Search results for: secretory proteins
451 Effects of Porcine Oviductal Fluid on In vitro Growth of Dendrobium mirbelianum
Authors: M. Youngsabanant-Areekijseree, C. Thepsithar, K. Sribuddhachart, J. Tananantayot
Abstract:
Porcine oviductal fluid (pOF) from oviduct, an unused organ from the slaughterhouse, was effectively used for biotechnology studies. The fluid components consisted of micro- and macro-nutrients, amino acids, carbon source and proteins that played important roles in animal cell and embryo development. This was our knowledge on investigating pOF as growth promoting substance in culture medium of an orchid, Dendrobium mirbelianum. Two-leaf shoots were cultured in liquid Vacin and Went (VW) medium as a standard medium supplemented with 2 g/L peptone (Pe) or 100 g/ L boiled-potato water (Po) alone or in combinations, and added with 0, 1, 3 or 5 ml/L pOF. All explants were cultured in a stationary condition for 8 weeks. It was found that medium added with 100 g/L Po and 1 ml/L pOF provided the best results (1.02 g fresh weight, 4.2 shoots, 0.53 cm shoot height, 4.4 protocorms, 11.0 leaves and 5.7 roots with 100% survival) when compared to other medium, but not statistically significant difference from medium added with 100 g/L Po (0.86 g fresh weight, 4.3 shoots, 0.51 cm shoot height, 4.6 protocorms, 12.4 leaves and 6.6 roots with 100% survival). However, VW medium supplemented with 1 or 3 ml/L pOF alone showed the higher percentage of survival (100%) than VW medium (86.67%). It was shown the potential role of pOF as an organic supplement for promoting growth of plants. Acknowledgements—The project was funded by a grant from Silpakorn University Research & Development Institute (SURDI) and Faculty of Science, Silpakorn University, Thailand.Keywords: Dendrobium mirbelianum, pig, oviductal fluid, in vitro growth
Procedia PDF Downloads 185450 Helicoverpa armigera Hubner (Lepidoptera: Noctuidae) Susceptibility to Bacillus thuringiensis Crystal Toxins
Authors: Muhammad Jawad Saleem, Faisal Hafeez, Muhammad Arshad, Afifa Naeem, Ayesha Iftekhar
Abstract:
Bacillus thuringiensis is a gram-positive spore-forming bacterium that belongs to the Bacillus cereus group of Bacilli and it produces ICP (insecticidal crystal protein) Cry toxins or Cysts toxins. Spores are produced as parasporal crystalline inclusions bodies (also known as endotoxins) at the onset of sporulation during the stationary growth phase. During vegetative growth that does not form crystals and is called vegetative insecticidal proteins (VIP) and secreted an insecticidal protein (SIP). Bacillus thuringiensis (Bt) is important for pest management either in the form of insecticides or through incorporated in the gene of the crop. Bioassays were conducted on the F2 generation of 1st instar larvae of H. armigera by the diet incorporation method to determine the susceptibility to Bt Cry toxins (Cry1Ac, Cry2Ab, Cry2A). The median lethal concentration (LC₅₀) of Cry1Ac, Cry2Ab, Cry2A ranged from 0.11 to 1.06 µg/ml and moult inhibitory concentration (MIC₅₀) of Cry1Ac, Cry2Ab, Cry2A ranged from 0.05 to 0.25 µg/ml. Cry1Ac was found most toxic to 1st instar larvae of H. armigera as compared to other Bt Cry toxins (Cry1Ac, Cry2Ab, Cry2A). The experimental results are important to policy-makers and technology providers to develop strategies for the exploitation of transgenic Bt cotton varieties as a component of integrated pest management.Keywords: Bt toxin, Cry1Ac, Cry2Ab, Cry2A, susceptibility, Helicoverpa armigera
Procedia PDF Downloads 165449 Characterization of Solanum tuberosum Ammonium Transporter Gene Using Bioinformatics Approach
Authors: Adewole Tomiwa Adetunji, Francis Bayo Lewu, Richard Mundembe
Abstract:
Plants require nitrogen (N) to support desired production levels. There is a need for better understanding of N transport mechanism in order to improve N assimilation by plant root. Nitrogen is available to plants in the form of nitrate or ammonium, which are transported into the cell with the aid of various transport proteins. Ammonium transporters (AMTs) play a role in the uptake of ammonium, the form in which N is preferentially absorbed by plants. Solanum tuberosum AMT1 (StAMT1) was amplified, sequenced and characterized using molecular biology and bioinformatics methods. Nucleotide database sequences were used to design 976 base pairs AMT1-specific primers which include forward primer 5’- GCCATCGCCGCCGCCGG-3’ and reverse primer 5’-GGGTCAGATCCATACCCGC-3’. These primers were used to amplify the Solanum tuberosum AMT1 internal regions. Nucleotide sequencing, alignment and phylogenetic analysis assigned StAMT1 to the AMT1 family due to the clade and high similarity it shared with other plant AMT1 genes. The deduced amino acid sequences showed that StAMT1 is 92%, 83% and 76% similar to Solanum lycopersicum LeAMT1.1, Lotus japonicus LjAMT1.1, and Solanum lycopersicum LeAMT1.2 respectively. StAMT1 fragments were shown to correspond to the 5th-10th trans-membrane domains. Residue StAMT1 D15 is predicted to be essential for ammonium transport, while mutations of StAMT1 S76A may further enhance ammonium transport.Keywords: ammonium transporter, bioinformatics, nitrogen, primers, Solanum tuberosum
Procedia PDF Downloads 220448 Electrophoretic Changes in Testis and Liver of Mice after Exposure to Diclofenac Sodium
Authors: Deepak Mohan, Sushma Sharma, Mohammad Asif
Abstract:
Diclofenac sodium being one of the most common non-steroidal anti-inflammatory drugs is normally used as painkiller and to reduce inflammation. The drug is known to alter the enzymatic activities of acid and alkaline phosphatase, glutamate oxaloacetate transaminase and glutamate pyruvate transaminases. The drug also results in change in the concentration of proteins and lipids in the body. The present study is an attempt to study different biochemical changes electrophoretically due to administration of different doses of diclofenac (4mg/kg/body weight and 14mg/kg/body weight) on liver and testes of mice from 7-28 days of investigation. Homogenization of the tissue was done, supernatant separated was loaded in the gel and native polyacrylamide gel electrophoresis was conducted. Diclofenac administration resulted in alterations of all these biochemical parameters which were observed in native polyacrylamide gel electrophoretic studies. The severe degenerative changes as observed during later stages of the experiment showed correlation with increase or decrease in the activities of all the enzymes studied in the present investigation. Image analysis of gel in liver showed a decline of 7.4 and 5.3 % in low and high dose group after 7 days whereas a decline of 9.6 and 7.5% was registered after 28 days of investigation. Similar analysis for testis also showed an appreciable decline in the activity of alkaline phosphatase after 28 days. Gel analysis of serum was also performed to find a correlation in the enzymatic activities between the tissue and blood.Keywords: diclofenac, inflammation, polyacrylamide, phosphatase
Procedia PDF Downloads 137447 Copper (II) Complex of New Tetradentate Asymmetrical Schiff Base Ligand: Synthesis, Characterization, and Catecholase-Mimetic Activity
Authors: Cahit Demetgul, Sahin Bayraktar, Neslihan Beyazit
Abstract:
Metalloenzymes are enzyme proteins containing metal ions, which are directly bound to the protein or to enzyme-bound nonprotein components. One of the major metalloenzymes that play a key role in oxidation reactions is catechol oxidase, which shows catecholase activity i.e. oxidation of a broad range of catechols to quinones through the four-electron reduction of molecular oxygen to water. Studies on the model compounds mimicking the catecholase activity are very useful and promising for the development of new, more efficient bioinspired catalysts, for in vitro oxidation reactions. In this study, a new tetradentate asymmetrical Schiff-base and its Cu(II) complex were synthesized by condensation of 4-nitro-1,2-phenylenediamine with 6-formyl-7-hydroxy-5-methoxy-2-methylbenzopyran-4-one and by using an appropriate Cu(II) salt, respectively. The prepared compounds were characterized by elemental analysis, FT-IR, NMR, UV-Vis and magnetic susceptibility. The catecholase-mimicking activity of the new Schiff Base Cu(II) complex was performed for the oxidation of 3,5-di-tert-butylcatechol (3,5-DTBC) in methanol at 25 °C, where the electronic spectra were recorded at different time intervals. The yield of the quinone (3,5-DTBQ) was determined from the measured absorbance at 400 nm of the resulting solution. The compatibility of catalytic reaction with Michaelis-Menten kinetics was also investigated. In conclusion, we have found that our new Schiff Base Cu(II) complex presents a significant capacity to catalyze the oxidation reaction of the catechol to o-quinone.Keywords: catecholase activity, Michaelis-Menten kinetics, Schiff base, transition metals
Procedia PDF Downloads 301446 Comparative Survival Rates of Yeasts during Freeze-Drying, Traditional Drying and Spray Drying
Authors: Latifa Hamoudi-Belarbi, L'Hadi Nouri, Khaled Belkacemi
Abstract:
The effect of three methods of drying (traditional drying, freeze-drying and spray-drying) on the survival of concentrated cultures of Geotrichum fragrans and Wickerhamomyces anomalus was studied. The survival of yeast cultures was initially compared immediately after freeze-drying using HES 12%(w/v)+Sucrose 7% (w/v) as protectant, traditional drying in dry rice cakes and finally spray-drying with whey proteins. The survival of G. fragrans and W. anomalus was studied during 4 months of storage at 4°C and 25°C, in the darkness, under vacuum and at 0% relative humidity. The results demonstrated that high survival was obtained using traditional method of preservation in rice cakes (60% for G. fragrans and 65% for W. anomalus) and freeze-drying in (68% for G. fragrans and 74% for W. anomalus). However, poor survival was obtained by spray-drying method in whey protein with 20% for G. fragrans and 29% for W. anomalus. During storage at 25°C, yeast cultures of G. fragrans and W. anomalus preserved by traditional and freeze-drying methods showed no significant loss of viable cells up to 3 months of storage. Spray-dried yeast cultures had the greatest loss of viable count during the 4 months of storage at 25°C. During storage at 4°C, preservation of yeasts cultures using traditional method of preservation provided better survival than freeze-drying. This study demonstrated the effectiveness of the traditional method to preserve yeasts cultures compared to the high cost methods like freeze-drying and spray-drying.Keywords: freeze-drying, traditional drying, spray drying, yeasts
Procedia PDF Downloads 482445 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
Procedia PDF Downloads 152444 Inhibition of Streptococcus Mutans Biofilm Development of Dental Caries In Vitro and In Vivo by Trachyspermum ammi Seeds: An Approach of Alternative Medicine
Authors: Mohd Adil, Rosina Khan, Danishuddin, Asad U. Khan
Abstract:
The aim of this study was to evaluate the influence of the crude and active solvent fraction of Trachyspermum ammi on S. mutans cariogenicity, effect on expression of genes involved in biofilm formation and caries development in rats. GC–MS was carried out to identify the major components present in the crude and the active fraction of T. ammi. The crude extract and the solvent fraction exhibiting least MIC were selected for further experiments. Scanning electron microscopy was carried out to observe the effect of the extracts on S. mutans biofilm. Comparative gene expression analysis was carried out for nine selected genes. 2-Isopropyl-5-methyl-phenol was found as major compound in crude and the active fraction. Binding site of this compound within the proteins involved in biofilm formation was mapped with the help of docking studies. Real-time RT-PCR analyses revealed significant suppression of the genes involved in biofilm formation. All the test groups showed reduction in caries (smooth surface as well as sulcal surface caries) in rats. Moreover, it also provides new insight to understand the mechanism influencing biofilm formation in S. mutans. Furthermore, the data suggest the putative cariostatic properties of T. Ammi and hence can be used as an alternative medicine to prevent caries infection.Keywords: bio-film, Streptococcus mutans, dental caries, bio-informatic
Procedia PDF Downloads 465443 Isolation, Preparation and Biological Properties of Soybean-Flaxseed Protein Co-Precipitates
Authors: Muhammad H. Alu’datt, Inteaz Alli
Abstract:
This study was conducted to prepare and evaluate the biological properties of protein co-precipitates from flaxseed and soybean. Protein was prepared by NaOH extraction through the mixing of soybean flour (Sf) and flaxseed flour (Ff) or mixtures of soybean extract (Se) and flaxseed extract (Fe). The protein co-precipitates were precipitated by isoelectric (IEP) and isoelectric-heating (IEPH) co-precipitation techniques. Effects of extraction and co-precipitation techniques on co-precipitate yield were investigated. Native-PAGE, SDS-PAGE were used to study the molecular characterization. Content and antioxidant activity of extracted free and bound phenolic compounds were evaluated for protein co-precipitates. Removal of free and bound phenolic compounds from protein co-precipitates showed little effects on the electrophoretic behavior of the proteins or the protein subunits of protein co-precipitates. Results showed that he highest protein contents and yield were obtained in for Sf-Ff/IEP co-precipitate with values of 53.28 and 25.58% respectively as compared to protein isolates and other co-precipitates. Results revealed that the Sf-Ff/IEP showed a higher content of bound phenolic compounds (53.49% from total phenolic content) as compared to free phenolic compounds (46.51% from total phenolic content). Antioxidant activities of extracted bound phenolic compounds with and without heat treatment from Sf-Ff/IEHP were higher as compared to free phenolic compounds extracted from other protein co-precipitates (29.68 and 22.84%, respectively).Keywords: antioxidant, phenol, protein co-precipitate, yield
Procedia PDF Downloads 227442 Prospective Study of the Evaluation of Autologous Blood Injection in the Treatment of Lateral Epicondylitis
Authors: Bheeshma B., Mathivanan N., Manoj Deepak M., Prabhu Thangaraju, K. Venkatachalam
Abstract:
This study involves the effect of autologous blood injection for patients who had degeneration of the origin of extensor carpi radialis brevis which was confirmed radio logically and by ultrasound examination and failed cortisone injections to the lateral epicondylitis. In this prospective longitudinal series involves pre-injection assessment of grip strength, pain, and function, using the patient-rated tennis elbow evaluation. In this study, blood from the contralateral limb is taken and injected into the affected limb with the help of ultrasound guidance and then the patient wore a customized wrist support for five days, after which they were commenced with stretching, strengthening, and massage programme with an occupational therapist. In these patients assessment was done after six months and then finally at 12 months after injection, using the patient-rated tennis elbow evaluation. 50 patients completed the study, showing significant improvement in pain; the worst pain decreased by two to five points out of a 10-point visual analogue for pain. Self-perceived function improved by 11–25 points out of 100. Women showed significant increase in grip, but men did not. Our study thus concludes that autologous blood injection show significant improvement in pain and function in patients with chronic lateral epicondylitis, who did not have relief with cortisone injection.Keywords: lateral epicondylitis, autologous blood injection, conservative treatment, plasma-rich proteins (PRPs)
Procedia PDF Downloads 423441 Anti-Nutritional Factors, In-Vitro Trypsin, Chymotrypsin and Peptidase Multi Enzyme Protein Digestibility of Some Melon (Egusi) Seeds and Their Protein Isolates
Authors: Joan O. Ogundele, Aladesanmi A. Oshodi, Adekunle I. Amoo
Abstract:
Abstract In-vitro multi-enzyme protein digestibility (IVMPD) and some anti-nutritional factors (ANF) of five melon (egusi) seed flours (MSF) and their protein isolates (PI) were carried out. Their PI have potentials comparable to that of soya beans. It is important to know the IVMPD and ANF of these protein sources as to ensure their safety when adapted for use as alternate protein sources to substitute for cow milk, which is relatively expensive in Nigeria. Standard methods were used to produce PI of Citrullus colocynthis, Citrullus vulgaris, African Wine Kettle gourd (Lageneria siceraria I), Basket Ball gourd (Lagenaria siceraria II) and Bushel Giant Gourd (Lageneria siceraria III) seeds and to determine the ANF and IVMPD of the MSF and PI unheated and at 37oC. Multi-enzymes used were trypsin, chymotrypsin and peptidase. IVMPD of MSF ranged from (70.67±0.70) % (C. vulgaris) to (72.07± 1.79) % (L.siceraria I) while for their PI ranged from 74.33% (C.vulgaris) to 77.55% (L.siceraria III). IVMPD of the PI were higher than those of MSF. Heating increased IVMPD of MSF with average value of 79.40% and those of PI with average of 84.14%. ANF average in MSF are tannin (0.11mg/g), phytate (0.23%). Differences in IVMPD of MSF and their PI at different temperatures may arise from processing conditions that alter the release of amino acids from proteins by enzymatic processes. ANF in MSF were relatively low, but were found to be lower in the PI, therefor making the PI safer for human consumption as an alternate source of protein.Keywords: Anti-nutrients, Enzymatic protein digestibility, Melon (egusi)., Protein Isolates.
Procedia PDF Downloads 107440 Therapeutic Evaluation of Bacopa Monnieri Extract on Liver Fibrosis in Rats
Authors: Yu Wen Wang, Shyh Ming Kuo, Hsia Ying Cheng, Yu Chiuan Wu
Abstract:
Liver fibrosis is caused by the activation of hepatic stellate cells in the liver to secrete excessive and deposition of extracellular matrix. In recent years, many treatment strategies have been developed to reduce the activation of hepatic stellate cells and therefore to increase the decomposition of extracellular matrix. Bacopa monnieri, an herbaceous plant of the scrophulariaceae, containing saponins and glycosides, which with antioxidant, anti-inflammation, pain relief and free radical scavenging characteristics. This study was to evaluate the inhibition of hepatic stellate cell activity by Bacopa monnieri extract and its therapeutic potential in treating thioacetamide-induced liver fibrosis in rats. The results showed that the IC50 of Bacopa monnieri extract was 0.39 mg/mL. Bacopa monnieri extract could effectively reduce H2O2-induced hepatic stellate cells inflammation. In the TAA-induced liver fibrosis animal studies, albumin secretion recovered to normal level after treated with Bacopa monnieri extract for 2-w, and fibrosis related proteins, α-SMA and TGF-1levels decreased indicating the extract exerted therapeutic effect on the liver fibrosis. However, inflammatory factors TNF- obviously decreased after 4-w treatment. In summary, we could successfully extract the main component-Bacopaside I from the plant and acquired a potential therapy using this component in treating TAA-induced liver fibrosis in rat.Keywords: anti-inflammatory, Bacopa monnieri, fibrosis, hepatic stellate cells, water extract
Procedia PDF Downloads 98439 Protective Role of Phycobiliproteins in ROS-Associated Physiological Anomalies
Authors: Ravi Raghav Sonani, Niraj Kumar Singh, Jitendra Kumar, Datta Madamwar
Abstract:
Phycobiliproteins (PBPs) are light harvesting proteins showing very strong absorbance and fluorescence in the visible range of the solar spectrum. Phycoerythrin (PE) and phycocyanin (PC) are majorly found PBPs in the cyanobacteria and red algae. In the present study, we have investigated the reactive oxygen species (ROS)-averting capacity of purified PE and PC of cyanobacterial origin. Furthermore, the possibility - whether the ROS-averting potential of PBPs can be explored in the therapeutics of oxidative stress associated physiological anomalies including aging and neurodegenerative diseases. The nematode Caenorhabditis elegans has been used as model organism in this study. PE and PC treatment moderated normal aging and associated physiological functionalities like pharyngeal pumping and locomotion of C. elegans. Moreover, PE-treatment enhanced the stress (oxidative and heat) tolerance upon PE and PC treatment. Specifically, PE treatment was also noted to moderate the progression of Alzheimer’s disease in transgenic C. elegans CL4176. However, PC-treatment curtailed the polyQ aggregation mediated proteotoxicity in C. elegans AM141 (Huntington disease model) under stressed (paraquat stress) as well as normal conditions. The effectiveness of PE and PC in expanding the lifespan of mutant C. elegans knockout for some up- (daf 16) and down- (daf-2 and age-1) stream regulators of insulin/IGF-1 signalling (IIS) shows the independency of their effects from DAF-2–AGE-1–DAF-16 signalling pathway. In conclusion, the present report demonstrates the anti-aging and neuro-protective potential of cyanobacterial PE and PC.Keywords: phycobiliproteins, aging, alzheimer, huntington, C. elegans
Procedia PDF Downloads 384438 Efficiency of Natural Metabolites on Quality Milk Production in Mixed Breed Cows.
Authors: Mariam Azam, Sajjad Ur Rahman, Mukarram Bashir, Muhammad Tahir, Seemal Javaid, Jawad, Aoun Muhammad, Muhammad Zohaib, Hannan Khan
Abstract:
Products of microbial origin are of great importance as they have proved their value in healthcare and nutrition, use of these microbial metabolites acquired from partially fermented soya hulls and wheat bran along with Saccharomyces cerevisiae (DL-22 S/N) substantiates to be a great source for an increase in the total milk production and quality yield.1×109 CFU/ml cells of Saccharomyces cerevisiae (DL-22 S/N) were further grown under in-vivo conditions for the assessment of quality milk production. Two groups with twelve cows, each having the same physical characteristics (Group A and Group B), were under study, Group A was daily fed with 12gm of biological metabolites and 22% protein-pelleted feed. On the other hand, the animals of Group B were provided with no metabolites in their feed. In thirty days of trial, improvement in the overall health, body score, milk protein, milk fat, yield, incidence rate of mastitis, ash, and solid not fat (SNF) was observed. The collected data showed that the average quality milk production was elevated up to 0.45 liter/h/d. However, a reduction in the milk fats up to 0.45% and uplift in the SNF value up to 0.53% of cow milk was also observed. At the same time, the incidence rate of mastitis recorded for the animals under trial was reduced to half, and improved non specific immunity was reported.Keywords: microbial metabolites, post-biotics, animal supplements, animal nutrition, proteins, animal production, fermentation
Procedia PDF Downloads 85437 Cloning and Functional Analysis of NtPIN1a Promoter Under Various Abiotic Stresses in Nicotiana Tabacum
Authors: Zia Ullah, Muhammad Asim, Shi Sujuan, Rayyan Khan, Aaqib Shaheen, LIU Haobao
Abstract:
The plant-specific auxin efflux proteins PIN-FORMED (PIN) have been well depicted in many plant species for their essential roles in regulating the transport of auxins in several phases of plant growth. Little is known about the various functions of the PIN family genes in the Nicotiana tabacum (N. tabacum) species during plant growth. To define the expression pattern of the NtPIN1a gene under abiotic stresses and hormone treatment, transgenic tobacco with promoterNtPIN1a::GUS construct was employed. Comprehensive computational analyses of the NtPIN1a promoter confirmed the existence of common core promoter elements including CAAT-box, TATA-box, hormone, and abiotic stress-responsive elements such as ABRE, P-box, MYC, MYB, ARE, and GC-motifs. The transgenic plants with the promoter of NtPIN1a displayed a promising expression of β-glucuronidase (GUS) in germinating seeds, root tips, shoot-apex, and developing leaves under optimal conditions. While the differential expression of GUS in moderate salt, drought, low potassium stresses, and externally high auxin level at two different time points, suggested NtPIN1a played a key role in growth processes and the plants’ response to abiotic stresses. This analysis provides a foundation for more in-depth discoveries of the biological functions of NtPIN1a in Nicotiana species and this promoter may be employed in genetic engineering of other crops for enhanced stress tolerance.Keywords: tobacco, nicotiana tabacum, pin, promoter, GUS, abiotic stresses, auxin
Procedia PDF Downloads 85436 The Role Collagen VI Plays in Heart Failure: A Tale Untold
Authors: Summer Hassan, David Crossman
Abstract:
Myocardial fibrosis (MF) has been loosely defined as the process occurring in the pathological remodeling of the myocardium due to excessive production and deposition of extracellular matrix (ECM) proteins, including collagen. This reduces tissue compliance and accelerates progression to heart failure, as well as affecting the electrical properties of the myocytes resulting in arrhythmias. Microscopic interrogation of MF is key to understanding the molecular orchestrators of disease. It is well-established that recruitment and stimulation of myofibroblasts result in Collagen deposition and the resulting expansion in the ECM. Many types of Collagens have been identified and implicated in scarring of tissue. In a series of experiments conducted at our lab, we aim to elucidate the role collagen VI plays in the development of myocardial fibrosis and its direct impact on myocardial function. This was investigated through an animal experiment in Rats with Collagen VI knockout diseased and healthy animals as well as Collagen VI wild diseased and healthy rats. Echocardiogram assessments of these rats ensued at four-time points, followed by microscopic interrogation of the myocardium aiming to correlate the role collagen VI plays in myocardial function. Our results demonstrate a deterioration in cardiac function as represented by the ejection fraction in the knockout healthy and diseased rats. This elucidates a potential protective role that collagen-VI plays following a myocardial insult. Current work is dedicated to the microscopic characterisation of the fibrotic process in all rat groups, with the results to follow.Keywords: heart failure, myocardial fibrosis, collagen, echocardiogram, confocal microscopy
Procedia PDF Downloads 70435 Influence of Cooking on the Functional Properties of Dioscorea Schimperiana During Chips Production
Authors: Djeukeu Asongni William, Leng Marlyse, Gouado Inocent
Abstract:
Background: Process for obtaining D. schimperiana chips involves a long period of cooking followed by drying of obtained products in the sun. Such a process could induce the modification of the functional properties of the chips, thus reducing the technological uses of these products. This study was conducted with a view to assessing the impact of this process on the chips of D. schimperiana. Methods: The chips used were purchased in Baham, Bamendjou and Bagangté markets during the month of February 2013. A representative sample of each market chips was formed by mixing the chips of several sellers. The control sample consisted of fresh yams that have been sliced to the average size of local chips then dried in the oven at 45 ° C for 36 h. On each sample was performed the analysis of the physico-chemical properties (carbohydrates, lipids, proteins, iron , phosphorus, reducing sugars, ash and total starch) and gelling properties both with and without inhibitor alpha-amylases (0.018 and 0.146 mol / l). Results: Results show that the levels of ash 2.99 g / 100gms, iron 1.01 g / 100gms and phosphorus 532.06 mg / 100gms fresh sample were significantly higher than those of the products obtained in the traditional process. The functional properties of the chips obtained from different methods shows that the peak viscosity of the fresh sample is larger than the other samples with or without inhibitor. In addition, the fresh sample has the lowest breakdown under the same conditions. Conclusion: These results show that traditional process reduces technological potential of chips, thus limiting the value of D. schimperiana.Keywords: Dioscorea schimperiana, chips, functional properties, technological properties, valorization
Procedia PDF Downloads 392434 Comprehensive Multi-Omics Study Highlights Osteopontin/SPP1 in Ovarian Aging Control
Authors: Chia-Jung Li, Li-Te Lin, Kuan-Hao Tsui
Abstract:
The study identifies SPP1 as a potential gene associated with ovarian aging, revealing a significant decline in its expression in aged ovaries. SPP1, also known as osteopontin (OPN), is a multifunctional glycoprotein involved with regulatory proteins and pro-inflammatory immune chemokines. However, its genetic links to ovarian aging have not been extensively explored. Spatial transcriptomic analyses were conducted on ovaries from young and aged female mice, along with a sample from a 73-year-old individual. Additionally, single-cell RNA sequencing analysis was performed to identify associations between SPP1 and key genes. The study focused on crucial genes, including ITGAV, ITGB1, CD44, MMP3, and FN1, with a particular emphasis on the correlation between SPP1 and ITGB1. The findings indicate a significant decline in SPP1 expression in aged ovaries, which was consistent in the 73-year-old sample. Single-cell RNA sequencing unveiled associations between SPP1 and key genes, emphasizing a strong co-expression correlation between SPP1 and ITGB1. While the study provides valuable insights, further research is necessary to understand the broader implications and potential applications of SPP1 in ovarian aging. Translating these findings to clinical settings requires careful consideration. The identification of SPP1 as a gene implicated in ovarian aging opens new avenues for advancing precision medicine and refining treatment strategies for conditions related to ovarian aging.Keywords: SPP1, ovarian aging, spatial transcriptomic, single-cell RNA sequencing
Procedia PDF Downloads 27433 The Effect of Particulate Matter on Cardiomyocyte Apoptosis Through Mitochondrial Fission
Authors: Tsai-chun Lai, Szu-ju Fu, Tzu-lin Lee, Yuh-Lien Chen
Abstract:
There is much evidence that exposure to fine particulate matter (PM) from air pollution increases the risk of cardiovascular morbidity and mortality. According to previous reports, PM in the air enters the respiratory tract, contacts the alveoli, and enters the blood circulation, leading to the progression of cardiovascular disease. PM pollution may also lead to cardiometabolic disturbances, increasing the risk of cardiovascular disease. The effects of PM on cardiac function and mitochondrial damage are currently unknown. We used mice and rat cardiomyocytes (H9c2) as animal and in vitro cell models, respectively, to simulate an air pollution environment using PM. These results indicate that the apoptosis-related factor PUMA, a regulator of apoptosis upregulated by p53, is increased in mice treated with PM. Apoptosis was aggravated in cardiomyocytes treated with PM, as measured by TUNEL assay and Annexin V/PI. Western blot results showed that CASPASE3 was significantly increased and BCL2 (B-cell lymphoid 2) was significantly decreased under PM treatment. Concurrent exposure to PM increases mitochondrial reactive oxygen species (ROS) production by MitoSOX Red staining. Furthermore, using Mitotracker staining, PM treatment significantly shortened mitochondrial length, indicating mitochondrial fission. The expression of mitochondrial fission-related proteins p-DRP1 (phosphodynamics-related protein 1) and FIS1 (mitochondrial fission 1 protein) was significantly increased. Based on these results, the exposure to PM worsens mitochondrial function and leads to cardiomyocyte apoptosis.Keywords: particulate matter, cardiomyocyte, apoptosis, mitochondria
Procedia PDF Downloads 93432 Efficient Production of Cell-Adhesive Motif From Human Fibronectin Domains to Design a Bio-Functionalized Scaffold for Tissue Engineering
Authors: Amina Ben Abla, Sylvie Changotade, Geraldine Rohman, Guilhem Boeuf, Cyrine Dridi, Ahmed Elmarjou, Florence Dufour, Didier Lutomski, Abdellatif Elm’semi
Abstract:
Understanding cell adhesion and interaction with the extracellular matrix is essential for biomedical and biotechnological applications, including the development of biomaterials. In recent years, numerous biomaterials have emerged and were used in the field of tissue engineering. Nevertheless, the lack of interaction of biomaterials with cells still limits their bio-integration. Thus, the design of bioactive biomaterials to improve cell attachment and proliferation is of growing interest. In this study, bio-functionalized material was developed combining a synthetic polymer scaffold surface with selected domains of type III human fibronectin (FNIII-DOM) to promote cell adhesion and proliferation. Bioadhesive ligand includes cell-binding domains of human fibronectin, a major ECM protein that interacts with a variety of integrins cell-surface receptors, and ECM proteins through specific binding domains were engineered. FNIII-DOM was produced in bacterial system E. coli in 5L fermentor with a high yield level reaching 20mg/L. Bioactivity of the produced fragment was validated by studying cellular adhesion of human cells. The adsorption and immobilization of FNIII-DOM onto the polymer scaffold were evaluated in order to develop an innovative biomaterial.Keywords: biomaterials, cellular adhesion, fibronectin, tissue engineering
Procedia PDF Downloads 136431 Insight into Structure and Functions of of Acyl CoA Binding Protein of Leishmania major
Authors: Rohit Singh Dangi, Ravi Kant Pal, Monica Sundd
Abstract:
Acyl-CoA binding protein (ACBP) is a housekeeping protein which functions as an intracellular carrier of acyl-CoA esters. Given the fact that the amastigote stage (blood stage) of Leishmania depends largely on fatty acids as the energy source, of which a large part is derived from its host, these proteins might have an important role in its survival. In Leishmania major, genome sequencing suggests the presence of six ACBPs, whose function remains largely unknown. For functional and structural characterization, one of the ACBP genes was cloned, and the protein was expressed and purified heterologously. Acyl-CoA ester binding and stoichiometry were analyzed by isothermal titration calorimetry and Dynamic light scattering. Our results shed light on high affinity of ACBP towards longer acyl-CoA esters, such as myristoyl-CoA to arachidonoyl-CoA with single binding site. To understand the binding mechanism & dynamics, Nuclear magnetic resonance assignments of this protein are being done. The protein's crystal structure was determined at 1.5Å resolution and revealed a classical topology for ACBP, containing four alpha-helical bundles. In the binding pocket, the loop between the first and the second helix (16 – 26AA) is four residues longer from other extensively studied ACBPs (PfACBP) and it curls upwards towards the pantothenate moiety of CoA to provide a large tunnel space for long acyl chain insertion.Keywords: acyl-coa binding protein (ACBP), acyl-coa esters, crystal structure, isothermal titration, calorimetry, Leishmania
Procedia PDF Downloads 443430 Efficient Delivery of Biomaterials into Living Organism by Using Noble Metal Nanowire Injector
Authors: Kkochorong Park, Keun Cheon Kim, Hyoban Lee, Eun Ju Lee, Bongsoo Kim
Abstract:
Introduction of biomaterials such as DNA, RNA, proteins is important for many research areas. There are many methods to introduce biomaterials into living organisms like tissue and cells. To introduce biomaterials, several indirect methods including virus‐mediated delivery, chemical reagent (i.e., lipofectamine), electrophoresis have been used. Such methods are passive delivery using an endocytosis process of cell, reducing an efficiency of delivery. Unlike the indirect delivery method, it has been reported that a direct delivery of exogenous biomolecules into nucleus have been more efficient to expression or integration of biomolecules. Nano-sized material is beneficial for detect signal from cell or deliver stimuli/materials into the cell at cellular and molecular levels, due to its similar physical scale. Especially, because 1 dimensional (1D) nanomaterials such as nanotube, nanorod and nanowire with high‐aspect ratio have nanoscale geometry and excellent mechanical, electrical, and chemical properties, they could play an important role in molecular and cellular biology. In this study, by using single crystalline 1D noble metal nanowire, we fabricated nano-sized 1D injector which can successfully interface with living cells and directly deliver biomolecules into several types of cell line (i.e., stem cell, mammalian embryo) without inducing detrimental damages on living cell. This nano-bio technology could be a promising and robust tool for introducing exogenous biomaterials into living organism.Keywords: DNA, gene delivery, nanoinjector, nanowire
Procedia PDF Downloads 264429 Colour Formation and Maillard Reactions in Spray-Dried Milk Powders
Authors: Zelin Zhou, Timothy Langrish
Abstract:
Spray drying is the final stage of milk powder production. Traditionally, the quality of spray-dried milk powders has mainly been assessed using their physical properties, such as their moisture contents, while chemical changes occurring during the spray drying process have often been ignored. With growing concerns about food quality, it is necessary to establish a better understanding of heat-induced degradation due to the spray-drying process of skim milk. In this study, the extent of thermal degradation for skim milk in a pilot-scale spray dryer has been investigated using different inlet gas temperatures. The extent of heat-induced damage has been measured by the formation of advanced Maillard reaction products and the loss of soluble proteins at pH 4.6 as assessed by a fluorometric method. A significant increase in the extent of thermal degradation has been found when the inlet gas temperature increased from 170°C to 190°C, suggesting protein unfolding may play an important role in the kinetics of heat-induced degradation for milk in spray dryers. Colour changes of the spray-dried skim milk powders have also been analysed using a standard lighting box. Colourimetric analysis results were expressed in CIELAB colour space with the use of the E index (E) and the Chroma (C) for measuring the difference between colours and the intensity of the colours. A strong linear correlation between the colour intensity of the spray-dried skim milk powders and the formation of advanced Maillard reaction products has been observed.Keywords: colour formation, Maillard reactions, spray drying, skim milk powder
Procedia PDF Downloads 172428 Effect of Brewing on the Bioactive Compounds of Coffee
Authors: Ceyda Dadali, Yeşim Elmaci
Abstract:
Coffee was introduced as an economic crop during the fifteenth century; nowadays it is the most important food commodity ranking second after crude oil. Desirable sensory properties make coffee one of the most often consumed and most popular beverages in the world. The coffee preparation method has a significant effect on flavor and composition of coffee brews. Three different extraction methodologies namely decoction, infusion and pressure methods have been used for coffee brew preparation. Each of these methods is related to specific granulation (coffee grind) of coffee powder, water-coffee ratio temperature and brewing time. Coffee is a mixture of 1500 chemical compounds. Chemical composition of coffee highly depends on brewing methods, coffee bean species and roasting time-temperature. Coffee contains a wide number of very important bioactive compounds, such as diterpenes: cafestol and kahweol, alkaloids: caffeine, theobromine and trigonelline, melanoidins, phenolic compounds. The phenolic compounds of coffee include chlorogenic acids (quinyl esters of hidroxycinnamic acids), caffeic, ferulic, p-coumaric acid. In coffee caffeoylquinic acids, feruloylquinic acids and di-caffeoylquinic acids are three main groups of chlorogenic acids constitues 6% -10% of dry weight of coffee. The bioavailability of chlorogenic acids in coffee depends on the absorption and metabolization to biomarkers in individuals. Also, the interaction of coffee polyphenols with other compounds such as dietary proteins affects the biomarkers. Since bioactive composition of coffee depends on brewing methods effect of coffee brewing method on bioactive compounds of coffee will be discussed in this study.Keywords: bioactive compounds of coffee, biomarkers, coffee brew, effect of brewing
Procedia PDF Downloads 188427 Application of Biosensors in Forensic Analysis
Authors: Shirin jalili, Hadi Shirzad, Samaneh Nabavi, Somayeh Khanjani
Abstract:
Biosensors in forensic analysis are ideal biological tools that can be used for rapid and sensitive initial screening and testing to detect of suspicious components like biological and chemical agent in crime scenes. The wide use of different biomolecules such as proteins, nucleic acids, microorganisms, antibodies and enzymes makes it possible. These biosensors have great advantages such as rapidity, little sample manipulation and high sensitivity, also Because of their stability, specificity and low cost they have become a very important tool to Forensic analysis and detection of crime. In crime scenes different substances such as rape samples, Semen, saliva fingerprints and blood samples, act as a detecting elements for biosensors. On the other hand, successful fluid recovery via biosensor has the propensity to yield a highly valuable source of genetic material, which is important in finding the suspect. Although current biological fluid testing techniques are impaired for identification of body fluids. But these methods have disadvantages. For example if they are to be used simultaneously, Often give false positive result. These limitations can negatively result the output of a case through missed or misinterpreted evidence. The use of biosensor enable criminal researchers the highly sensitive and non-destructive detection of biological fluid through interaction with several fluid-endogenous and other biological and chemical contamination at the crime scene. For this reason, using of the biosensors for detecting the biological fluid found at the crime scenes which play an important role in identifying the suspect and solving the criminal.Keywords: biosensors, forensic analysis, biological fluid, crime detection
Procedia PDF Downloads 1101426 Role of Surfactant Protein D (SP-D) as a Biomarker of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) Infection
Authors: Lucia Salvioni, Pietro Giorgio Lovaglio, Valerio Leoni, Miriam Colombo, Luisa Fiandra
Abstract:
The involvement of plasmatic surfactant protein-D (SP-D) in pulmonary diseases has been long investigated, and over the last two years, more interest has been directed to determine its role as a marker of COVID-19. In this direction, several studies aimed to correlate pulmonary surfactant proteins with the clinical manifestations of the virus indicated SP-D as a prognostic biomarker of COVID-19 pneumonia severity. The present work has performed a retrospective study on a relatively large cohort of patients of Hospital Pio XI of Desio (Lombardia, Italy) with the aim to assess differences in the hematic SP-D concentrations among COVID-19 patients and healthy donors and the role of SP-D as a prognostic marker of severity and/or of mortality risk. The obtained results showed a significant difference in the mean of log SP-D levels between COVID-19 patients and healthy donors, so as between dead and survived patients. SP-D values were significantly higher for both hospitalized COVID-19 and dead patients, with threshold values of 150 and 250 ng/mL, respectively. SP-D levels at admission and increasing differences among follow-up and admission values resulted in the strongest significant risk factors of mortality. Therefore, this study demonstrated the role of SP-D as a predictive marker of SARS-CoV-2 infection and its outcome. A significant correlation of SP-D with patient mortality indicated that it is also a prognostic factor in terms of mortality, and its early detection should be considered to design adequate preventive treatments for COVID-19 patients.Keywords: SARS-CoV-2 infection, COVID-19, surfactant protein-D (SP-D), mortality, biomarker
Procedia PDF Downloads 67425 Effect of Fat Percentage and Prebiotic Composition on Proteolysis, ACE-Inhibitory and Antioxidant Activity of Probiotic Yogurt
Authors: Mohammad B. HabibiNajafi, Saeideh Sadat Fatemizadeh, Maryam Tavakoli
Abstract:
In recent years, the consumption of functional foods, including foods containing probiotic bacteria, has come to notice. Milk proteins have been identified as a source of angiotensin-I-converting enzyme )ACE( inhibitory peptides and are currently the best-known class of bioactive peptides. In this study, the effects of adding prebiotic ingredients (inulin and wheat fiber) and fat percentage (0%, 2% and 3.5%) in yogurt containing probiotic Lactobacillus casei on physicochemical properties, degree of proteolysis, antioxidant and ACE-inhibitory activity within 21 days of storage at 5 ± 1 °C were evaluated. The results of statistical analysis showed that the application of prebiotic compounds led to a significant increase in water holding capacity, proteolysis and ACE-inhibitory of samples. The degree of proteolysis in yogurt increases as storage time elapses (P < 0.05) but when proteolysis exceeds a certain threshold, this trend begins to decline. Also, during storage time, water holding capacity reduced initially but increased thereafter. Moreover, based on our findings, the survival of Lactobacillus casei in samples treated with inulin and wheat fiber increased significantly in comparison to the control sample (P < 0.05) whereas the effect of fat percentage on the survival of probiotic bacteria was not significant (P = 0.095). Furthermore, the effect of prebiotic ingredients and the presence of probiotic cultures on the antioxidant activity of samples was significant (P < 0.05).Keywords: probiotic yogurt, proteolysis, ACE-inhibitory, antioxidant activity
Procedia PDF Downloads 240424 Conserved Stem-Loop Structure at the End of Short Interspersed Nuclear Elements (SINE) and Long Interspersed Nuclear Elements (LINE) Pairs of Different Species
Authors: Daria Grechishnikova, Maria Poptsova
Abstract:
Transposable elements play an important role in the evolution of various species from bacteria to human. Long Interspersed Nuclear Elements (LINEs) and Short Interspersed Nuclear Elements (SINEs) are two major classes of retrotransposons that occupy a considerable part of any genome and their copy numbers can range form several hundreds to a million. Both LINEs and SINEs multiply through a copy-and-paste mechanism. LINEs encode proteins, which make them capable of self-propagation while SINEs are parasitic and require the machinery of LINEs to multiply. The mechanisms how LINE and SINE RNA is recognized by the LINE-encoded reverse transcriptase (RT) remain unclear. For some SINE-LINE pairs, it was shown that they share a common 3’-end with a stem-loop structure. Majority of the SINE-LINE pairs do not have a common 3’-end. Recently we have shown that in the human genome Alu-L1 pairs have structurally similar stem-loop structure at the 3’-end. Here we extended our analysis to a wide range of species and analyzed LINEs from 161 different species from Repbase and 217 SINE sequences from SINEBase. It appeared that all of the analyzed sequences contained stem-loop structures at the 3’-end. Here we conclude that it is very likely that a common evolutionary mechanism of transposon RNA recognition requires the presence of stem-loop structures at their 3’-end.Keywords: LINE, SINE, mechanisms of retrotransposition, retrotransposons, stem-loop, stem-loop structures, transposons
Procedia PDF Downloads 343423 Functional Characteristics of Chemosensory Proteins in the Sawyer Beetle Monochamus alternatus Hope
Authors: Saqib Ali, Man-Qun Wang
Abstract:
The Japanese pine sawyer, Monochamus alternatus Hope (Coleoptera: Cerambycidae), is a major pest of pines and it is also the key vector of the exotic pinewood nematode in China. In the present study, we cloned, expressed, and purified a chemosensory protein (CSP) in M. alternatus. We surveyed its expression in various developmental stages of male and female adult tissues and determined its binding affinities for different pine volatiles using a competitive binding fluorescence assay. A CSP known as CSP5 in M. alternatus was obtained from an antennal cDNA library and expressed in Escherichia coli. Quantitative reverse transcription polymerase chain reaction results indicated that the CSP5 gene was mainly expressed in male and female antennae. Competitive binding assays were performed to test the binding affinity of recombinant CSP5 to 13 odour molecules of pine volatiles. The results showed that CSP5 showed very strong binding abilities to myrcene, (+)-β-pinene, and (−)-isolongifolene, whereas the volatiles 2-methoxy-4-vinylphenol, p-cymene, and (+)-limonene oxide have relatively weak binding affinity at pH 5.0. Three volatiles myrcene, (+)-β-pinene, and (−)-isolongifolene may play crucial roles in CSP5 binding with ligands, but this needs further study for confirmation. The sensitivity of insect to host plant volatiles can effectively be used to control and monitor the population through mass trapping as part of integrated pest management programs.Keywords: olfactory-specific protein, volatiles, competitive binding assay, expression characteristics, qPCR
Procedia PDF Downloads 122422 Drug-Drug Plasma Protein Binding Interactions of Ivacaftor
Authors: Elena K. Schneider, Johnny X. Huang, Vincenzo Carbone, Mark Baker, Mohammad A. K. Azad, Matthew A. Cooper, Jian Li, Tony Velkov
Abstract:
Ivacaftor is a novel CF trans-membrane conductance regulator (CFTR) potentiator that improves the pulmonary function for cystic fibrosis patients bearing a G551D CFTR-protein mutation. Because ivacaftor is highly bound (>97%) to plasma proteins, there is the strong possibility that co-administered CF drugs that compete for the same plasma protein binding sites and impact the free drug concentration. This in turn could lead to drastic changes in the in vivo efficacy of ivacaftor and therapeutic outcomes. This study compares the binding affinity of ivacaftor and co-administered CF drugs for human serum albumin (HSA) and α1-acid glycoprotein (AGP) using surface plasmon resonance and fluorimetric binding assays that measure the displacement of site selective probes. Due to their high plasma protein binding affinities, drug-drug interactions between ivacaftor are to be expected with ducosate, montelukast, ibuprofen, dicloxacillin, omeprazole and loratadine. The significance of these drug-drug interactions is interpreted in terms of the pharmacodynamic/pharmacokinetic parameters and molecular docking simulations. The translational outcomes of the data are presented as recommendations for a staggered treatment regimen for future clinical trials which aims to maximize the effective free drug concentration and clinical efficacy of ivacaftor.Keywords: human α-1-acid glycoprotein, binding affinity, human serum albumin, ivacaftor, cystic fibrosis
Procedia PDF Downloads 301