Search results for: immunohistochemical markers
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 817

Search results for: immunohistochemical markers

337 Zebrafish Larvae Model: A High Throughput Screening Tool to Study Autism

Authors: Shubham Dwivedi, Raghavender Medishetti, Rita Rani, Aarti Sevilimedu, Pushkar Kulkarni, Yogeeswari Perumal

Abstract:

Autism Spectrum Disorder (ASD) is a complex neurodevelopmental disorder of early onset, characterized by impaired sociability, cognitive function and stereotypies. There is a significant urge to develop and establish new animal models with ASD-like characteristics for better understanding of underlying mechanisms. The aim of the present study was to develop a cost and time effective zebrafish model with quantifiable parameters to facilitate mechanistic studies as well as high-throughput screening of new molecules for autism. Zebrafish embryos were treated with valproic acid and a battery of behavioral tests (anxiety, inattentive behavior, irritability and social impairment) was performed on larvae at 7th day post fertilization, followed by study of molecular markers of autism. This model shows a significant behavioural impairment in valproic acid treated larvae in comparison to control which was again supported by alteration in few marker genes and proteins of autism. The model also shows a rescue of behavioural despair with positive control drugs. The model shows robust parameters to study behavior, molecular mechanism and drug screening approach in a single frame. Thus we postulate that our 7 days zebrafish larval model for autism can help in high throughput screening of new molecules on autism.

Keywords: autism, zebrafish, valproic acid, neurodevelopment, behavioral assay

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336 Polymorphisms of STAT5A and DGAT1 Genes and Their Associations with Milk Trait in Egyptian Goats

Authors: Othman Elmahdy Othman

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The objectives of this study were to identify polymorphisms in the STAT5A using Restriction Fragment Length Polymorphism and DGAT1 using Single-Strand Conformation Polymorphism genes among three Egyptian goat breeds (Barki, Zaraibi, and Damascus) as well as investigate the effect of their genotypes on milk composition traits of Zaraibi goats. One hundred and fifty blood samples were collected for DNA extraction, 60 from Zaraibi, 40 from Damascus and 50 from Barki breeds. Fat, protein and lactose percentages were determined in Zaraibi goat milk using an automatic milk analyzer. Two genotypes, CC and CT (for STAT5A) and C-C- and C-C+ (for DGAT1), were identified in the three Egyptian goat breeds with different frequencies. The associations between these genotypes and milk fat, protein and lactose were determined in Zaraibi breed. The results showed that the STAT5A genotypes had significant effects on milk yield, protein, fat and lactose with the superiority of CT genotype over CC. Regarding DGAT1 polymorphism, the result showed the only association between it with milk fat where the animals with C-C+ genotype had greater milk fat than animals possess C-C- genotype. The association of combined genotypes with milk trait declared that the does with heterozygous genotypes for both genes are preferred than does with homozygous genotypes where the animals with CTC-C+ have more milk yield, fat and protein than those with CCC-C- genotype. In conclusion, the result showed that C/T and C-/C+ SNPs of STAT5A and DGAT1 genes respectively may be useful markers for assisted selection programs to improve goat milk composition

Keywords: DGAT1, genetic polymorphism, milk trait, STAT5A

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335 Pharmacological Mechanisms of an Indolic Compound in Chemoprevention of Colonic Acf Formation in Azoxymethane-Induced Colon Cancer Rat Model and Cell Lines

Authors: Nima Samie, Sekaran Muniandy, Zahurin Mohamed, M. S. Kanthimathi

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Although number of indole containing compounds have been reported to have anticancer properties in vitro but only a few of them show potential as anticancer compounds in vivo. The current study was to evaluate the mechanism of cytotoxicity of selected indolic compound in vivo and in vitro. In this context, we determined the potency of the compound in the induction of apoptosis, cell cycle arrest, and cytoskeleton rearrangement. HT-29, WiDr, CCD-18Co, human monocyte/macrophage CRL-9855, and B lymphocyte CCL-156 cell lines were used to determine the IC50 of the compound using the MTT assay. Analysis of apoptosis was carried out using immunofluorescence, acridine orange/ propidium iodide double staining, Annexin-V-FITC assay, evaluation of the translocation of NF-kB, oxygen radical antioxidant capacity, quenching of reactive oxygen species content, measurement of LDH release, caspase-3/-7, -8 and -9 assays and western blotting. The cell cycle arrest was examined using flowcytometry and gene expression was assessed using qPCR array. Results displayed a potent suppressive effect on HT-29 and WiDr after 24 h of treatment with IC50 value of 2.52±0.34 µg/ml and 2.13±0.65 µg/ml respectively. This cytotoxic effect on normal, monocyte/macrophage and B-cells was insignificant. Dipping in the mitochondrial membrane potential and increased release of cytochrome c from the mitochondria indicated induction of the intrinsic apoptosis pathway by the compound. Activation of this pathway was further evidenced by significant activation of caspase-9 and 3/7. The compound was also shown to activate the extrinsic pathways of apoptosis via activation of caspase-8 which is linked to the suppression of NF-kB translocation to the nucleus. Cell cycle arrest in the G1 phase and up-regulation of glutathione reductase, based on excessive ROS production were also observed. These findings were further investigated for inhibitory efficiency of the compound on colonic aberrant crypt foci in male rats. Rats were divided in to 5 groups: vehicle, cancer control, positive control groups and the groups treated with 25 and 50 mg/kg of compounds for 10 weeks. Administration of compound suppressed total colonic ACF formation up to 73.4%. The results also showed that treatment with the compound significantly reduced the level of malondialdehyde while increasing superoxide dismutase and catalase activities. Furthermore, the down-regulation of PCNA and Bcl2 and the up-regulation of Bax was confirmed by immunohistochemical staining. The outcome of this study suggest sthat the indolic compound is a potent anti-cancer agent against colon cancer and can be further evaluated by animal trial.

Keywords: indolic compound, chemoprevention, crypt, azoxymethane, colon cancer

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334 The Effects of Periostin in a Rat Model of Isoproterenol-Mediated Cardiotoxicity

Authors: Mahmut Sozmen, Alparslan Kadir Devrim, Yonca Betil Kabak, Tuba Devrim

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Acute myocardial infarction is the leading cause of deaths in the worldwide. Mature cardiomyocytes do not have the ability to regenerate instead fibrous tissue proliferate and granulation tissue to fill out. Periostin is an extracellular matrix protein from fasciclin family and it plays an important role in the cell adhesion, migration, and growth of the organism. Periostin prevents apoptosis while stimulating cardiomyocytes. The main objective of this project is to investigate the effects of the recombinant murine periostin peptide administration for the cardiomyocyte regeneration in a rat model of acute myocardial infarction. The experiment was performed on 84 male rats (6 months old) in 4 group each contains 21 rats. Saline applied subcutaneously (1 ml/kg) two times with 24 hours intervals to the rats in control group (Group 1). Recombinant periostin peptide (1 μg/kg) dissolved in saline applied intraperitoneally in group 2 on 1, 3, 7, 14 and 21. days on same dates in group 4. Isoproterenol dissolved in saline applied intraperitoneally (85mg/kg/day) two times with 24 hours intervals to the groups 3 and 4. Rats in group 4 further received recombinant periostin peptide (1 μg/kg) dissolved in saline intraperitoneally starting one day after the final isoproterenol administration on days 1, 3, 7, 14 and 21. Following the final application of periostin rats continued to feed routinely with pelleted chow and water ad libitum for further seven days. At the end of 7th day rats sacrificed, blood and heart tissue samples collected for the immunohistochemical and biochemical analysis. Angiogenesis in response to tissue damage, is a highly dynamic process regulated by signals from the surrounding extracellular matrix and blood serum. In this project, VEGF, ANGPT, bFGF, TGFβ are the key factors that contribute to cardiomyocyte regeneration were investigated. Additionally, the relationship between mitosis and apoptosis (Bcl-2, Bax, PCNA, Ki-67, Phopho-Histone H3), cell cycle activators and inhibitors (Cyclin D1, D2, A2, Cdc2), the origin of regenerating cells (cKit and CD45) were examined. Present results revealed that periostin stimulated cardiomyocye cell-cycle re-entry in both normal and MCA damaged cardiomyocytes and increased angiogenesis. Thus, periostin contributes to cardiomyocyte regeneration during the healing period following myocardial infarction which provides a better understanding of its role of this mechanism, improving recovery rates and it is expected to contribute the lack of literature on this subject. Acknowledgement: This project was financially supported by Turkish Scientific Research Council- Agriculture, Forestry and Veterinary Research Support Group (TUBİTAK-TOVAG; Project No: 114O734), Ankara, TURKEY.

Keywords: cardiotoxicity, immunohistochemistry, isoproterenol, periostin

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333 Correlation between Overweightness and the Extent of Coronary Atherosclerosis among the South Caspian Population

Authors: Maryam Nabati, Mahmood Moosazadeh, Ehsan Soroosh, Hanieh Shiraj, Mahnaneh Gholami, Ali Ghaemian

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Background: Reported effects of obesity on the extent of angiographic coronary artery disease(CAD) have beeninconsistent. The present study aimed to investigate the relationships between the indices of obesity and otheranthropometric markers with the extent of CAD. Methods: This study was conducted on 1008 consecutive patients who underwent coronary angiography. Bodymass index (BMI), waist circumference (WC), waist-to-hip ratio (WHR), and waist-to-height ratio (WHtR) wereseparately calculated for each patient. Extent, severity, and complexity of CAD were determined by the Gensini andSYNTAX scores. Results: According to the results, there was a significant inverse correlation between the SYNTAX score with BMI(r = − 0.110; P < 0.001), WC (r = − 0.074; P = 0.018), and WHtR (r = − 0.089; P = 0.005). Furthermore, a significant inversecorrelation was observed between the Gensini score with BMI (r = − 0.090; P = 0.004) and WHtR (r = − 0.065; P =0.041). However, the results of multivariate linear regression analysis did not show any association between theSYNTAX and Gensini scores with the indices of obesity and overweight. On the other hand, the patients with anunhealthy WC had a higher prevalence of diabetes mellitus (DM) (P = 0.004) and hypertension (HTN) (P < 0.001) compared to the patients with healthy values. Coexistence of HTN and DM was more prevalent in subjects with anunhealthy WC and WHR compared to that in those with healthy values (P = 0.002 and P = 0.032, respectively). Conclusion: It seems that the anthropometric indices of obesity are not the predictors of the angiographic severityof CAD. However, they are associated with an increased risk of cardiovascular risk factors and higher risk profile.

Keywords: body mass index, BMI, coronary artery disease, waist circumference

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332 Prospective Validation of the FibroTest Score in Assessing Liver Fibrosis in Hepatitis C Infection with Genotype 4

Authors: G. Shiha, S. Seif, W. Samir, K. Zalata

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Prospective Validation of the FibroTest Score in assessing Liver Fibrosis in Hepatitis C Infection with Genotype 4 FibroTest (FT) is non-invasive score of liver fibrosis that combines the quantitative results of 5 serum biochemical markers (alpha-2-macroglobulin, haptoglobin, apolipoprotein A1, gamma glutamyl transpeptidase (GGT) and bilirubin) and adjusted with the patient's age and sex in a patented algorithm to generate a measure of fibrosis. FT has been validated in patients with chronic hepatitis C (CHC) (Halfon et al., Gastroenterol. Clin Biol.( 2008), 32 6suppl 1, 22-39). The validation of fibro test ( FT) in genotype IV is not well studied. Our aim was to evaluate the performance of FibroTest in an independent prospective cohort of hepatitis C patients with genotype 4. Subject was 122 patients with CHC. All liver biopsies were scored using METAVIR system. Our fibrosis score(FT) were measured, and the performance of the cut-off score were done using ROC curve. Among patients with advanced fibrosis, the FT was identically matched with the liver biopsy in 18.6%, overestimated the stage of fibrosis in 44.2% and underestimated the stage of fibrosis in 37.7% of cases. Also in patients with no/mild fibrosis, identical matching was detected in 39.2% of cases with overestimation in 48.1% and underestimation in 12.7%. So, the overall results of the test were identical matching, overestimation and underestimation in 32%, 46.7% and 21.3% respectively. Using ROC curve it was found that (FT) at the cut-off point of 0.555 could discriminate early from advanced stages of fibrosis with an area under ROC curve (AUC) of 0.72, sensitivity of 65%, specificity of 69%, PPV of 68%, NPV of 66% and accuracy of 67%. As FibroTest Score overestimates the stage of advanced fibrosis, it should not be considered as a reliable surrogate for liver biopsy in hepatitis C infection with genotype 4.

Keywords: fibrotest, chronic Hepatitis C, genotype 4, liver biopsy

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331 Trehalose-Based Nanocarriers for Alleviation of Inflammation in Colitis

Authors: Wessam H. Abd-Elsalam, Mona M. Saber, Samar M. Abouelatta

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Non-steroidal anti-inflammatory drugs (NSAIDs) are considered a double edged sword in inflammatory bowel diseases (IBDs). Some studies reported their advantageous effect in decreasing inflammation, and other studies reported that their use is associated with colitis aggravation. This study aimed to use specifically formulated trehalose-based nano-carriers that targets the colon in an attempt to alleviate inflammation caused by NSAIDs. L-α-phosphatidylcholine (PL), trehalose, and transcutol were used to prepare the trehalosomes (THs), which were also loaded with Tenoxicam(TXM) as a model NSAID. To optimize the formulation variables, a full 23 factorial design, using Design-Expert® software, was performed. The optimized formulation composed of trehalose: PL at a weight ratio of 1:1, 377.72 mg transcutol, and sonicated for 4 min, possessed a spherical shape with a size of 268.61 nm and EE% of 97.83% and released 70.22% of its drug content over 24 h. The superior protective action of TXM loaded THs compared to TXM suspension and drug-free THs was shown by the inhibition of the inflammatory biomarkers, namely; IL-1ß, IL-6, and TNF-alpha levels, as well as oxidative stress markers, measured as GSH and MDA. Improved histopathology of the colonic tissue in male New Zealand rabbits also confirmed the superiority of the TXM loaded THs compared to the unformulated drug or the drug free nano-carriers. Our findings highlight the prosperous role of THs in colon targeting and its anti-inflammatory characteristics in guarding against possible NSAIDs-driven exacerbation of colitis.

Keywords: inflammatory bowel disease, trehalose, trehalosomes, colon targeting

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330 Multi-Modal Film Boiling Simulations on Adaptive Octree Grids

Authors: M. Wasy Akhtar

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Multi-modal film boiling simulations are carried out on adaptive octree grids. The liquid-vapor interface is captured using the volume-of-fluid framework adjusted to account for exchanges of mass, momentum, and energy across the interface. Surface tension effects are included using a volumetric source term in the momentum equations. The phase change calculations are conducted based on the exact location and orientation of the interface; however, the source terms are calculated using the mixture variables to be consistent with the one field formulation used to represent the entire fluid domain. The numerical model on octree representation of the computational grid is first verified using test cases including advection tests in severely deforming velocity fields, gravity-based instabilities and bubble growth in uniformly superheated liquid under zero gravity. The model is then used to simulate both single and multi-modal film boiling simulations. The octree grid is dynamically adapted in order to maintain the highest grid resolution on the instability fronts using markers of interface location, volume fraction, and thermal gradients. The method thus provides an efficient platform to simulate fluid instabilities with or without phase change in the presence of body forces like gravity or shear layer instabilities.

Keywords: boiling flows, dynamic octree grids, heat transfer, interface capturing, phase change

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329 Epicardial Fat Necrosis in a Young Female: A Case Report

Authors: Tayyibah Shah Alam, Joe Thomas, Nayantara Shenoy

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Presenting a case that we would like to share, the answer is straight forward but the path taken to get to the diagnosis is where it gets interesting. A 31-year-old lady presented to the Rheumatology Outpatient department with left-sided chest pain associated with left-sided elbow joint pain intensifying over the last 2 days. She had been having a prolonged history of chest pain with minimal intensity since 2016. The pain is intermittent in nature. Aggravated while exerting, lifting heavy weights and lying down. Relieved while sitting. Her physical examination and laboratory tests were within normal limits. An electrocardiogram (ECG) showed normal sinus rhythm and a chest X-ray with no significant abnormality was noted. The primary suspicion was recurrent costochondritis. Cardiac blood inflammatory markers and Echo were normal, ruling out ACS. CT chest and MRI Thorax contrast showed small ill-defined STIR hyperintensity with thin peripheral enhancement in the anterior mediastinum in the left side posterior to the 5th costal cartilage and anterior to the pericardium suggestive of changes in the fat-focal panniculitis. Confirming the diagnosis as Epicardial fat necrosis. She was started on Colchicine and Nonsteroidal anti-inflammatory drugs for 2-3 weeks, following which a repeat CT showed resolution of the lesion and improvement in her. It is often under-recognized or misdiagnosed. CT scan was collectively used to establish the diagnosis. Making the correct diagnosis prospectively alleviates unnecessary testing in favor of conservative management.

Keywords: EFN, panniculitis, unknown etiology, recurrent chest pain

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328 Isolation, Characterization and Myogenic Differentiation of Synovial Mesenchymal Stem Cells

Authors: Fatma Y. Meligy

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Objectives: The objectives of this study aimed to isolate and characterize mesenchymal stem cells (MSCs) derived from synovial membrane. Then to assess the potentiality of myogenic differentiation of these isolated MSCs. Methods: The MSCs were isolated from synovial membrane by digestion method. Three adult rats were used. The 5 -azacytidine was added to the cultured cells for one day. The isolated cells and treated cells are assessed using immunoflouresence, flowcytometry, PCR and real time PCR. Results: The isolated stem cells showed morphological aspect of stem cells they showed strong positivity to CD44 and CD90 in immunoflouresence while in CD34 and CD45 showed negative reaction. The treated cells with 5-azacytidine was shown to have positive reaction for desmin. Flowcytometric analysis showed that synovial MSCs had strong positive percentage for CD44(%98)and CD90 (%97) and low percentage for CD34 & CD45 while the treated cells showed positive percentage for myogenic marker myogenin (85%). As regard the PCR and Real time PCR, the treated cells showed positive reaction to the desmin primer. Conclusion: The adult MSCs were isolated successfully from synovial membrane and characterized with stem cell markers. The isolated cells could be differentiated in vitro into myogenic cells. These differentiated cells could be used in auto-replacement of diseased or traumatized muscle cells as a regenerative therapy for muscle disorders and trauma.

Keywords: mesenchymal stem cells, synovial membrane, myogenic differentiation

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327 Antiplatelet Activity of Nitrated Fatty Acids from Different Food Sources

Authors: Lyanne rodriguez, Eduardo Fuentes, AndrésTrostchansky, Felipe Lagos

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Nutrition is a crucial strategy to combat cardiovascular diseases (CVD), which remain a leading global cause of mortality. Various studies have demonstrated the antiplatelet properties of commonly consumed fruit and vegetable extracts, particularly tomato and bean extracts. Previous research has indicated potent antiplatelet activity in tomato and common bean pomace, attributed to their high fatty acid content (>30%). Notably, fatty acids can undergo nitration during digestion, catalyzed by the reaction of nitrogen dioxide with unsaturated fatty acids. Understanding the mechanisms underlying the formation of nitrated fatty acids from dietary sources is essential to comprehending their antiplatelet action. This research aims to evaluate the formation of nitrated fatty acids (NO₂-FA) from different foods (tomato pulp and common beans). Specifically, tomato pomace and nitrated bean extracts exhibited concentration-dependent antiplatelet effects when platelets were stimulated with TRAP-6 and collagen (1.18±0.04 and 0.7±0.02 mg/mL, respectively). Furthermore, the antiplatelet potential was associated with the modulation of platelet activation markers, as both nitrated extracts suppressed p-selectin expression, CD63 secretion, and fibrinogen modulation. Additionally, a synergistic effect was observed between both nitrated extracts. Our results suggest that NO₂-FA obtained from different food sources has a promising antiplatelet effect for preventing and treating blood clots. This study adds value to these foods in terms of reducing cardiovascular events.

Keywords: foods, nitrated fatty acids, nitration, platelets

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326 The Effect of TiO₂ Nanoparticles on Zebrafish Embryos

Authors: Elena Maria Scalisi

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Currently, photodegradation by nanoparticles (NPs) is a common solution for wastewater treatment. Nanoparticles are efficient for removing organic and inorganic pollutants, heavy metals from wastewater and killing microorganisms through environmentally friendly. In this context, the major representative of photocatalytic technology for industrial wastewater treatment are TiO₂ nanoparticles (TiO₂-NPs). TiO₂-NPs have a strong catalytic activity that depends to their physicochemical properties. Thanks to their small size (between 1-100 nm), nanoparticles occupy less volume, then their surface area increases. The increase in the surface-to-volume ratio results in the increase of the particle surface energy, which improve their reactivity potential. However, these unique properties represent risks to the ecosystems and organisms when unintentionally TiO₂-NPs are release into the environment and absorbed by living organisms. Several studies confirm that there is a high level of interest concerning the safety of TiO₂-NPs in the aquatic environment, furthermore, ecotoxicological tools are useful to correctly evaluate their toxicity. In the current study, we aimed to characterize potential toxic effects of TiO₂-NP suspension to zebrafish during embryo-larval stages to evaluate parameters such as survival rates, malformation, hatching, the overall length of the larvae heartbeat, and biochemical biomarkers that reflect the acute toxicity and sublethal effects of TiO₂-NPs. Zebrafish embryos were exposed to titanium dioxide nanoparticles (TiO₂-NPs at 1mg/L, 2mg/L, and 4mg/L) from fertilization to the free swimming stage (144hpf). Every day, we recorded the toxicological endpoints, moreover, immunohistochemical analysis has been performed at the end of the exposure. In particular, we have evaluate the expression of the following biomarkers: Heat Shock Protein 70 (HSP70), Poly ADP-Ribose Polymerase-1 (PARP-1), Metallothioneins (MTs). Our results have shown that hatch ability, survival, and malformation rate were not affected by TiO₂ NPs at these exposure levels. However, TiO₂-NPs caused an increase of heartbeat and reduction of body length; at the same time, TiO₂-NPs have inducted the production of ROS and the expression of oxidative stress biomarkers HSP70 and PARP-1. Hight positivity for PARP-1 at all concentration tested was observed. As regards MT, positivity was found in the expression of this biomarker in the whole body of the embryo, with the exception of the end of the tail. Metallothioneins (MT) are biomarkers widely used in environmental monitoring programs for aquatic creatures. At the light of our results i.e. no death until the end of the experiment (144hpf), no malformation and expression of the biomarkers mentioned, it is evident that zebrafish larvae with their natural detoxification pathways are able to resist the presence of toxic substances and then they can tolerate the presence of metal concentrations. However, an excessive oxidative state can compromise cell function, therefore the uncontrolled release of nanoparticles into the environment is severe and must be constantly monitored.

Keywords: nanoparticles, embryo zebrafish, HSP70, PARP-1

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325 Amelioration of Over-Expression of bax, Nrf2 and NFК–β in Nano-Sized Titanium Dioxide-Intoxicated Mice by Potent Antioxidants

Authors: Maha Z. Rizk, Sami A. Fattah, Heba M. Darwish, Sanaa A. Ali, Mai O. Kadry

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The increasing use of nanomaterials in consumer and industrial products has aroused global concern regarding their fate in biological systems resulting in demand for parallel risk assessment. The objective of this study is investigating either the effect of individual or combined doses of idebenone, carnosine and vitamin E on amelioration of some biochemical indices of nano sized titanium dioxide (TiO2 NPS) induced metabolic disorders in mice liver. TiO2-NPS was administered in an oral dose of 150 mg/kg for consecutive 14 days followed by oral daily doses of the aforementioned antioxidants for 1 month. TiO2-NPS induced a significant elevation in serum level of ALT and AST, hepatic inflammatory markers (tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6)) and increased the percent of DNA damage which was assessed by COMET assay in addition to the apoptotic marker Caspase-3. Moreover, mRNA gene expression observed by RT-PCR showed a significant overexpression in nuclear factor relation-2 (Nrf2), nuclear factor kappa beta (NF-Kβ) and the apoptotic factor (bax), and a significant down-regulation in the antiapoptotic factor (bcl2) level. In conclusion, idebenone, carnosine and vitamin E ameliorated the deviated parameters with a variable degree with the most pronounced role in alleviating the hazardous effect of TiO2 NPS toxicity following the combination regimen.

Keywords: idebenone, carnosine, vitamin E, TiO2 NPS, caspase-3, NrF2, NF-KB

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324 Effect of Oral Immonoglobulin (IgY) Ingestion on Post Exercise Muscle Soreness and Muscle Damage Markers in Females

Authors: Bert H. Jacobson, Taylor Monaghan, John Sellers

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Intense resistance-type activity generally elicits delayed onset muscle soreness (DOMS) in individuals unaccustomed to such action. DOMS is a combination of contractile tissue microtrauma, osmotic pressure changes, alteration calcium regulation, and inflammation. Elevated muscle-specific enzyme creatine kinase (CK) is a marker of striated muscle damage. Avian immunoglobulin (IgY) mediates inflammation and may thereby reduce post-exercise DOMS. Purpose: The aim of this study was to compare the effect of oral IgY and placebo (Pl) on CK, serum relevels, and perceived pain following induced DOMS. Methods: Healthy college-aged females (N=16) were randomly divided into an experimental group (IgY) and a control group (PL). CK serum levels were recorded followed by 14 days of supplementation of either IgY or Pl at the following doses: days 1-2 =4.5 g, days 3-5 =9.0 g, and days 6-14 =13.5 g. Following the 14 d, lower limb DOMS was induced using two methods of resistance training. After 48 hours, subjects reported for a second blood draw. Results: One-way ANOVA resulted in the IgY group posting significantly less (p < 0.05) serum CK than the PL group. Furthermore, the IgY group experienced significantly less post-test perceived soreness than the Pl group. Conclusion: IgY supplementation lessens muscle CK levels and perceived muscle soreness following exercise, possibly due to an anti-inflammatory effect. It was suggested that IgY may serve as a buffer for DOMS thereby allowing the participant to continue vigorous exercise without discomfort.

Keywords: muscle, soreness, damage, serum

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323 The Effects of Self-Efficacy on Challenge and Threat States

Authors: Nadine Sammy, Mark Wilson, Samuel Vine

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The Theory of Challenge and Threat States in Athletes (TCTSA) states that self-efficacy is an antecedent of challenge and threat. These states result from conscious and unconscious evaluations of situational demands and personal resources and are represented by both cognitive and physiological markers. Challenge is considered a more adaptive stress response as it is associated with a more efficient cardiovascular profile, as well as better performance and attention effects compared with threat. Self-efficacy is proposed to influence challenge/threat because an individual’s belief that they have the skills necessary to execute the courses of action required to succeed contributes to a perception that they can cope with the demands of the situation. This study experimentally examined the effects of self-efficacy on cardiovascular responses (challenge and threat), demand and resource evaluations, performance and attention under pressurised conditions. Forty-five university students were randomly assigned to either a control (n=15), low self-efficacy (n=15) or high self-efficacy (n=15) group and completed baseline and pressurised golf putting tasks. Self-efficacy was manipulated using false feedback adapted from previous studies. Measures of self-efficacy, cardiovascular reactivity, demand and resource evaluations, task performance and attention were recorded. The high self-efficacy group displayed more favourable cardiovascular reactivity, indicative of a challenge state, compared with the low self-efficacy group. The former group also reported high resource evaluations, but no task performance or attention effects were detected. These findings demonstrate that levels of self-efficacy influence cardiovascular reactivity and perceptions of resources under pressurised conditions.

Keywords: cardiovascular, challenge, performance, threat

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322 Durian Marker Kit for Durian (Durio zibethinus Murr.) Identity

Authors: Emma K. Sales

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Durian is the flagship fruit of Mindanao and there is an abundance of several cultivars with many confusing identities/ names. The project was conducted to develop procedure for reliable and rapid detection and sorting of durian planting materials. Moreover, it is also aimed to establish specific genetic or DNA markers for routine testing and authentication of durian cultivars in question. The project developed molecular procedures for routine testing. SSR primers were also screened and identified for their utility in discriminating durian cultivars collected. Results of the study showed the following accomplishments; 1. Twenty (29) SSR primers were selected and identified based on their ability to discriminate durian cultivars, 2. Optimized and established standard procedure for identification and authentication of Durian cultivars 3. Genetic profile of durian is now available at Biotech Unit. Our results demonstrate the relevance of using molecular techniques in evaluating and identifying durian clones. The most polymorphic primers tested in this study could be useful tools for detecting variation even at the early stage of the plant especially for commercial purposes. The process developed combines the efficiency of the microsatellites development process with the optimization of non-radioactive detection process resulting in a user-friendly protocol that can be performed in two (2) weeks and easily incorporated into laboratories about to start microsatellite development projects. This can be of great importance to extend microsatellite analyses to other crop species where minimal genetic information is currently available. With this, the University can now be a service laboratory for routine testing and authentication of durian clones.

Keywords: DNA, SSR analysis, genotype, genetic diversity, cultivars

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321 Introgressive Hybridisation between Two Widespread Sharks in the East Pacific Region

Authors: Diana A. Pazmino, Lynne vanHerwerden, Colin A. Simpfendorfer, Claudia Junge, Stephen C. Donnellan, Mauricio Hoyos-Padilla, Clinton A. J. Duffy, Charlie Huveneers, Bronwyn Gillanders, Paul A. Butcher, Gregory E. Maes

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With just a handful of documented cases of hybridisation in cartilaginous fishes, shark hybridisation remains poorly investigated. Small amounts of admixture have been detected between Galapagos (Carcharhinus galapagensis) and dusky (Carcharhinus obscurus) sharks previously, generating a hypothesis of ongoing hybridisation. We sampled a large number of individuals from areas where both species co-occur (contact zones) across the Pacific Ocean and used both mitochondrial and nuclear-encoded SNPs to examine genetic admixture and introgression between the two species. Using empirical, analytical approaches and simulations, we first developed a set of 1,873 highly informative and reliable diagnostic SNPs for these two species to evaluate the degree of admixture between them. Overall, results indicate a high discriminatory power of nuclear SNPs (FST=0.47, p < 0.05) between the two species, unlike mitochondrial DNA (ΦST = 0.00 p > 0.05), which failed to differentiate between these species. We identified four hybrid individuals (~1%) and detected bi-directional introgression between C. galapagensis and C. obscurus in the Gulf of California along the eastern Pacific coast of the Americas. We emphasize the importance of including a combination of mtDNA and diagnostic nuclear markers to properly assess species identification, detect patterns of hybridisation, and better inform management and conservation of these sharks, especially given the morphological similarities within the genus Carcharhinus.

Keywords: elasmobranchs, single nucleotide polymorphisms, hybridisation, introgression, misidentification

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320 L-Carnitine Supplementation and Exercise-Induced Muscle Damage

Authors: B. Nakhostin-Roohi, F. Khoshkhahesh, KH. Parandak, R. Ramazanzadeh

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Introduction: The protective effect of antioxidants in diminishing the post-exercise rise of serum CK and LDH in individuals trained for competitive sports has come to light in recent years. This study was conducted to assess the effect of Two-week L-carnitine supplementation on exercise-induced muscle damage, as well as antioxidant capacity after a bout of strenuous exercise in active healthy young men. Methodology: Twenty active healthy men volunteered for this study. Participants were randomized in a double-blind placebo-controlled fashion into two groups: L-carnitine (C group; n = 10) and placebo group (P group; n = 10). The participants took supplementation (2000 mg L-carnitine) or placebo (2000 mg lactose) daily for 2weeks before the main trial. Then, participants ran 14 km. Blood samples were taken before supplementation, before exercise, immediately, 2h and 24h after exercise. Creatine kinase (CK), and lactate dehydrogenase (LDH), and total antioxidant capacity (TAC) were measured. Results: Serum CK and LDH significantly increased after exercise in both groups (p < 0.05). Serum LDH was significantly lower in C group than P group 2h and 24h after exercise (p < 0.05). Furthermore, CK was significantly lower in C group compared with P group just 24h after exercise (p < 0.05). Plasma TAC increased significantly 14 days after supplementation and 24h after exercise in C group compared with P group (p < 0.05). Discussion and conclusion: These results suggest two-week daily oral supplementation of L-carnitine has been able to promote antioxidant capacity before and after exercise and decrease muscle damage markers through possibly inhibition of exercise-induced oxidative stress.

Keywords: L-carnitine, muscle damage, creatine kinase, Lactate dehydrogenase

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319 Isolated Iterating Fractal Independently Corresponds with Light and Foundational Quantum Problems

Authors: Blair D. Macdonald

Abstract:

After nearly one hundred years of its origin, foundational quantum mechanics remains one of the greatest unexplained mysteries in physicists today. Within this time, chaos theory and its geometry, the fractal, has developed. In this paper, the propagation behaviour with an iteration of a simple fractal, the Koch Snowflake, was described and analysed. From an arbitrary observation point within the fractal set, the fractal propagates forward by oscillation—the focus of this study and retrospectively behind by exponential growth from a point beginning. It propagates a potentially infinite exponential oscillating sinusoidal wave of discrete triangle bits sharing many characteristics of light and quantum entities. The model's wave speed is potentially constant, offering insights into the perception and a direction of time where, to an observer, when travelling at the frontier of propagation, time may slow to a stop. In isolation, the fractal is a superposition of component bits where position and scale present a problem of location. In reality, this problem is experienced within fractal landscapes or fields where 'position' is only 'known' by the addition of information or markers. The quantum' measurement problem', 'uncertainty principle,' 'entanglement,' and the classical-quantum interface are addressed; these are a problem of scale invariance associated with isolated fractality. Dual forward and retrospective perspectives of the fractal model offer the opportunity for unification between quantum mechanics and cosmological mathematics, observations, and conjectures. Quantum and cosmological problems may be different aspects of the one fractal geometry.

Keywords: measurement problem, observer, entanglement, unification

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318 Evaluation of Immunostimulant Potential of Proteoliposomes Derived from Vibrio anguillarum Administered by Immersion in Zebrafish (Danio rerio)

Authors: M. Caruffo, P. Navarrete, C. G. Feijoo, L. Sáenz

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Disease prevention through the use of vaccines has been crucial to achieve the current level of production in the salmon industry. However, vaccines have been developed based largely on inactivated bacterial formulations, using the whole pathogen. These formulations have demonstrated excellent efficacy against extracellular bacterial pathogens. However diseases with the greatest economic impacts correspond to intracellular bacterial and viral pathogens, vaccines based on these types of agents have shown a discrete effectiveness. It is for these reasons that the development of subunit vaccines based on defined antigens offers a promising solution. The main problem is that subunit vaccines offer a low immunogenicity, since they lack immunostimulatory elements, so that the development of new adjuvants platforms becomes an important challenge for this type of formulations. We evaluate the effect of a formulation based on proteoliposomes of Vibrio anguillarum administered by immersion as a new adjuvant strategy, allowing efficient stimulation of the innate immune system. Proteoliposomes physicochemical properties were evaluated in its ability to produce an inflammatory process. Using zebrafish (Danio rerio) larvae as a model species and the transgenic line (Tg(mpx: GFP)i114) allowed us to track the neutrophil migration in real time. Additionally we evaluated the gene expression of some molecular markers involved in the development of the innate immune response characterizing the adjuvant capacity of the formulation.

Keywords: adjuvants, vaccine development, zebrafish, innate immunity

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317 Anthocyanins as Markers of Enhanced Plant Defence in Maize (Zea Mays L.) Exposed to Copper Stress

Authors: Fadime Eryılmaz Pehlivan

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Anthocyanins are important plant pigments having roles in many physiological and ecological functions; that are controlled by numerous regulatory factors. The accumulation of anthocyanins in Z. mays cause the plants stems to exhibit red coloration when encountering gradually increasing copper treatments (1, 5, and 10 mM of Cu in a period of 5 days) on maize seedlings. Stress injury was measured in terms of chlorophyll (a and b), carotenoid and anthocyanin contents, malondialdehyde (MDA), hydrogen peroxide (H2O2). Carotenoid and anthocyanin contents dramatically increased by increasing concentrations of Cu stress. MDA and H2O2 levels were found to significantly increase at high Cu treatments (5 and 10 mM of Cu). Chlorophyll content was observed to be highest at 1 mM Cu and then decreased at 5 and 10 mM of Cu. In addition, significant increases were determined in the activities of catalase (CAT), superoxide dismutase (SOD), glutathione reductase (GR) and ascorbate peroxidase (APX) under high Cu concentrations, while glutathione S-transferase (GST) and peroxidase (POX) activities showed no change. Treatments above 5 and 10 mM of Cu triggered copper stress in maize seedlings. The results of this study provide evidence that maize seedlings represent a high tolerance to gradually increasing copper treatments. Improved copper tolerance may relate to high anthocyanin, and carotenoid content besides antioxidant enzyme activity may improve the metal chelating ability of anthocyanin pigments. Data presented in this study may also contribute to a better understanding of phytoremediation studies in maize exposed to high copper contenting soils.

Keywords: anthocyanin, copper, maize , antioxidant

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316 Effect of Visnagin on Altered Steroidogenesis and Spermatogenesis, and Testicular Injury Induced by the Heavy Metal Lead

Authors: Saleh N. Maodaa

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Background: Lead (Pb) is an environmental pollutant causing serious health problems, including impairment of reproduction. Visnagin (VIS) is a furanochromone with promising antioxidant and anti-inflammatory effects; however, its protective efficacy against Pb toxicity has not been investigated. Objective: This study evaluated the protective effect of VIS on Pb reproductive toxicity, impaired steroidogenesis and spermatogenesis, oxidative stress and inflammation. Methods: Rats received VIS (30 or 60 mg/kg) and 50 mg/kg lead acetate for 3 weeks, and blood and testes samples were collected. Results: Pb intoxication impaired the pituitary-testicular axis (PTA), manifested by the decreased serum levels of gonadotropins and testosterone. Pb decreased sperm count, motility and viability, increased sperm abnormalities, and downregulated the steroidogenesis markers StAR, CYP17A1, 3β-HSD and 17β-HSD in the testis of rats. VIS significantly increased serum gonadotropins and testosterone, alleviated sperm parameters and upregulated steroidogenesis. In addition, VIS decreased pro-inflammatory cytokines, testicular lipid peroxidation and DNA fragmentation, downregulated Bax, and enhanced antioxidants and Bcl-2 Conclusion: These results demonstrate the protective effect of VIS against Pb reproductive toxicity in rats. VIS improved serum gonadotropins and testosterone, enhanced steroidogenesis and spermatogenesis, and attenuated oxidative injury, inflammation and apoptosis. Therefore, VIS is a promising candidate for the protection against Pb-induced reproduction impairment.

Keywords: pituitary-gonadal axis, cytokines, DNA damage, apoptosis

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315 Comparison of Methods for the Synthesis of Eu+++, Tb+++, and Tm+++ Doped Y2O3 Nanophosphors by Sol-Gel and Hydrothermal Methods for Bioconjugation

Authors: Ravindra P. Singh, Drupad Ram, Dinesh K. Gupta

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Rare earth ions doped metal oxides are a class of luminescent materials which have been proved to be excellent for applications in field emission displays and cathode ray tubes, plasma display panels. Under UV irradiation Eu+++ doped Y2O3 is a red phosphor and Tb+++ doped Y 2O3 is a green phosphor. It is possible that, due to their high quantum efficiency, they might serve as improved luminescent markers for identification of biomolecules, as already reported for CdSe and CdSe/ZnS nanocrystals. However, for any biological applications these particle powders must be suspended in water while retaining their phosphorescence. We hereby report synthesis and characterization of Eu+++ and Tb+++ doped yttrium oxide nanoparticles by sol-gel and hydrothermal processes. Eu+++ and Tb+++ doped Y2O3 nanoparticles have been synthesized by hydrothermal process using yttrium oxo isopropoxide [Y5O(OPri)13] (crystallized twice) and it’s acetyl acetone modified product [Y(O)(acac)] as precursors. Generally the sol-gel derived metal oxides are required to be annealed to the temperature ranging from 400°C-800°C in order to develop crystalline phases. However, this annealing also results in the development of aggregates which are undesirable for bio-conjugation experiments. In the hydrothermal process, we have achieved crystallinity of the nanoparticles at 300°C and the development of crystalline phases has been found to be proportional to the time of heating of the reactor. The average particle sizes as calculated from XRD were found to be 28 nm, 32 nm, and 34 nm by hydrothermal process. The particles were successfully suspended in chloroform in the presence of trioctyl phosphene oxide and TEM investigations showed the presence of single particles along with agglomerates.

Keywords: nanophosphors, Y2O3:Eu+3, Y2O3:Tb+3, sol-gel, hydrothermal method, TEM, XRD

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314 Polyphytopharmaca Improving Asthma Control Test Value, Biomarker (Eosinophils and Malondialdehyde): Quasi Experimental Test in Patients with Asthma

Authors: Andri Andri, Susanthy Djajalaksana, Iin Noor Chozin

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Background: Despite advances in asthma therapies, a proportion of patients with asthma continue to have difficulty in gaining adequate asthma control. Complex immunological mechanisms and oxidative stress affect this condition, including the role of malondialdehyde (MDA) as a marker of inflammation. This research aimed to determine the effect of polyphytopharmaca administration on the value of asthma control test (ACT), blood eosinophils level and markers of MDA serum inflammation in patients with asthma. Method: Quasi experimental approach was conducted toward 15 stable asthma patients who were not fully controlled in outpatient pulmonary clinic, Public Hospital of Dr. Saiful Anwar Malang. Assessments of ACT values, eosinophil levels, and serum MDA levels were carried out before and after administration of polyphytopharmaca which contained a combination of 100 mg Nigella sativa extract, Kleinhovia hospita 100 mg, Curcuma xanthorrhiza 75 mg, and Ophiocephalus striatus 100 mg, three times daily with two capsules for 12 weeks. The ACT value was determined by the researcher by asking the patient directly, blood eosinophil levels were calculated by analyzing blood type counts, and serum MDA levels were detected by the qPCR method. Result: There was a significant enhancement of ACT value (18.07 ± 2.57 to 22.06 ± 1.83, p = 0.001) (from 60% uncontrolled ACT to 93.3% controlled ACT), a significant decrease in blood eosinophils levels (653.15 ± 276.15 pg/mL to 460.66 ± 202.04 pg/mL, p = 0.038), and decreased serum MDA levels (109.64 ± 53.77 ng / ml to 78.68 ± 64.92 ng/ml, p = 0.156). Conclusion: Administration of polyphytopharmaca can increase ACT value, decrease blood eosinophils levels and reduce MDA serum in stable asthma patients who are not fully controlled.

Keywords: asthma control test, eosinophils levels, malondialdehyde, polyphytopharmaca

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313 The Improved Therapeutic Effect of Trans-Cinnamaldehyde on Adipose-Derived Stem Cells without Chemical Induction

Authors: Karthyayani Rajamani, Yi-Chun Lin, Tung-Chou Wen, Jeanne Hsieh, Yi-Maun Subeq, Jen-Wei Liu, Po-Cheng Lin, Horng-Jyh Harn, Shinn-Zong Lin, Tzyy-Wen Chiou

Abstract:

Assuring cell quality is an essential parameter for the success of stem cell therapy, utilization of various components to improve this potential has been the primary goal of stem cell research. The aim of this study was not only to demonstrate the capacity of trans-cinnamaldehyde (TC) to reverse stress-induced senescence but also improve the therapeutic abilities of stem cells. Because of the availability and the promising application potential in regenerative medicine, adipose-derived stem cells (ADSCs) were chosen for the study. We found that H2O2 treatment resulted in the expression of senescence characteristics in the ADSCs, including decreased proliferation rate, increased senescence-associated- β-galactosidase (SA-β-gal) activity, decreased SIRT1 (silent mating type information regulation 2 homologs) expression and decreased telomerase activity. However, TC treatment was sufficient to rescue or reduce the effects of H2O2 induction, ultimately leading to an increased proliferation rate, a decrease in the percentage of SA-β-gal positive cells, upregulation of SIRT1 expression, and increased telomerase activity of the senescent ADSCs at the cellular level. Further recently it was observed that the ADSCs were treated with TC without induction of senescence, all the before said positives were observed. Moreover, a chemically induced liver fibrosis animal model was used to evaluate the functionality of these rescued cells in vivo. Liver dysfunction was established by injecting 200 mg/kg thioacetamide (TAA) intraperitoneally into Wistar rats every third day for 60 days. The experimental rats were separated into groups; normal group (rats without TAA induction), sham group (without ADSC transplantation), positive control group (transplanted with normal ADSCs); H2O2 group (transplanted with H2O2 -induced senescent ADSCs), H2O2+TC group (transplanted with ADSCs pretreated with H2O2 and then further treated with TC) and TC group (ADSC treated with TC without H2O2 treatment). In the transplantation group, 1 × 106 human ADSCs were introduced into each rat via direct liver injection. Based on the biochemical analysis and immunohistochemical staining results, it was determined that the therapeutic effects on liver fibrosis by the induced senescent ADSCs (H2O2 group) were not as significant as those exerted by the normal ADSCs (the positive control group). However, the H2O2+TC group showed significant reversal of liver damage when compared to the H2O2 group 1 week post-transplantation. Further ADSCs without H2O2 treatment but with just TC treatment performed much better than all the groups. These data confirmed that the TC treatment had the potential to improve the therapeutic effect of ADSCs. It is therefore suggested that TC has potential applications in maintaining stem cell quality and could possibly aid in the treatment of senescence-related disorders.

Keywords: senescence, SIRT1, adipose derived stem cells, liver fibrosis

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312 The Effects of Three Months of HIIT on Plasma Adiponectin on Overweight College Men

Authors: M. J. Pourvaghar, M. E. Bahram, M. Sayyah, Sh. Khoshemehry

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Adiponectin is a cytokine secreted by the adipose tissue that functions as an anti-inflammatory, antiathrogenic and anti-diabetic substance. Its density is inversely correlated with body mass index. The purpose of this research was to examine the effect of 12 weeks of high intensity interval training (HIIT) with the level of serum adiponectin and some selected adiposity markers in overweight and fat college students. This was a clinical research in which 24 students with BMI between 25 kg/m2 to 30 kg/m2. The sample was purposefully selected and then randomly assigned into two groups of experimental (age =22.7±1.5 yr.; weight = 85.8±3.18 kg and height =178.7±3.29 cm) and control (age =23.1±1.1 yr.; weight = 79.1±2.4 kg and height =181.3±4.6 cm), respectively. The experimental group participated in an aerobic exercise program for 12 weeks, three sessions per weeks at a high intensity between 85% to 95% of maximum heart rate (considering the over load principle). Prior and after the termination of exercise protocol, the level of serum adiponectin, BMI, waist to hip ratio, and body fat percentages were calculated. The data were analyzed by using SPSS: PC 16.0 and statistical procedure such as ANCOVA, was used. The results indicated that 12 weeks of intensive interval training led to the increase of serum adiponectin level and decrease of body weight, body fat percent, body mass index and waist to hip ratio (P < 0.05). Based on the results of this research, it may be concluded that participation in intensive interval training for 12 weeks is a non-invasive treatment to increase the adiponectin level while decreasing some of the anthropometric indices associated with obesity or being overweight.

Keywords: adiponectin, cardiovascular, interval, overweight, training

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311 Down-Regulated Gene Expression of GKN1 and GKN2 as Diagnostic Markers for Gastric Cancer

Authors: Amer A. Hasan, Mehri Igci, Ersin Borazan, Rozhgar A. Khailany, Emine Bayraktar, Ahmet Arslan

Abstract:

Gastric cancer (GC) has high morbidity and fatality rate in various countries and is still one of the most frequent and deadly diseases. Novel mitogenic and motogenic Gastrokine1 (GKN1) and Gastrokine 2 (GKN2) genes that are highly expressed in the normal stomach epithelium and plays an important role in maintaining the integrity and homeostasis of stomach mucosal epithelial cells. Significant loss of copy number and mRNA transcript of GKN1 and GKN2 gene expression were frequently observed in all types of gastric cancer. In this study, 47 paired samples that were grouped according to the types of gastric cancer and the clinical characteristics of the patients, including gender and average of age were investigated with gene expression analysis and mutation screening by monetering RT-PCR, SSCP and nucleotide sequencing techniques. Both GKN1 and GKN2 genes were observed significantly reduced found by (Wilcoxon signed rank test; p<0.05). As a result of gene screening, no mutation (no different genotype) was detected. It is considered that gene mutations are not the cause of inactivation of gastrokines. In conclusion, the mRNA expression level of GKN1 and GKN2 genes statistically was decreased regardless the gender, age or cancer type of patients. Reduced of gastrokine genes seems to occur at the initial steps of cancer development. In order to understand the investigation between gastric cancer and diagnostic biomarker; further analysis is necessary.

Keywords: gastric cancer, diagnostic biomarker, nucleotide sequencing, semi-quantitative RT-PCR

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310 Co-Administration Effects of Conjugated Linoleic Acid and L-Carnitine on Weight Gain and Biochemical Profile in Diet Induced Obese Rats

Authors: Maryam Nazari, Majid Karandish, Alihossein Saberi

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Obesity as a global health challenge motivates pharmaceutical industries to produce anti-obesity drugs. However, effectiveness of these agents is remained unclear. Because of popularity of dietary supplements, the aim of this study was tp investigate the effects of Conjugated Linoleic Acid (CLA) and L-carnitine (LC) on serum glucose, triglyceride, cholesterol and weight changes in diet induced obese rats. 48 male Wistar rats were randomly divided into two groups: Normal fat diet (n=8), and High fat diet (HFD) (n=32). After eight weeks, the second group which was maintained on HFD until the end of study, was subdivided into four categories: a) 500 mg Corn Oil (as control group), b) 500 mg CLA, c) 200 mg LC, d) 500 mg CLA+ 200 mg LC.All doses are planned per kg body weights, which were administered by oral gavage for four weeks. Body weights were measured and recorded weekly by means of a digital scale. At the end of the study, blood samples were collected for biochemical markers measurement. SPSS Version 16 was used for statistical analysis. At the end of 8th week, a significant difference in weight was observed between HFD and NFD group. After 12 weeks, LC significantly reduced weight gain by 4.2%. Trend of weight gain in CLA and CLA+LC groups was insignificantly decelerated. CLA+LC reduced triglyceride level significantly, but just CLA had significant influence on total cholesterol and insignificant decreasing effect on FBS. Our results showed that an obesogenic diet in a relative short time led to obesity and dyslipidemia which can be modified by LC and CLA to some extent.

Keywords: conjugated linoleic acid, high fat diet, L-Carnitine, obesity

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309 Liquid Biopsy and Screening Biomarkers in Glioma Grading

Authors: Abdullah Abdu Qaseem Shamsan

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Background: Gliomas represent the most frequent, heterogeneous group of tumors arising from glial cells, characterized by difficult monitoring, poor prognosis, and fatality. Tissue biopsy is an established procedure for tumor cell sampling that aids diagnosis, tumor grading, and prediction of prognosis. We studied and compared the levels of liquid biopsy markers in patients with different grades of glioma. Also, it tried to establish the potential association between glioma and specific blood groups antigen. Result: 78 patients were identified, among whom maximum percentage with glioblastoma possessed blood group O+ (53.8%). The second highest frequency had blood group A+ (20.4%), followed by B+ (9.0%) and A- (5.1%), and least with O-. Liquid biopsy biomarkers comprised of ALT, LDH, lymphocytes, Urea, Alkaline phosphatase, AST Neutrophils, and CRP. The levels of all the components increased significantly with the severity of glioma, with maximum levels seen in glioblastoma (grade IV), followed by grade III and grade II respectively. Conclusion: Gliomas possess significant clinical challenges due to their progression with heterogeneous nature and aggressive behavior. Liquid biopsy is a non-invasive approach which aids to establish the status of the patient and determine the tumor grade, therefore may show diagnostic and prognostic utility. Additionally, our study provides evidence to demonstrate the role of ABO blood group antigens in the development of glioma. However, future clinical research on liquid biopsy will improve the sensitivity and specificity of these tests and validate their clinical usefulness to guide treatment approaches.

Keywords: GBM: glioblastoma multiforme, CT: computed tomography, MRI: magnetic resonance imaging, ctRNA: circulating tumor RNA

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308 Establishing Sequence Stratigraphic Framework and Hydrocarbon Potential of the Late Cretaceous Strata: A Case Study from Central Indus Basin, Pakistan

Authors: Bilal Wadood, Suleman Khan, Sajjad Ahmed

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The Late Cretaceous strata (Mughal Kot Formation) exposed in Central Indus Basin, Pakistan is evaluated for establishing sequence stratigraphic framework and potential of hydrocarbon accumulation. The petrographic studies and SEM analysis were carried out to infer the hydrocarbon potential of the rock unit. The petrographic details disclosed 4 microfacies including Pelagic Mudstone, OrbitoidalWackestone, Quartz Arenite, and Quartz Wacke. The lowermost part of the rock unit consists of OrbitoidalWackestone which shows deposition in the middle shelf environment. The Quartz Arenite and Quartz Wacke suggest deposition on the deep slope settings while the Pelagic Mudstone microfacies point toward deposition in the distal deep marine settings. Based on the facies stacking patterns and cyclicity in the chronostratigraphic context, the strata is divided into two 3rd order cycles. One complete sequence i.e Transgressive system tract (TST), Highstand system tract (HST) and Lowstand system tract (LST) are again replaced by another Transgressive system tract and Highstant system tract with no markers of sequence boundary. The LST sands are sandwiched between TST and HST shales but no potential porosity/permeability values have been determined. Microfacies and SEM studies revealed very fewer chances for hydrocarbon accumulation and overall reservoir potential is characterized as low.

Keywords: cycle, deposition, microfacies, reservoir

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