme function in recombinant systems for crude glycerol conversion to ethylene is still not been reported. The present study investigated the production of ethylene from crude glycerol using engineered E. coli MG1655 and JM109 strains. Ethylene production with an optimized expression system for 2-OGD in E. coli using a codon optimized construct of the ethylene-forming gene was studied. The codon-optimization resulted in a 20-fold increase of protein production and thus an enhanced production of the ethylene gas. For a reliable bioreactor performance, the effect of temperature, fermentation time, pH, substrate concentration, the concentration of methanol, concentration of potassium hydroxide and media supplements on ethylene yield was investigated. The results demonstrate that the recombinant enzyme can be used for future studies to exploit the conversion of low-priced crude glycerol into advanced value products like light olefins, and tools including recombineering techniques for DNA, molecular biology, and bioengineering can be used to allowing unlimited the production of ethylene directly from the fermentation of crude glycerol. It can be concluded that recombinant E.coli production systems represent significantly secure, renewable and environmentally safe alternative to thermochemical approach to ethylene production.

Keywords: crude glycerol, bioethylene, recombinant E. coli, optimization

Procedia PDF Downloads 277
559 Surface-Enhanced Raman Spectroscopy-Based Detection of SARS-CoV-2 Through In Situ One-pot Electrochemical Synthesis of 3D Au-Lysate Nanocomposite Structures on Plasmonic Au Electrodes

Authors: Ansah Iris Baffour, Dong-Ho Kim, Sung-Gyu Park

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The ongoing COVID-19 pandemic, caused by the SARS-CoV-2 virus and is gradually shifting to an endemic phase which implies the outbreak is far from over and will be difficult to eradicate. Global cooperation has led to unified precautions that aim to suppress epidemiological spread (e.g., through travel restrictions) and reach herd immunity (through vaccinations); however, the primary strategy to restrain the spread of the virus in mass populations relies on screening protocols that enable rapid on-site diagnosis of infections. Herein, we employed surface enhanced Raman spectroscopy (SERS) for the rapid detection of SARS-CoV-2 lysate on an Au-modified Au nanodimple(AuND)electrode. Through in situone-pot Au electrodeposition on the AuND electrode, Au-lysate nanocomposites were synthesized, generating3D internal hotspots for large SERS signal enhancements within 30 s of the deposition. The capture of lysate into newly generated plasmonic nanogaps within the nanocomposite structures enhanced metal-spike protein contact in 3D spaces and served as hotspots for sensitive detection. The limit of detection of SARS-CoV-2 lysate was 5 x 10-2 PFU/mL. Interestingly, ultrasensitive detection of the lysates of influenza A/H1N1 and respiratory syncytial virus (RSV) was possible, but the method showed ultimate selectivity for SARS-CoV-2 in lysate solution mixtures. We investigated the practical application of the approach for rapid on-site diagnosis by detecting SARS-CoV-2 lysate spiked in normal human saliva at ultralow concentrations. The results presented demonstrate the reliability and sensitivity of the assay for rapid diagnosis of COVID-19.

Keywords: label-free detection, nanocomposites, SARS-CoV-2, surface-enhanced raman spectroscopy

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558 Effect of Diet Inulin Prebiotic on Growth, Reproductive Performance, Carcass Composition and Resistance to Environmental Stresses in Zebra Danio (Danio rerio)

Authors: Ehsan Ahmadifar

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In this research, the effects of different levels (control group (T0), (T1)1, (T2)2 and (T3)3 gr Inulin per Kg diet) of prebiotic Inulin as nutritional supplement on Danio rerio were investigated for 4 month. Since the beginning of feeding larvae until adult (average weight: 67.1 g, length: 4.5 cm) were fed with experimental diets. The survival rate of fish had no significant effect on rate survival (P > 0.05). The highest food conversion ratio (FCR) was in control group and the lowest was observed in T3. Treatment of T3 significantly caused the best feed conversion ratio in Zebra fish (P < 0.05). By increasing the inulin diet during the experiment, specific growth rate increased. The highest and the lowest body weight gain and condition factor were observed in T3 and control, respectively (P < 0.05). Adding 3 gr inulin in Zebra fish diet can improve the performance of the growth indices and final biomass, also this prebiotic can be considered as a suitable supplement for Cyprinidae diet. In the first sampling stage for feeding fish, fat and muscle protein was significantly higher than the second sampling stage (P < 0.05). Given that the second stage fish were full sexual maturity, the amount of fat in muscle decreased (P < 0.05). Moisture and ash levels were significantly (P < 0.05) higher in the second stage sampling than the first stage. Overall, different stage of living affected on muscle chemical composition muscle. Reproductive performance in treatment T2 and T3 were significantly higher than other treatments (P < 0.05). According to the results, the prebiotic inulin does not have a significant impact on the sex ratio in zebrafish (P > 0.05). Based on histology of the gonads, the use of dietary inulin accelerates the process of gonad development in zebrafish.

Keywords: inulin, zebrafish, reproduction, histology

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557 Inadequate Intake of Energy and Nutrients: A Comparative Cross-Sectional Study Between Sport and Non-sport Science University Students of Southern Ethiopia

Authors: Beruk Berhanu Desalegn, Kebede Awgechew, Addisalem Mesfin

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Introduction: This study aimed to investigate and compare the energy and selected nutrient intakes of sport science and non-sport science University students of Southern Ethiopia. Method: Multiple-day dietary data were collected from 166 university students (76 sport science and 90 non-sport sciences). Average daily energy and nutrient intake, and inadequate intakes were calculated using NutriSurvey (NS). Results: There were significant differences (p < 0.05) in the median intakes of energy, total carbohydrate, and vitamin B1 between female students from the sport science and non-sport science groups, but only the median intake of iron was significantly different (p < 0.05) between the male sport and non-sport science students’ group. The prevalence of inadequate intake of vitamin B1 were significantly (p<0.05) higher in the male and female from the non-sport science groups compared to the male and female students’ groups in the sport science, respectively. Whereas, the prevalence of inadequate iron intake by the male sport science students’ group was significantly (p<0.05) higher compared to their counterparts. Similarly, the prevalence of inadequate energy among the females from the sport science group was significantly (p<0.05) higher compared to the female students from the non-sport science department group. The prevalence of inadequate intakes of dietary energy, and the majority of the nutrients (protein, fat, vitamin A, B1, B2, and magnesium) were high (>50%) in selected University students. Conclusion: The energy and majority of nutrient intakes by the students in the selected universities of southern Ethiopia were sub-optimal. Therefore, activities that will improve the dietary intake of University students should include weekly meal plan revision considering their average recommended nutrient intake (RNI).

Keywords: dietary intake, sport science, University students, Ethiopia

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556 Acupoint Injection of High Concentration of Glucose Attenuates Mice Chronic Pain and Depression Comorbidity

Authors: Chanya Inprasit, Yi-Wen Lin

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Inflammation causes changes of peripheral and central nervous system properties, affecting both neuronal and non-neuronal cells, resulting in inflammatory pain. Acupoint injection (AI) was developed in the 1950s and has been widely used for relieving pain. It is an acupoint-stimulating technique that utilizes anatomically based meridians derived from Chinese medicine theory. AI has been accepted as an effective treatment and is thought to display superior results when compared to traditional acupuncture methods. However, the mechanism of AI needs to be ratified by more scientific evidence in order to support the theory and its therapeutic development. In this study, we explored the effect of AI on the comorbidity of chronic pain and depression. Mice hindpaw was injected by complete Freund’s adjuvant (CFA) to induce the condition of chronic pain. Measurements of mechanical and thermal hyperalgesia and depression-like behavior were analyzed. The results indicated a positive tendency to AI treatment. The comorbidity of chronic pain and depression was investigated with relation to transient receptor potential V1 (TRPV1) mechanism through the use of TRPV1 gene deletion. The expression of nociceptors such as voltage-gated sodium channels (Navs) or TRPV1, was significantly down-regulated by AI. The expression of inflammation-activated molecules: astrocytic marker glial fibrillary acidic protein (GFAP), the microglial marker Iba-1, S100B, and related kinases, were reversed by AI in both the peripheral and central nervous system. Taken together, these data provided a detailed molecular mechanism of AI-induced analgesia and anti-inflammatory properties. This finding may be utilized for clinical practice to treat chronic pain and depression comorbidity.

Keywords: inflammatory pain, acupoint injection, TRPV1, GFAP, S100B

Procedia PDF Downloads 146