Search results for: spawning induction
219 Effect of Ocimum americanum Water Extract on Antioxidant System in Rat
Authors: Pornrut Rabintossaporn, Suphaket Saenthaweesuk, Amornnat Thuppia, Nuntiya Somparn
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Several dietary and herbal plants have been shown to possess cytoprotective and antioxidant effects with various mechanisms of action. The aim of this study was to determine the antioxidant effects and its mechanism of aqueous leaves extract of Ocimum americanum (OA), commonly known as American basil or 'hoary basil', in rat. The extract was screened for its phytochemical contents and antioxidant activity in vitro. Moreover, the extract was studied in rats to evaluate its effects in vivo. Rats were orally administered with the extract at the dose of 100, 200 and 400 mg/kg for 28 days. Phytochemical screening of plant extracts revealed the presence of alkaloid, cardiac glycosides, tannin and steroid compounds. The extract contained phenolic compounds 36.91 ± 0.66 mg of gallic acid equivalents per gram OA extract. The free radical scavenging activity assessed by DPPH assay gave IC50 of 41.27 ± 1.86 µg/mL, which is relatively lower than that of BHT with IC50 of 12.34 ± 1.14µg/mL. In the animals, the extract was well tolerated by the animals throughout the 28 days of study as shown by normal serum levels AST, ALP, ALT, BUN and Cr as well as normal histology of liver and pancreatic and kidney tissue. The protein expression of antioxidant enzymes, γ-glutamylcysteine ligase (γ-GCL) in liver was significantly increased compared with normal control. Consistent with the induction of γ-GCL protein expression significantly reduction of serum oxidative stress marker malondialdehyde (MDA) was found in rat treated with OA extract compared with control. Taken together, this study provides evidence that Ocimum americanum exhibits direct antioxidant properties and can induce cytoprotective enzyme in vivo.Keywords: antioxidant, γ-glutamylcysteine ligase, MDA, Ocimum americanum
Procedia PDF Downloads 243218 The Generation of Insulin Producing Cells from Human Mesenchymal Stem Cells by miR-375 and Anti-miR-9
Authors: Arefeh Jafarian, Mohammad Taghikani, Saied Abroun, Amir Allahverdi, Masoud Soleimani
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Introduction: The miRNAs have key roles in control of pancreatic islet development and insulin secretion. In this regards, current study investigated the pancreatic differentiation of human bone marrow mesenchymal stem cells (hBM-MSCs) by up-regulation of miR-375 and down-regulation of miR-9 by lentiviruses containing miR-375 and anti-miR-9. Findings: After 21 days of induction, islet-like clusters containing insulin producing cells (IPCs) were confirmed by dithizone (DTZ) staining. The IPCs and β cell specific related genes and proteins were detected using qRT-PCR and immunofluorescence on days 7, 14 and 21 of differentiation. Glucose challenge test was performed at different concentrations of glucose as well as extracellular and intracellular insulin and C-peptide were assayed using ELISA kit. In derived IPCs by miR-375 alone are capable to express insulin and other endocrine specific transcription factors, the cells lack the machinery to respond to glucose. The differentiated hMSCs by miR-375 and anti-miR-9 lentiviruses could secrete insulin and c-peptide in a glucose-regulated manner. Conclusion: It was found that over-expression of miR-375 led to a reduction in levels of Mtpn protein in derived IPCs, while treatment with anti-miR-9 following miR-375 over-expression had synergistic effects on MSCs differentiation and insulin secretion in a glucose-regulated manner. The researchers reported that silencing of miR-9 increased OC-2 protein in IPCs that may contribute to the observed glucose-regulated insulin secretion. These findings highlight miRNAs functions in stem cells differentiation and suggest that they could be used as therapeutic tools for gene-based therapy in diabetes mellitus.Keywords: diabetes, differentiation, MSCs, insulin producing cells, miR-375, miR-9
Procedia PDF Downloads 317217 Positive effect of Cu2+ and Ca2+ on the Thermostability of Bambara Groundnut Peroxidase A6, and its Catalytic Efficiency Toward the Oxidation of 3,3,5,5 -Tetramethyl Benzidine
Authors: Yves Mann Elate Lea Mbassi, Marie Solange Evehe Bebandoue, Wilfred Fon Mbacham
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Improving the catalytic performance of enzymes has been a long-standing theme of analytical biochemistry research. Induction of peroxidase activity by metals is a common reaction in higher plants. We thought that this increase in peroxidase activity may be due, on the one hand, to the stimulation of the gene expression of these enzymes but also to a modification of their chemical reactivity following the binding of some metal ions on their active site. We tested the effect of some metal salts (MgCl₂, MnCl₂, ZnCl₂, CaCl₂ and CuSO₄) on the activity and thermostability of peroxidase A6, a thermostable peroxidase that we discovered and purified in a previous study. The chromogenic substrate used was 3,3′,5,5′-tetramethylbenzidine. Of all the metals tested for their effect on A6, only magnesium and copper had a significant effect on the activity of the enzyme at room temperature. The Mann-Whitney test shows a slight inhibitory effect of activity by the magnesium salt (P = 0.043), while the activity of the enzyme is 5 times higher in the presence of the copper salt (P = 0.002). Moreover, the thermostability of peroxidase A6 is increased when calcium and copper salts are present. The activity in the presence of CaCl₂ is 8 times higher than the residual activity of the enzyme alone after incubation at 80°C for 10 min and 35 times higher in the presence of CuSO4 under the same conditions. In addition, manganese and zinc salts slightly reduce the thermostability of the enzyme. The activity and structural stability of peroxidase A6 can clearly be activated by Cu₂+, which therefore enhance the oxidation of 3,3′,5,5′-tetramethylbenzidine, which was used in this study as a chromogenic substrate. Ca₂+ likely has a more stabilizing function for the catalytic site.Keywords: peroxidase activity, copper ions, calcium ions, thermostability
Procedia PDF Downloads 73216 High Temperature Deformation Behavior of Al0.2CoCrFeNiMo0.5 High Entropy alloy
Authors: Yasam Palguna, Rajesh Korla
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The efficiency of thermally operated systems can be improved by increasing the operating temperature, thereby decreasing the fuel consumption and carbon footprint. Hence, there is a continuous need for replacing the existing materials with new alloys with higher temperature working capabilities. During the last decade, multi principal element alloys, commonly known as high entropy alloys are getting more attention because of their superior high temperature strength along with good high temperature corrosion and oxidation resistance, The present work focused on the microstructure and high temperature tensile behavior of Al0.2CoCrFeNiMo0.5 high entropy alloy (HEA). Wrought Al0.2CoCrFeNiMo0.5 high entropy alloy, produced by vacuum induction melting followed by thermomechanical processing, is tested in the temperature range of 200 to 900oC. It is exhibiting very good resistance to softening with increasing temperature up to 700oC, and thereafter there is a rapid decrease in the strength, especially beyond 800oC, which may be due to simultaneous occurrence of recrystallization and precipitate coarsening. Further, it is exhibiting superplastic kind of behavior with a uniform elongation of ~ 275 % at 900 oC temperature and 1 x 10-3 s-1 strain rate, which may be due to the presence of fine stable equi-axed grains. Strain rate sensitivity of 0.3 was observed, suggesting that solute drag dislocation glide might be the active mechanism during superplastic kind of deformation. Post deformation microstructure suggesting that cavitation at the sigma phase-matrix interface is the failure mechanism during high temperature deformation. Finally, high temperature properties of the present alloy will be compared with the contemporary high temperature materials such as ferritic, austenitic steels, and superalloys.Keywords: high entropy alloy, high temperature deformation, super plasticity, post-deformation microstructures
Procedia PDF Downloads 164215 Antioxidant Responses and Malondialdehyde Levels in African Cat Fish (Clarias gariepinus) from Eleyele River in Nigeria
Authors: Oluwatosin Adetola Arojojoye, Olajumoke Olufunlayo Alao, Philip Odigili
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This study investigated the extent of pollution in Eleyele River in Oyo State, Nigeria by investigating the antioxidant status and malondialdehyde levels (index of lipid peroxidation) in the organs of African Catfish, Clarias gariepinus from the river. Clarias gariepinus weighing between 250g-400g were collected from Eleyele River (a suspected polluted river) and Clarias gariepinus from a clean fish farm (Durantee fisheries) were used as the control. Levels of malondialdehyde, glutathione concentration (GSH) and activities of antioxidant enzymes - superoxide dismutase, catalase and glutathione-S-transferase (GST) were evaluated in the post-mitochondrial fractions of the liver, kidney and gills of the fishes. From the results, there were increases in malondialdehyde level and GSH concentration in the liver, kidney and gills of Clarias gariepinus from Eleyele River when compared with control. Glutathione-S-transferase activity was induced in the liver and kidney of Clarias gariepinus from Eleyele River when compared with control. However, the activity of this enzyme was depleted in the gills of fishes from Eleyele River compared with control. Also there was an induction in SOD activity in the liver of Clarias gariepinus from Eleyele River when compared with control but there was a decrease in the activity of this enzyme in the kidney and gills of fishes from Eleyele River compared with control. Increase in lipid peroxidation and alterations in antioxidant system in Clarias gariepinus from Eleyele River show that the fishes were under oxidative stress. These suggest that the river is polluted probably as a result of industrial, domestic and agricultural wastes frequently discharged into the river. This could pose serious health risks to consumers of water and aquatic organisms from the river.Keywords: antioxidant, lipid peroxidation, Clarias gariepinus, Eleyele River
Procedia PDF Downloads 529214 Rebamipide Retards CCL4 Induced Hepatic Fibrosis: A Role of PGE2
Authors: Alaa E. El-sisi, Sherin Zakaria
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Rebamipide is an antiulcer drug with unique properties such as anti-inflammatory action. It induces endogenous prostaglandin e2 (PGE2). PGE2 is considered as a potent physiological suppressor of liver fibrosis. Aim of study: This study investigated the effect of rebamipide on hepatic fibrosis. Material and Method: Hepatic fibrosis was induced by intraperitoneal injections (IP) injection of CCl4 (0.45 mL/kg) in corn oil 1:5 twice a week for 4 weeks. Rats were divided into four groups as follow: Group 1 treated with CCL4 only, group 2 and 3 treated with CCL4 and rebamipide 60 mg/kg/day (group2) or 100 mg/kg/day (group3), and the fourth group was considered as control group and treated with vehicles. ALT, AST, and Bilirubin were assayed in serum. Antioxidant markers such as malondialdhyde (MDA) and superoxide dismutase (SOD) and fibrotic markers such as hyaluronic acid (HA) and procollagen-III (procol-III) were evaluated in liver tissues. IL-10 as well as PGE2 were also assayed in liver tissues. Pathologic changes in the liver were detected by hematoxylin and eosin staining. Collagen precipitation in liver tissues was visualized using masson trichrom stain. Results: Rebamipide inhibit CCL4 induced increase in ALT and AST significantly (p < 0.05). Rebamipide exerted an antioxidant effect as it inhibits CCL4 induced increased MDA level and decreased SOD activity. Fibrotic markers assay revealed that repamipide (60 or 100 mg/kg/day) decreased the level of procol-III and HA compared to CCl4 (p < 0.05). Oral administration of Rebamipide was associated with a significant increase (p < 0.05) of PGE2 and IL-10. Rebamipide especially at the dose of (100 mg/kg/day) restores liver histology structure and abolish collagen precipitation in liver tissues. Conclusion: Rebamipide retards hepatic fibrosis induced by CCL4 may be through the induction of PGE2 level.Keywords: fibrotic markers, hepatic fibrosis, PGE2, rebamipide
Procedia PDF Downloads 484213 DUSP16 Inhibition Rescues Neurogenic and Cognitive Deficits in Alzheimer's Disease Mice Models
Authors: Huimin Zhao, Xiaoquan Liu, Haochen Liu
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The major challenge facing Alzheimer's Disease (AD) drug development is how to effectively improve cognitive function in clinical practice. Growing evidence indicates that stimulating hippocampal neurogenesis is a strategy for restoring cognition in animal models of AD. The mitogen-activated protein kinase (MAPK) pathway is a crucial factor in neurogenesis, which is negatively regulated by Dual-specificity phosphatase 16 (DUSP16). Transcriptome analysis of post-mortem brain tissue revealed up-regulation of DUSP16 expression in AD patients. Additionally, DUSP16 was involved in regulating the proliferation and neural differentiation of neural progenitor cells (NPCs). Nevertheless, whether the effect of DUSP16 on ameliorating cognitive disorders by influencing NPCs differentiation in AD mice remains unclear. Our study demonstrates an association between DUSP16 SNPs and clinical progression in individuals with mild cognitive impairment (MCI). Besides, we found that increased DUSP16 expression in both 3×Tg and SAMP8 models of AD led to NPC differentiation impairments. By silencing DUSP16, cognitive benefits, the induction of AHN and synaptic plasticity, were observed in AD mice. Furthermore, we found that DUSP16 is involved in the process of NPC differentiation by regulating c-Jun N-terminal kinase (JNK) phosphorylation. Moreover, the increased DUSP16 may be regulated by the ETS transcription factor (ELK1), which binds to the promoter region of DUSP16. Loss of ELK1 resulted in decreased DUSP16 mRNA and protein levels. Our data uncover a potential regulatory role for DUSP16 in adult hippocampal neurogenesis and provide a possibility to find the target of AD intervention.Keywords: alzheimer's disease, cognitive function, DUSP16, hippocampal neurogenesis
Procedia PDF Downloads 72212 Potential Therapeutic Effect of Obestatin in Oral Mucositis
Authors: Agnieszka Stempniewicz, Piotr Ceranowicz, Wojciech Macyk, Jakub Cieszkowski, Beata Kuśnierz-Cabała, Katarzyna Gałązka, Zygmunt Warzecha
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Objectives: There are numerous strategies for the prevention or treatment of oral mucositis. However, their effectiveness is limited and does not correspond to expectations. Recent studies have shown that obestatin exhibits a protective effect and accelerates the healing of gastrointestinal mucosa. The aim of the present study was to examine the influence of obestatin administration on oral ulcers in rats. Methods: lingual ulcers were induced by the use of acetic acid. Rats were treated twice a day intraperitoneally with saline or obestatin(4, 8, or 16 nmol/kg/dose) for five days. The study determined: lingual mucosa morphology, cell proliferation, mucosal blood flow, and mucosal pro-inflammatory interleukin-1β level(IL-1β). Results: In animals without induction of oral ulcers, treatment with obestatin was without any effect. Obestatin administration in rats with lingual ulcers increased the healing rate of these ulcers. Obestatin given at the dose of 8 or 16 nmol/kg/dose caused the strongest and similar therapeutic effect. This result was associated with a significant increase in blood flow and cell proliferation in gingival mucosa, as well as a significant decrease in IL-1β level. Conclusions: Obestatin accelerates the healing of lingual ulcers in rats. This therapeutic effect is well-correlated with an increase in blood flow and cell proliferation in oral mucosa, as well as a decrease in pro-inflammatory IL-1β levels. Obestatin is a potentially useful candidate for the prevention and treatment of oral mucositis. Acknowledgment: Agnieszka Stempniewicz acknowledges the support of InterDokMed project no. POWR.03.02.00- 00-I013/16.Keywords: oral mucositis, ulcers, obestatin, lingual mucosa
Procedia PDF Downloads 73211 The Effect of Jujube Extract and Resistance Training on the Reduction of Complications Caused by the Induction of Anabolic Steroid Boldenone on the Histopathological Changes of Pancreatic Tissue of Male Wistar Rats
Authors: Sayyed-javad Ziaolhagh, Ali-Reza Saadatifar
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Introduction: Athletes frequently perform anabolic steroid resistance exercise, but the effects of medical doses and abuse along with resistance exercise on structural damage to the Pancreases and also jujube extract are unknown. The aim of this study was to investigate the effects of resistance training on body weight and hip fractures induced by boldenone injection in male rats. Materials and methods: In this experimental study, 30 male Wistar rats aged 8-12 weeks (weight 202±9.34 g) were randomly divided into five groups: control, boldenone, extract of iujuba+boldenone, boldenone+resistance training and boldenone+resistance training +extract of jujuba. The resistance training program included climbing the ladder for 8 weeks, 3 days a week, 1 session training in a day and each session consisted of the 3 sets and 5 repetitions. Injection was conducted in depth in the hamstring once a week on an appointed day. After anesthesia, autopsy was performed, and the cardiac tissue was isolated. Results: Results showed that boldenone caused tissue damage, congestion, and nuclei unclear and diffuse. In the group "resistance + Boldenone," The Pancreases tissue showed a high degree of hyperemia, and the muscle cells were somewhat abnormal. In boldenone + jujube, the appearance of the tissue was normal, and the rejuvenating effect was visible. Conclusion: Boldenone appears to cause structural damage to the Pancreases tissue. Strength training with Jujube Extract can reduce part of the pancreatic system disorders (necrosis and inflammation) caused by anabolic steroid use.Keywords: boldenone, Jujube extract, pancreases tissue, resistance training
Procedia PDF Downloads 70210 Linking Metabolism, Pluripotency and Epigenetic Changes during Early Differentiation of Embryonic Stem Cells
Authors: Arieh Moussaieff, Bénédicte Elena-Herrmann, Yaakov Nahmias, Daniel Aberdam
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Differentiation of pluripotent stem cells is a slow process, marked by the gradual loss of pluripotency factors over days in culture. While the first few days of differentiation show minor changes in the cellular transcriptome, intracellular signaling pathways remain largely unknown. Recently, several groups demonstrated that the metabolism of pluripotent mouse and human cells is different from that of somatic cells, showing a marked increase in glycolysis previously identified in cancer as the Warburg effect. Here, we sought to identify the earliest metabolic changes induced at the first hours of differentiation. High-resolution NMR analysis identified 35 metabolites and a distinct, gradual transition in metabolism during early differentiation. Metabolic and transcriptional analyses showed the induction of glycolysis toward acetate and acetyl-coA in pluripotent cells, and an increase in cholesterol biosynthesis during early differentiation. Importantly, this metabolic pathway regulated differentiation of human and mouse embryonic stem cells. Acetate delayed differentiation preventing differentiation-induced histone de-acetylation in a dose-dependent manner. Glycolytic inhibitors upstream of acetate caused differentiation of pluripotent cells, while those downstream delayed differentiation. Our data suggests that a rapid loss of glycolysis in early differentiation down-regulates acetate and acetyl-coA production, causing a loss of histone acetylation and concomitant loss of pluripotency. It demonstrate that pluripotent stem cells utilize a novel metabolism pathway to maintain pluripotency through acetate/acetyl-coA and highlights the important role metabolism plays in pluripotency and early differentiation of stem cells.Keywords: pluripotency, metabolomics, epigenetics, acetyl-coA
Procedia PDF Downloads 470209 Evaluation of Chemopreventive Activity of Medicinal Plant, Gromwell Seed against Tumor Promoting Stage
Authors: Harukuni Tokuda, Takanari Arai, Xu FengHao, Nobutaka Suzuki
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In our continuous search for anti-tumor promoting, chemopreventive active potency from natural source material, a kind of healthy tea, Gromwell seed (Coix lachryma-jobi) ext., and including compounds Monoolein and Trilinolein have been screened using the in vitro synergistic assay indicated by inhibitory effects on the induction of Epstein-Barr virus early antigen (EBV-EA) by TPA. In assay, Gromwell seed aqueous extract and hot aqueous extract exhibited the potential inhibitory effects on EBV-EA activation without strong cytotoxicity on Raji cells. In our experimental system, the inhibitory effects of both Gromwell extracts and compounds were greater than that of beta-carotene, which is known anti-tumor promoting agent and/or chemopreventive agent. These compounds were evaluated for their in vitro inhibitory effect on EBV-EA activation induced by TPA. The percentages of the inhibition of TPA-induced EBV-EA activation for these materials were 60% and 30% at concentration 100 μg. Based on the results obtained in vitro, we studied the inhibitory effect of compounds, in an in vivo two-stage carcinogenesis test of mouse skin papilloma using DMBA as an initiator and TPA as a potential promoter. The control animals showed a 100% incidence of papilloma at 20 weeks after DMBA-TPA tumor promotion, while treatment with compounds reduced the percentage of number of tumor to 60 % after 20 weeks. Results from in vitro and in vivo studies showing chemopreventive activity against TPA promoting stage and these data support the effective potency of carcinogenic stage in clinical evaluation of integrative oncology.Keywords: gromwell seed, medicinal plant, chemoprevention, pharmaceutical medicine
Procedia PDF Downloads 406208 Effect of High Intensity Ultrasonic Treatment on the Micro Structure, Corrosion and Mechanical Behavior of ac4c Aluminium Alloy
Authors: A.Farrag Farrag, A. M. El-Aziz Abdel Aziz, W. Khlifa Khlifa
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Ultrasonic treatment is a promising process nowadays in the engineering field due to its high efficiency and it is a low-cost process. It enhances mechanical properties, corrosion resistance, and homogeneity of the microstructure. In this study, the effect of ultrasonic treatment and several casting conditions on microstructure, hardness and corrosion behavior of AC4C aluminum alloy was examined. Various ultrasonic treatments of the AC4C alloys were carried out to prepare billets for thixocasting process. Treatment temperatures varied from about 630oC and cooled down to under ultrasonic field. Treatment time was about 90s. A 600-watts ultrasonic system with 19.5 kHz and intensity of 170 W/cm2 was used. Billets were reheated to semisolid state and held for 5 minutes at 582 oC and temperatures (soaking) using high-frequency induction system, then thixocasted using a die casting machine. Microstructures of the thixocast parts were studied using optical and SEM microscopes. On the other hand, two samples were conventionally cast and poured at 634 oC and 750 oC. The microstructure showed a globular none dendritic grains for AC4C with the application of UST at 630-582 oC, Less dendritic grains when the sample was conventionally cast without the application of UST and poured at 624 oC and a fully dendritic microstructure When the sample was cast and poured at 750 oC without UST .The ultrasonic treatment during solidification proved that it has a positive influence on the microstructure as it produced the finest and globular grains thus it is expected to increase the mechanical properties of the alloy. Higher values of corrosion resistance and hardness were recorded for the ultrasound-treated sample in comparison to cast one.Keywords: ultrasonic treatment, aluminum alloys, corrosion behaviour, mechanical behaviour, microstructure
Procedia PDF Downloads 353207 Mediation Role of Teachers’ Surface Acting and Deep Acting on the Relationship between Calling Orientation and Work Engagement
Authors: Yohannes Bisa Biramo
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This study examined the meditational role of surface acting and deep acting on the relationship between calling orientation and work engagement of teachers in secondary schools of Wolaita Zone, Wolaita, Ethiopia. A predictive non-experimental correlational design was performed among 300 secondary school teachers. Stratified random sampling followed by a systematic random sampling technique was used as the basis for selecting samples from the target population. To analyze the data, Structural Equation Modeling (SEM) was used to test the association between the independent variables and the dependent variables. Furthermore, the goodness of fit of the study variables was tested using SEM to see and explain the path influence of the independent variable on the dependent variable. Confirmatory factor analysis (CFA) was conducted to test the validity of the scales in the study and to assess the measurement model fit indices. The analysis result revealed that calling was significantly and positively correlated with surface acting, deep acting and work engagement. Similarly, surface acting was significantly and positively correlated with deep acting and work engagement. And also, deep acting was significantly and positively correlated with work engagement. With respect to mediation analysis, the result revealed that surface acting mediated the relationship between calling and work engagement and also deep acting mediated the relationship between calling and work engagement. Besides, by using the model of the present study, the school leaders and practitioners can identify a core area to be considered in recruiting and letting teachers teach, in giving induction training for newly employed teachers and in performance appraisal.Keywords: calling, surface acting, deep acting, work engagement, mediation, teachers
Procedia PDF Downloads 81206 CP-96345 Rregulates Hydrogen Sulphide Induced TLR4 Signaling Pathway Adhesion Molecules in Caerulein Treated Pancreatic Acinar Cells
Authors: Ramasamy Tamizhselvi, Leema George, Madhav Bhatia
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We have earlier shown that mouse pancreatic acinar cells produce hydrogen sulfide (H2S) and play a role in the pathogenesis of acute pancreatitis. This study is to determine the effect of H2S on TLR4 mediated innate immune signaling in acute pancreatitis via substance P (SP). Male Swiss mice were treated with hourly intraperitoneal injection of caerulein (50μg/kg) for 10 hour. DL-propargylglycine (PAG) (100 mg/kg i.p.), an inhibitor of H2S formation was administered 1h after the induction of acute pancreatitis. Pancreatic acinar cells from male Swiss mice were incubated with or without caerulein (10–7 M for 60 min) and CP-96345 (NK1R inhibitor). To better understand the effect of H2S in inflammation, acinar cells were stimulated with caerulein after addition of H2S donor, NaHS. In addition, caerulein treated pancreatic acinar cells were pretreated with PAG (30 µM), for 1h. H2S inhibitor, PAG, eliminated TLR4, IRAK4, TRAF6 and NF-kB levels in an in vitro and in vivo model of caerulein-induced acute pancreatitis. PPTA gene deletion reduced TLR4, MyD88, IRAK4, TRAF6, adhesion molecules and NF-kB in caerulein treated pancreatic acinar cells whereas administration of NaHS resulted in further rise in TLR4 and NF-kB levels in caerulein treated pancreatic acinar cells. In addition, acini isolated from mice and treated with PPTA gene receptor NK1R antagonist CP96345 did not exhibit further increase in TLR4, IRAK4, TRAF6, adhesion molecules and NF-kB levels after NaHS pretreatment. The present findings show for the first time that in acute pancreatitis, H2S up-regulates TLR4 pathway and NF-kB via substance P.Keywords: preprotachykinin-A gene, H2S, TLR4, acute pancreatitis
Procedia PDF Downloads 276205 Albendazole Ameliorates Inflammatory Response in a Rat Model of Acute Mesenteric Ischemia Reperfusion Injury
Authors: Kamyar Moradi
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Background: Acute mesenteric ischemia is known as a life-threatening condition. Re-establishment of blood flow in this condition can lead to mesenteric ischemia reperfusion (MIR) injury, which is accompanied by inflammatory response. Still, clear blueprint of inflammatory mechanism underlying MIR injury has not been provided. Interestingly, Albendazole has exhibited notable effects on inflammation and cytokine production. In this study, we aimed to evaluate outcomes of MIR injury following pretreatment with Albendazole with respect to assessment of mesenteric inflammation and ischemia threshold. Methods: Male rats were randomly divided into sham operated, vehicle treated, Albendazole 100 mg/kg, and Albendazole 200 mg/kg groups. MIR injury was induced by occlusion of superior mesenteric artery for 30 minutes followed by 120 minutes of reperfusion. Samples were utilized for assessment of epithelial survival and villous height. Immunohistochemistry study revealed intestinal expression of TNF-α and HIF-1-α. Gene expression of NF-κB/TLR4/TNF-α/IL-6 was measured using RTPCR. Also, protein levels of inflammatory cytokines in serum and intestine were assessed by ELISA method. Results: Histopathological study demonstrated that pretreatment with Albendazole could ameliorate decline in villous height and epithelial survival following MIR injury. Also, systemic inflammation was suppressed after administration of Albendazole. Analysis of possible participating inflammatory pathway could demonstrate that intestinal expression of NF-κB/TLR4/TNF-α/IL-6 is significantly attenuated in treated groups. Eventually, IHC study illustrated concordant decline in mesenteric expression of HIF-1-α/TNF-α. Conclusion: Single dose pretreatment with Albendazole could ameliorate inflammatory response and enhance ischemia threshold following induction of MIR injury. Still, more studies would clarify existing causality in this phenomenon.Keywords: albendazole, ischemia reperfusion injury, inflammation, mesenteric ischemia
Procedia PDF Downloads 169204 Induction of G1 Arrest and Apoptosis in Human Cancer Cells by Panaxydol
Authors: Dong-Gyu Leem, Ji-Sun Shin, Sang Yoon Choi, Kyung-Tae Lee
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In this study, we focused on the anti-proliferative effects of panaxydol, a C17 polyacetylenic compound derived from Panax ginseng roots, against various human cancer cells. We treated with panaxydol to various cancer cells and panaxydol treatment was found to significantly inhibit the proliferation of human lung cancer cells (A549) and human pancreatic cancer cells (AsPC-1 and MIA PaCa-2), of which AsPC-1 cells were most sensitive to its treatment. DNA flow cytometric analysis indicated that panaxydol blocked cell cycle progression at the G1 phase in A549 cells, which accompanied by a parallel reduction of protein expression of cyclin-dependent kinase (CDK) 2, CDK4, CDK6, cyclin D1 and cyclin E. CDK inhibitors (CDKIs), such as p21CIP1/WAF1 and p27KIP1, were gradually upregulated after panaxydol treatment at the protein levels. Furthermore, panaxydol induced the activation of p53 in A549 cells. In addition, panaxydol also induced apoptosis of AsPC-1 and MIA PaCa-2 cells, as shown by accumulation of subG1 and apoptotic cell populations. Panaxydol triggered the activation of caspase-3, -8, -9 and the cleavage of poly (ADP-ribose) polymerase (PARP). Reduction of mitochondrial transmembrane potential by panaxydol was determined by staining with dihexyloxacarbocyanine iodide. Furthermore, panaxydol suppressed the levels of anti-apoptotic proteins, XIAP and Bcl-2, and increased the levels of proapoptotic proteins, Bax and Bad. In addition, panaxydol inhibited the activation of Akt and extracellular signal-regulated kinase (ERK) and activated the p38 mitogen-activated protein kinase kinase (MAPK). Our results suggest that panaxydol is an anti-tumor compound that causes p53-mediated cell cycle arrest and apoptosis via mitochondrial apoptotic pathway in various cancer cells.Keywords: apoptosis, cancer, G1 arrest, panaxydol
Procedia PDF Downloads 322203 Investigation of Free Vibrations of Opened Shells from Alloy D19: Assistance of the Associated Mass System
Authors: Oleg Ye Sysoyev, Artem Yu Dobryshkin, Nyein Sitt Naing
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Cylindrical shells are widely used in the construction of buildings and structures, as well as in the air structure. Thin-walled casings made of aluminum alloys are an effective substitute for reinforced concrete and steel structures in construction. The correspondence of theoretical calculations and the actual behavior of aluminum alloy structures is to ensure their trouble-free operation. In the laboratory of our university, "Building Constructions" conducted an experimental study to determine the effect of the system of attached masses on the natural oscillations of shallow cylindrical shells of aluminum alloys, the results of which were compared with theoretical calculations. The purpose of the experiment is to measure the free oscillations of an open, sloping cylindrical shell for various variations of the attached masses. Oscillations of an open, slender, thin-walled cylindrical shell, rectangular in plan, were measured using induction accelerometers. The theoretical calculation of the shell was carried out on the basis of the equations of motion of the theory of shallow shells, using the Bubnov-Galerkin method. A significant splitting of the flexural frequency spectrum is found, influenced not only by the systems of attached маsses but also by the values of the wave formation parameters, which depend on the relative geometric dimensions of the shell. The correspondence of analytical and experimental data is found, using the example of an open shell of alloy D19, which allows us to speak about the high quality of the study. A qualitative new analytical solution of the problem of determining the value of the oscillation frequency of the shell, carrying a system of attached masses is shown.Keywords: open hollow shell, nonlinear oscillations, associated mass, frequency
Procedia PDF Downloads 295202 Evaluation of the Ability of COVID-19 Infected Sera to Induce Netosis Using an Ex-Vivo NETosis Monitoring Tool
Authors: Constant Gillot, Pauline Michaux, Julien Favresse, Jean-Michel Dogné, Jonathan Douxfils
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Introduction: NETosis has emerged as a crucial yet paradoxical factor in severe COVID-19 cases. While neutrophil extracellular traps (NETs) help contain and eliminate viral particles, excessive NET formation can lead to hyperinflammation, exacerbating tissue damage and acute respiratory distress syndrome (ARDS). Aims: This study evaluates the relationship between COVID-19-infected sera and NETosis using an ex-vivo model. Methods: Sera from 8 post-admission COVID-19 patients, after receiving corticoid therapy, were used to induce NETosis in neutrophils from a healthy donor. NET formation was tracked using fluorescent markers for DNA and neutrophil elastase (NE) every 2 minutes for 8 hours. The results were expressed as a percentage of DNA/NE released over time. Key metrics, including T50 (time to 50% release) and AUC (area under the curve), representing total NETosis potential), were calculated. A 27-cytokine screening kit was used to assess the cytokine composition of the sera. Results: COVID-19 sera induced NETosis based on their cytokine profile. The AUC of NE and DNA release decreased with time following corticoid therapy, showing a significant reduction in 6 of the 8 patients (p<0.05). T50 also decreased in parallel with AUC for both markers. Cytokines concentration decrease with time after therapy administration. There is correlation between 14 cytokines concentration and NE release. Conclusion: This ex-vivo model successfully demonstrated the induction of NETosis by COVID-19 sera using two markers. A clear decrease in NETosis potential was observed over time with glucocorticoid therapy. This model can be a valuable tool for monitoring NETosis and investigating potential NETosis inducers and inhibitors.Keywords: NETosis, COVID-19, cytokine storm, biomarkers
Procedia PDF Downloads 19201 Deubiquitinase USP35 Regulates Mitosis Progression by Blocking CDH1-Mediated Degradation of Aurora B.
Authors: Jinyoung Park, Eun Joo Song
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Introduction: Deubiquitinating enzymes (DUBs) are proteases that cleave ubiquitin or ubiquitin-like modifications on substrates. Deubiquitination could regulate cellular physiology, such as signal transduction, DNA damage and repair, and cell cycle progression. Although more than 100 DUBs are encoded in the human and the importance of DUBs has been realized, the functions of most DUBs are unknown. This study aims to identify the molecular mechanism by which deubiquitinating enzyme USP35 regulates cell cycle progression for the first time. Methods: USP35 RNAi was mainly used to identify the function of USP35 in cell cycle progression. To find substrates of USP35, we analyzed protein-protein interaction using LC-MS. Several biological methods, such as ubiquitination assay, cell synchronization, immunofluorescence, and immunoprecipitation assay were used to investigate the exact mechanism by which USP35 affects successful completion of mitosis. Results: USP35 knockdown caused not only reduction of mitotic cell number but also induction of mitotic cells with abnormal spindle formation. Actually, cell proliferation was decreased by USP35 knockdown. Interestingly, we found that loss of USP35 decreased the stability and expression of Aurora B, a member of chromosomal passenger complex (CPC), and the phosphorylation of its substrate. Indeed, USP35 interacted with Aurora B and deubiquitinated it. In addition, USP35 knockdown induced abnormal localization of Aurora B in mitotic cells. Finally, CDH1-mediated ubiquitination of Aurora B level was rescued by USP35 overexpression, but not inactive form of USP35, USP35 C450A. Discussion: Our findings suggest that USP35 regulates Aurora B-mediated mitotic spindle assembly and G2-M transition by blocking CDH1-induced degradation of Aurora B.Keywords: USP35, HSP90, Aurora B, cell cycle progression
Procedia PDF Downloads 358200 Involvement of BCRP/ABCG2 in Protective Mechanisms of Resveratrol against Methotrexate-Induced Renal Damage in Rats
Authors: Mohamed A. Morsy, Azza A. El-Sheikh, Abdulla Y. Al-Taher
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Resveratrol (RES) is a well-known polyphenol antioxidant. We have previously shown that testicular protective effect of RES against the anticancer drug methotrexate (MTX)-induced toxicity involves transporter-mediated mechanisms. Here, we investigated the effect of RES on MTX-induced nephrotoxicity. Rats were administered RES (10 mg/kg/day) for 8 days, with or without a single MTX dose (20 mg/kg i.p.) at day 4 of the experiment. MTX induced nephrotoxicity evident by significantly increase in serum blood urea nitrogen and creatinine compared to control, as well as distortion of kidney microscopic structure. MTX also significantly increased renal nitric oxide level, with induction of inducible nitric oxide synthase expression. MTX also significantly up-regulated fas ligand and caspase 3. Administering RES prior to MTX significantly improved kidney function and microscopic picture, as well as significantly decreased nitrosative and apoptotic markers compared to MTX alone. RES, but not MTX, caused significant increase in expression of breast cancer resistance protein (BCRP), an apical efflux renal transporter that participates in urinary elimination of both MTX and RES. Interestingly, concomitant MTX and RES caused further up-regulation of renal Bcrp compared to RES alone. Using Human BCRP ATPase assay, both RES and MTX exhibited dose-dependent increase in ATPase activity, with Km values of 0.52 ± 0.03 and 30.9 ± 4.2 µM, respectively. Furthermore, combined RES and MTX caused ATPase activity which was significantly less than maximum ATPase activity attained by the positive control; sulfasalazine (12.5 µM). In conclusion, RES exerted nephro-protection against MTX-induced toxicity through anti-nitrosative and anti-apoptotic effects, as well as via up-regulation of renal Bcrp.Keywords: methotrexate, resveratrol, nephrotoxicity, breast cancer resistance protein
Procedia PDF Downloads 295199 Hyparrhenia hirta: A Potential Protective Agent against DNA Damage and Liver Toxicity of Sodium Nitrate in Adult Rats
Authors: Hanen Bouaziz-Ketata, Ghada Ben Salah, Hichem Ben Salah, Kamel Jamoussi, Najiba Zeghal
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The present study investigated the protective role of Hyparrhenia hirta on nitrate-induced liver damage. Experiments were carried out on adult rats divided into 3 groups, a control group and two treated groups. NaNO3 was administered daily by oral gavage at a dose of 400 mg/kg bw in treated groups either alone or coadministered with Hyparrhenia hirta methanolic extract via drinking water at a dose of 200 mg/kg bw for 50 days. Liver toxicity induced by NaNO3 was characterized by higher serum levels of glucose, total cholesterol and triglyceride and lower serum total protein than those of controls. Transaminases and lactate deshydrogenase activities in serum were elevated indicating hepatic cells’ damage after treatment with NaNO3. The hyperbilirubinemia and the increased serum gamma glutamyl transferase activities suggested the presence of cholestasis in NaNO3 exposed rats. In parallel, NaNO3 caused oxidant/antioxidant imbalance in the liver as reflected by the increased lipid peroxidation, the decreased total glutathione content and superoxide dismutase, catalase and glutathione peroxidase activities. Nitrate caused also a significant induction of DNA fragmentation as evidenced by the presence of a smear without ladder formation on agarose gel. Hyparrhenia hirta supplementation showed an improvement of all parameters cited above. We conclude that the present work provides ethnopharmacological relevance of Hyparrhenia hirta against the toxic effect of nitrate, suggesting its role as a potential antioxidant.Keywords: Hyparrhenia hirta, liver, nitrate toxicity, oxidative stress, rat
Procedia PDF Downloads 545198 Molecular Modeling a Tool for Postulating the Mechanism of Drug Interaction: Glimepiride Alters the Pharmacokinetics of Sildenafil Citrate in Diabetic Nephropathy Animals
Authors: Alok Shiomurti Tripathi, Ajay Kumar Timiri, Papiya Mitra Mazumder, Anil Chandewar
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The present study evaluates the possible drug interaction between glimepiride (GLIM) and sildenafil citrate (SIL) in streptozotocin (STZ) induced in diabetic nephropathic (DN) animals and also postulates the possible mechanism of interaction by molecular modeling studies. Diabetic nephropathy was induced by single dose of STZ (60 mg/kg, ip) and confirms it by assessing the blood and urine biochemical parameters on 28th day of its induction. Selected DN animals were used for the drug interaction between GLIM (0.5mg/kg, p.o.) and SIL (2.5 mg/kg, p.o.) after 29th and 70th day of protocol. Drug interaction were assessed by evaluating the plasma drug concentration using HPLC-UV and also determine the change in the biochemical parameter in blood and urine. Mechanism of the interaction was postulated by molecular modeling study using Maestro module of Schrodinger software. DN was confirmed as there was significant alteration in the blood and urine biochemical parameter in STZ treated groups. The concentration of SIL increased significantly (p<0.001) in rat plasma when co administered with GLIM after 70th day of protocol. Molecular modelling study revealed few important interactions with rat serum albumin and CYP2C9.GLIM has strong hydrophobic interaction with binding site residues of rat serum albumin compared to SIL. Whereas, for CYP2C9, GLIM has strong hydrogen bond with polar contacts and hydrophobic interactions than SIL. Present study concludes that bioavailability of SIL increases when co-administered chronically with GLIM in the management of DN animals and mechanism has been supported by molecular modeling studies.Keywords: diabetic nephropathy, glimepiride, sildenafil citrate, pharmacokinetics, homology modeling, schrodinger
Procedia PDF Downloads 378197 Analysis of the Influence of Fiber Volume and Fiber Orientation on Post-Cracking Behavior of Steel Fiber Reinforced Concrete
Authors: Marilia M. Camargo, Luisa A. Gachet-Barbosa, Rosa C. C. Lintz
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The addition of fibers into concrete matrix can enhance some properties of the composite, such as tensile, flexural and impact strengths, toughness, deformation capacity and post-cracking ductility. Many factors affect the mechanical behavior of fiber reinforced concrete, such as concrete matrix (concrete strength, additions, aggregate diameter, etc.), characteristics of the fiber (geometry, type, aspect ratio, volume, orientation, distribution, strength, stiffness, etc.), specimen (size, geometry, method of preparation and loading rate). This research investigates the effects of fiber volume and orientation on the post-cracking behavior of steel fiber reinforced concrete (SFRC). Hooked-end steel fibers with aspect ratios of 45 were added into concrete with volume of 0,32%, 0,64%, 0,94%. The post-cracking behaviour was assessed by double punch test of cubic specimens and the actual volume and orientation of the fibers were determined by non-destructive tests by means of electromagnetic induction. The results showed that the actual volume of fibers in each sample differs in a small amount from the dosed volume of fibers and that the deformation and toughness of the concrete increase with the increase in the actual volume of fibers. In determining the orientation of the fibers, it was found that they tend to distribute more in the X and Y axes due to the influence of the walls of the mold. In addition, it was concluded that the orientation of the fibers is important in the post-cracking behaviour of FRC when analyzed together with the actual volume of fibers, since the greater the volume of fibers, the greater the number of fibers oriented orthogonally to the application of loadings and, consequently, there is a better mechanical behavior of the composite. These results provide a better understanding of the influence of volume and fiber orientation on the post-cracking behavior of the FRC.Keywords: fiber reinforced concrete, steel fibers, volume of fibers, orientation of fibers, post-cracking behaviour
Procedia PDF Downloads 179196 Treatment of Feline Infectious Peritonitis in Cats with Molnupiravir: Clinical Observations and Outcomes For 54 Cases
Authors: T. M. Clark, S. J. Coggins, R. Malik, J. King, R. Korman
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Objectives: This observational study investigated the use of molnupiravir for treatment of cats with naturally occurring feline infectious peritonitis. Methods: From September 2022 to February 2024, 66 cats diagnosed with FIP across 32 veterinary practices, mainly in Australia, were enrolled. Of these, 54 cats met the inclusion criteria. Complete remission was defined by the resolution of clinical signs and normalisation of A:G ratio (to ≥0.6). Presumptive remission was defined as sustained resolution of FIP-related clinical signs for at least 100 days post cessation of antiviral therapy. Results: In Cohort 1, 18 cats were treated with molnupiravir monotherapy. Thirteen achieved complete remission and three attained presumptive remission, resulting in an overall remission rate of 89% and a provisional cure rate of 72%, with three relapses. Cohort 2 included 29 cats treated with a short induction course of GS-441524 and/or remdesivir before switching to molnupiravir; 23 attained complete remission, and two achieved presumptive remission. The overall cure rate was 86% with no relapses. Seven cats in cohort 3 were initially treated with extended courses of GS-441524, remdesivir, and/or mefloquine and experienced treatment failure or relapse. Molnupiravir was introduced as a rescue therapy; 6 achieved complete remission and 1 achieved presumed remission, resulting in a 100% cure rate with no relapses. Few adverse effects were reported, with the most notable including neutropenia, transient elevations in hepatic enzymes, and polydipsia/polyuria. Conclusion and Relevance: Molnupiravir as a monotherapy, or in combination with other antivirals, represents an accessible, effective treatment for FIP when given at a dosage of 10-15 mg/kg BID. Success occurred across various presentations of FIP, including cases with ocular and neurological involvement.Keywords: feline infectious peritonitis, FIP, molnupiravir, nucleoside analogue, antiviral
Procedia PDF Downloads 18195 A Structure-Switching Electrochemical Aptasensor for Rapid, Reagentless and Single-Step, Nanomolar Detection of C-Reactive Protein
Authors: William L. Whitehouse, Louisa H. Y. Lo, Andrew B. Kinghorn, Simon C. C. Shiu, Julian. A. Tanner
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C-reactive protein (CRP) is an acute-phase reactant and sensitive indicator for sepsis and other life-threatening pathologies, including systemic inflammatory response syndrome (SIRS). Currently, clinical turn-around times for established CRP detection methods take between 30 minutes to hours or even days from centralized laboratories. Here, we report the development of an electrochemical biosensor using redox probe-tagged DNA aptamers functionalized onto cheap, commercially available screen-printed electrodes. Binding-induced conformational switching of the CRP-targeting aptamer induces a specific and selective signal-ON event, which enables single-step and reagentless detection of CRP in as little as 1 minute. The aptasensor dynamic range spans 5-1000nM (R=0.97) or 5-500nM (R=0.99) in 50% diluted human serum, with a LOD of 3nM, corresponding to 2-orders of magnitude sensitivity under the clinically relevant cut-off for CRP. The sensor is stable for up to one week and can be reused numerous times, as judged from repeated real-time dosing and dose-response assays. By decoupling binding events from the signal induction mechanism, structure-switching electrochemical aptamer-based sensors (SS-EABs) provide considerable advantages over their adsorption-based counterparts. Our work expands on the retinue of such sensors reported in the literature and is the first instance of an SS-EAB for reagentless CRP detection. We hope this study can inspire further investigations into the suitability of SS-EABs for diagnostics, which will aid translational R&D toward fully realized devices aimed at point-of-care applications or for use more broadly by the public.Keywords: structure-switching, C-reactive protein, electrochemical, biosensor, aptasensor.
Procedia PDF Downloads 70194 Phytochemical Screening, Antioxidant and Hepatoprotection Assessment of Extracts of Coriandrum sativm L. on Wistar Rats
Authors: Hiba T. Allah ALtieb Gusm ALsied, Amna Beshir Medani Ahmed, Ikram Mohamed ELtayeb, Saad Mohamed Hussein Ayoub
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This study was carried out to determine the hepatoactivity and the antioxidant activity of Coiradrum sativum L. aerial part and fruit extracts against CCL4 induced acute liver damages in Wistar rats. The aerial parts and fruits part of the plant were extracted 96% ethanol with soxhlet apparatus. Hepatic injury was achieved by subcutaneous injection of 3 ml/kg of CCL4 diluted with olive oil with ratio 1:1. The extracts were mixed together 1:1 ratio and given in different doses 100,200,400 mg/kg/day for 5 days under CCL4 induction at 3rd day. The significance of differences between means by using T-test was compared among the groups. The mixture of the two extracts didn’t show any significant result in protecting liver injury (antagonistic effects), it shows high level of liver enzyme like alkaline phosphatase (ALP), glutamate oxaloacetate transaminase (SGOT) and glutamate pyruvate transaminase (SGPT). Serological studies further confirmed the results. The results obtained were compared with silymarin (70 mg/kg/day) orally, the standard drug for hepatoprotection which show recovery close to normalization almost like that of silymarin; therefore, further studies on this plant with different ratios especially in isolated tissue to spot more light on antagonistic effects of the two extracts. Antioxidant activity of the extracts was determined by the DPPH method. The results obtained show high anti-oxidant activity for fruits extract while slight or moderate antioxidant activity to aerial extracts.Keywords: antioxidant, aerial part, Coriadrum sativum L., fruity, hepatoprotection, Silymarin, phytochemical screening
Procedia PDF Downloads 489193 Design Components and Reliability Aspects of Municipal Waste Water and SEIG Based Micro Hydro Power Plant
Authors: R. K. Saket
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This paper presents design aspects and probabilistic approach for generation reliability evaluation of an alternative resource: municipal waste water based micro hydro power generation system. Annual and daily flow duration curves have been obtained for design, installation, development, scientific analysis and reliability evaluation of the MHPP. The hydro potential of the waste water flowing through sewage system of the BHU campus has been determined to produce annual flow duration and daily flow duration curves by ordering the recorded water flows from maximum to minimum values. Design pressure, the roughness of the pipe’s interior surface, method of joining, weight, ease of installation, accessibility to the sewage system, design life, maintenance, weather conditions, availability of material, related cost and likelihood of structural damage have been considered for design of a particular penstock for reliable operation of the MHPP. A MHPGS based on MWW and SEIG is designed, developed, and practically implemented to provide reliable electric energy to suitable load in the campus of the Banaras Hindu University, Varanasi, (UP), India. Generation reliability evaluation of the developed MHPP using Gaussian distribution approach, safety factor concept, peak load consideration and Simpson 1/3rd rule has presented in this paper.Keywords: self excited induction generator, annual and daily flow duration curve, sewage system, municipal waste water, reliability evaluation, Gaussian distribution, Simpson 1/3rd rule
Procedia PDF Downloads 558192 Biophysical Study of the Interaction of Harmalol with Nucleic Acids of Different Motifs: Spectroscopic and Calorimetric Approaches
Authors: Kakali Bhadra
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Binding of small molecules to DNA and recently to RNA, continues to attract considerable attention for developing effective therapeutic agents for control of gene expression. This work focuses towards understanding interaction of harmalol, a dihydro beta-carboline alkaloid, with different nucleic acid motifs viz. double stranded CT DNA, single stranded A-form poly(A), double-stranded A-form of poly(C)·poly(G) and clover leaf tRNAphe by different spectroscopic, calorimetric and molecular modeling techniques. Results of this study converge to suggest that (i) binding constant varied in the order of CT DNA > poly(C)·poly(G) > tRNAphe > poly(A), (ii) non-cooperative binding of harmalol to poly(C)·poly(G) and poly(A) and cooperative binding with CT DNA and tRNAphe, (iii) significant structural changes of CT DNA, poly(C)·poly(G) and tRNAphe with concomitant induction of optical activity in the bound achiral alkaloid molecules, while with poly(A) no intrinsic CD perturbation was observed, (iv) the binding was predominantly exothermic, enthalpy driven, entropy favoured with CT DNA and poly(C)·poly(G) while it was entropy driven with tRNAphe and poly(A), (v) a hydrophobic contribution and comparatively large role of non-polyelectrolytic forces to Gibbs energy changes with CT DNA, poly(C)·poly(G) and tRNAphe, and (vi) intercalated state of harmalol with CT DNA and poly(C)·poly(G) structure as revealed from molecular docking and supported by the viscometric data. Furthermore, with competition dialysis assay it was shown that harmalol prefers hetero GC sequences. All these findings unequivocally pointed out that harmalol prefers binding with ds CT DNA followed by ds poly(C)·poly(G), clover leaf tRNAphe and least with ss poly(A). The results highlight the importance of structural elements in these natural beta-carboline alkaloids in stabilizing different DNA and RNA of various motifs for developing nucleic acid based better therapeutic agents.Keywords: calorimetry, docking, DNA/RNA-alkaloid interaction, harmalol, spectroscopy
Procedia PDF Downloads 228191 Equipping Organic Farming in Medicinal and Aromatic Plants: Central Institute of Medicinal and Aromatic Plants' Scientific Interventions
Authors: Alok Kalra
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Consumers and practitioners (medical herbalists, pharmacists, and aromatherapists) with strong and increased awareness about health and environment demand organically grown medicinal and aromatic plants (MAPs) to offer a valued product. As the system does not permit the use of synthetic fertilizers the use of nutrient rich organic manures is extremely important. CSIR-CIMAP has developed a complete recycling package for managing distillation and agro-waste of medicinal and aromatic plants for production of superior quality vermicompost involving microbes capable of producing high amounts of humic acid. The major benefits being faster composting period and nutrient rich vermicompost; a nutrient advantage of about 100-150% over the most commonly used organic manure (FYM). At CSIR-CIMAP, strains of microbial inoculants with multiple activities especially strains useful both as biofertilizers and biofungicide and consortia of microbes possessing diverse functional activities have been developed. CSIR-CIMAP has also initiated a program where a large number of accessions are being screened for identifying organic proficient genotypes in mints, ashwagandha, geranium and safed musli. Some of the natural plant growth promoters like calliterpenones from the plant Callicarpa macrophylla has been tested successfully for induction of rooting in stem cuttings and improving growth and yield of various crops. Some of the microbes especially the endophytes have even been identified improving the active constituents of medicinal and aromatic plants. The above said scientific interventions making organic farming a charming proposition would be discussed in details.Keywords: organic agriculture, microbial inoculants, organic fertilizers, natural plant growth promoters
Procedia PDF Downloads 237190 YPFS Attenuating TH2 Cell-Mediated Allergic Inflammation by Regulating the TSLP Pathway
Authors: Xi Yu, Lili Gu, Huizhu Wang, Xiao Wei, Dandan Sheng, Xiaoyan Jiang, Min Hong
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Introduction: Hypersensitivity disease is difficult to cure completely because of its recurrence, yupingfengsan (YPFS) is used to treat the diseases with the advantage of reducing the recurrence,but the precise mechanism is not clear. Previous studies of our laboratory have shown that the extract of YPFS can inhibit Th2-type allergic contact dermatitis(ACD) induced by FITC.Besides, thymic stromal lymphopoietin(TSLP) have been proved to be a master switch for allergic inflammation. Based on these studies, we want to establish a mouse model of TSLP production based on Th2 cell-mediated allergic inflammation to explore the regulating mechanisms of YPFS on TSLP in Th2 cell-mediated allergic inflammation. Methods: Th2-type ACD mouse model: The mice were topically sensitized on the abdomens (induction phase) and elicited on its ears skin 6 day later (excitation phase) with FITC solution, and the ear swelling was measured to evaluate the allergic inflammation;A mouse model of TSLP production based on Th2 cell-mediated allergic inflammation (TSLP production model): the skin of the ear was sensitized on two consecutive days with FITC solution causing the production of TSLP;Mice were treated with YPFS extract,ELISA、Real-time PCR and Western-blotting were using to examine the mRNA and protein levels of TSLP\TSLPR and TLRs ect. Results: YPFS extract can attenuates Th2-type allergic inflammatory in mice;in TSLP production model, YPFS can inhibit the expression of TSLP、 TSLPR、TLRs and MyD88, So we deduce the possible mechanisms of YPFS to play a role of intervention is through TLRs- MyD88 dependent and independent pathway to reduce TSLP production.Keywords: YPFS, TSLP, TLRs, Th2-type allergic contact dermatitis
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