Day 2 :
Biosynthesis Inc., Lewisville, Texas, USA
Keynote: Amino Acid Analysis used in Industry
Time : 10:00-10:40
Dr. Klaus D. Linse has earned a Ph.D. in Cell Biology/Biochemistry at the University of Hamburg, Germany, investigating microtubule proteins at the Max-Planck-Institute for Medical Research in Heidelberg and at DESY Hamburg. He is currently a Director of Scientific Operations at Biosynthesis Inc. in Lewisville, Texas. After receiving his Ph.D., he held positions at Porton Instruments Inc., and Beckman Instruments in instrument development and support. In 1944 he accepted position as Core Facility Director at the University of Texas at Austin. From 2009 to 2012 he was employed as a senior research scientist II at XBiotech Inc., Austin, Texas, in mass spectrometry analyzing therapeutic monoclonal antibodies.
Amino Acid Analysis (AAA) of free amino acids allow for the analysis of raw materials such as food stuff, neutraceuticals, supplements, effervescent tablets, animal feeds, and even blood samples for newborn screening to allow detection of inborn metabolic errors. Recent renewed interest in metabolomic studies has increased the need for sensitive and accurate methods for the analysis of physiological amino acids found in biological samples such as blood, serum, urine as well as neurons. AAA allows for the quantitative analysis of compounds containing primary and secondary amino groups. AAA has become the gold standard for quantitative analysis of free amino acids as well as amino acids released from peptides and proteins via hydrolysis. Amino acid analysis was introduced as an analytical technique in 1959 by Moore and Stein. Since then many improvements in instrument development and design have been made, including the GC-MS and LC-MS/MS-based methods. As instrument and column technology improved, several automated or semi-automated methods for amino analysis have been developed. Two types of analysis methods emerged for automated amino acids: (1) Post-column derivatization, and (2) pre-column derivatization based amino acid analysis. Over the years, several UV-absorbing and fluorescent derivatization compounds have been investigated and are now available for pre-column derivatization. However, for post-column derivatization, ninhydrin is the sole derivatization compound used. To this day, amino acid analysis has remained the gold standard for amino acid analysis. However, for dietary supplements there appears to be presently no standard method available. A brief history of amino acid analysis, as well as instrumentation and methods used, will be discussed.
- Sessions: Lipid Health and Nutrition |Xenobiotics and Endocrinology | Animal Biochemistry
Mississippi Valley State University, USA
Title: Investigating the role of metal enzymes in biosynthesis of protein and fatty acids by application of trace elements
Time : 10:40-11:10
Goli Mudlagiri B has done his PhD in Chemistry. He is teaching economically challenged poor neighborhood kids of delta, Mississippi. His academic interest is in multi-disciplinary areas: Nuclear power plant and in Plutonium plant (BARC, India), other areas of expertise are organic synthesis/ organo-metallic, analytical, medicinal chemistry, nuclear waste disposal. He had the opportunity to discover a novel method for removing gadolinium from heavy water, Aiken, SC.
The present study is part of our ongoing investigation to study the role of trace elements on the effect on soybean seed composition. This was conducted to study the effects of five trace elements. (Mn, Cu, Zn, Mo and B (mainly chlorides except Mo as oxide, and B as boric acid)) alone and in combination with the chelating agent citric acid (CA) on soybean seed protein, oil, and fatty acids soybean cultivar (Bolivar with maturity group V) was grown in a repeated greenhouse experiment in a randomized complete block design. The compounds were applied in a combination of the chelating agent CA and the trace element (example, Mn+CA) to three- week-old soybean plants at V3 (vegetative) and at R3 (beginning of seed pod initiation) stages. The plants were allowed to grow until maturity under greenhouse conditions. The harvested seeds were analyzed for mineral, protein and fatty acid contents. The effect of the above treatment on seeds mineral was published in 2015. Results of the seed protein oil and fatty acids will be discussed in this presentation. Briefly, Mn, Cu and B treatments increased protein. Zn, Mo, Cu + CA, B + CA decreased the protein. Zn, Mo, CA, Cu+CA, Zn+CA, Mo+CA and B+CA increased the oil Mn, Cu decreased the oil. The Cu and B treatments increased the oleic acid (18:1) by 8.0 and 7.4%, respectively. Treatments, Mn, Mo, CA, and combo treatments Mn+CA, Cu+CA, Zn+CA, Mo+CA and B+CA all decreased oleic acid (18:1) by 0.6 to 14.4%. Cu, Zn, Mo, B, CA, Mn and in combination with CA increased linoleic acid by 1.3 to 6.5 %. Our goal is to identify the elements that would make desirable changes in the composition of the soybean seeds.
North-West University, South Africa
Title: Calmodulin dependent protein kinase (CaMK) II activation by exercise regulates lipid metabolism in rat skeletal muscle
Time : 12:30-13:00
Sandile Fuku has completed his Doctoral studies in Biomedical Technology from the Central University of Technology and is currently a Post-doctoral fellow in the Department of Biochemistry at North-West University. Currently, his research is on epigenetic regulation in metabolic syndromes, particularly focusing on Diabetics and Cancer. He has published work in Cancer Treatment, Diabetes and Phytochemistry.
Background: Activation of calmodulin dependent protein kinase (CaMK) II by exercise has plethora of benefits in metabolism and health. Regulation of lipid metabolism is very significant to alleviate type 2 diabetes and obesity. The role of CaMKII in the regulation of genes that are involved in lipid metabolism has not been studied yet, which became the focus of this study.
Methods: 5 to 6 weeks old male Wistar rats were used in this study. Western blot was performed to assess the protein expression of carnitine palmitoyltransferase (CPT)-1 and acetyl-CoA carboxylase (ACC)-1. Cpt-1 and Acc-1 gene expressions were assessed using quantitative real time PCR (qPCR).
Results: The results indicate that exercise-induced CaMKII activation increases CPT-1 expression and decreases ACC-1 expression in rat skeletal muscle. Thus, confirming CaMKII activation by exercise and the resultant increase in lipid oxidation. Administration of KN93 (CaMKII inhibitor) reversed all exercise-induced changes.
Conclusions: This study demonstrated that CaMKII activation, by exercise, regulates lipid metabolism genes in rat skeletal muscle. Further, the increase in lipid oxidation and decrease in lipid synthesis are evidence of the regulatory role CaMKII in lipid metabolism. CaMKII is a potential target in designing novel therapeutic drugs in the management and treatment of type 2 diabetes and obesity.
University of Babylon, Iraq
Title: Biochemical study of enzymatic antioxidant for the impact of benzene pollution on Albino Swiss mice as a risk factor of acute leukemia
Time : 14:00-14:30
Mufeed Jalil Ewadh has his expertise in Biochemistry. He has participated in many international and local conferences and workshops which deal with improvement of biochemical research to increase people awareness about its role. He has participated in Post- doctorate course in Marburg University (Germany) in 2005, and has participated in Electrophoresis Workshop in japan for two weeks as well as in Biochemical Workshop in Leipzig (Germany), 2016. He has published more than 134 papers in different local and international journals.
Theoretical: Leukemia is cancer of blood-forming tissues which starts in the bone marrow; characterized by highly elevated abnormal proliferation and circulation of immature clonal hematopoietic cells, hence leukemia is commonly referred to hematological neoplasms. There are two main categories of leukemia etiology: inherited and acquired. Several factors have been implicated in the causation of acquired Leukemia such as pollution and exposure to chemicals such as benzene.
Aim of the Study: The present study aimed to evaluate activity and efficiency of enzymatic antioxidant system - representative with GSH-Px and GST in case of acquired leukemia injury.
Methodology: The present study included (30) albino male mice divided into two groups with count (15) mice for each. Group (A) was the control group while group (B) was induced leukemia mice by subcutaneous injection of (300 mg/kg BW benzene). Shimadzu UV-Vis spectrophotometer has been used for estimation of GSH-Px and GST activities using special kits.
Conclusion: This study concluded that leukemia leads to imbalance oxidant-antioxidant system causing high oxidative stress and ROS production which in turn causes nonspecific oxidative damage to biomolecules in myeloid cells resulting in the development/ increase in leukemia.
University Children’s Hospital Bochum, Germany
Title: The l-arginine/NO pathway, homoarginine, and nitrite-dependent renal carbonic anhydrase activity in young people with type 1 diabetes mellitus
Time : 14:30-15:00
Christina Carmann is a Doctoral candidate in the Department of Neuropediatrics of the University Children’s Hospital in Bochum, Germany. The results of her dissertation were published in the Journal of Amino Acids in July 2015 entitled “The l-arginine/NO pathway, homoarginine, and nitrite-dependent renal carbonic anhydrase activity in young people with type 1 diabetes mellitus”. Her main focus of the study is the L-Arginin/NO-pathway in different pediatric diseases. She is working in the University Hospital of Berne, Switzerland, Department of Pediatric Surgery.
High circulating levels of asymmetric dimethylarginine (ADMA) and low circulating levels of homoarginine (hArg) are known cardiovascular risk factors in adults. While in adults with type 1 diabetes mellitus (T1DM) circulating ADMA is significantly elevated, in children and adolescents the reported ADMA data are contradictory. In 102 children with T1DM and 95 healthy controls (HC) serving as controls, we investigated the l-arginine (Arg)/nitric oxide (NO) pathway. Children with T1DM were divided into two groups, i.e., in children with newly diagnosed diabetes mellitus [T1DM-ND; n=10; age, 8.8 (4.4–11.2) years; HbA1c, 13 (8.9–13.9) %] and in those with long-term treatment [T1DM-T; n=92; age, 12.5 (10.5–15.4) years; HbA1c, 8.0 (7.2–8.6) %]. Amino acids and NO metabolites of the Arg/NO pathway and creatinine were measured by GC–MS or GC–MS/MS. There was a significant difference between T1DM-T and HC with regard to plasma nitrite [0.53 (0.48–0.61) vs. 2.05 (0.86–2.36) μM, P<0.0001] as well as to urinary nitrite [0.09 (0.06–0.17) vs. 0.22 (0.13– 0.37) μmol/mmol creatinine, P<0.0001]. Plasma, but not urinary nitrite, differed between T1DM-ND and HC [0.55 (0.50–0.66) vs. 2.05 (0.86–2.36) μM, P<0.0001]. The urinary nitrate-to-nitrite molar ratio (UNOXR), a measure of nitrite-dependent renal carbonic anhydrase (CA) activity, was higher in T1DM-T [1173 (738–1481), P<0.0001] and T1DM-ND [1341 (1117–1615), P=0.0007] compared to HC [540 (324–962)], but did not differ between T1DM-T and T1DM-ND (P=0.272). The lower nitrite excretion in the children with T1DM may indicate enhanced renal CA-dependent nitrite reabsorption compared with healthy children. The Arg/NO pathway is altered in T1DM in childhood and adolescence, yet the role and the importance of hArg and ADMA in T1DM remain to be elucidated.
Damanhour University, Egypt
Title: Effect of the polycyclic aromatic hydrocarbons exposure on sperm DNA in idiopathic male infertility
Time : 15:00-15:30
EL-Hassan Mokhamer has his expertise in Molecular and biochemical changes due to environmental pollution and passion in improving the health and his academic activities is teaching of cell and Molecular biology for Pharos pharmacy school students In Alexandria, Egypt. Also teaching Molecular Biology for zoology Department students, Faculty of Science, Damanhour University.
There is an increasing awareness of the potential role of genetic and environmental factors in idiopathic male infertility. However, there is little compelling evidence to date to suggest that the risk of idiopathic male infertility among the general population is influenced by exposure to certain chemicals. Thus the first objective of the present study is to assess the occurrence and distribution of PAHs in mussels of Alex Coast, to identify the origin of PAHs in the Alex Coast, Secondary, investigate the possible association between exposure to PAHs and male idiopathic infertility through; Estimation of urinary metabolites of PAHs, malonaldehyde (MDA), GSH, GST, testosterone, FSH, prolactin, Semen analysis and sperm chromatin dispersion test (Halo sperm). The present results of the study revealed that there were high concentration of many PAHs detected in the tissues of two species of mussels collected from Alex Coast which may supposed to be at big risk for human health. Also, The present results revealed that there was a high level of urinary 1-hydroxy pyrene, 1-hydroxy naphthalene, 2-hydroxy naphthalene in the urine of detected infertile group.In the current study, a high significant increase in the level of MDA in the sera of detected idiopathic infertile group was observed with a significant decrease in glutathione content. Where, the compounds resulting from the oxidation of PAHs have the ability to enter redox cycles, which increased the formation of reactive oxygen species (ROS) and thus caused sperm DNA damage. The data provide strong evidence that semen samples containing a statistical threshold of 30% sperm DNA fragmentation have a reduced level of pregnancy success. The results of the present study elucidated that there were DNA fragmentation from 32% - 40% in the sperm of some idiopathic infertility subjects.
Lead City University, Nigeria
Title: Anti-diabetic potential of Sapium ellipticum (Hochst) pax leaf extract in streptozotocin-induced diabetic wistar rats
Time : 15:30-16:00
Ighodaro O M is a Lecturer in the Department of Biochemistry, Lead City University, with research interest in Biochemical Toxicology and Medicinal Plant Research. His research drive is primarily to curtail mortality and health complications that result from herbal toxicity and wrong use of herbs in treatments of ailments/diseases in Africa. He has investigated a number of plants traditionally acclaimed to possess medicinal properties in a bid to appropriately advise and orientate individuals and communities on their proper usage. His ultimate goal is to possibly isolate new drug materials with relatively better therapeutic characteristics and less or allowable side chemistry from natural sources such as plants.
Ethnobotanical survey associates Sapium ellipticum (SE) with antidiabetic usage among other medicinal functions in different parts of Africa. In view of this, the effects of the plant leaf extract on fasting blood glucose (FBG), glycogen and insulin levels were investigated. SE was evaluated in streptozotocin-induced diabetic adult male Wistar rats, at 400 and 800 mg kg-1 of body weight (BW), against metformin (12 mg kg-1 BW). Treatments were done orally (p.o), twice daily at 8 h interval for 21 days. SE significantly reduced FBGL by 46.5 and 44.4 % (400 and 800 mg dosage, respectively) compared to initial diabetic values. However, the effects were significantly lower than the 72.6 % glucose reduction produced by metformin. Hepatic and skeletal muscle glycogens were observed to increase (27.06 and 12.55 % respectively) in SE-treated rats (800 mg dosage) compared to their corresponding values in diabetic control animals. Plasma and pancreatic insulin contents were also improved (31.77 and 52.34%, respectively) by SE administration. The histopathological examination of the pancreas indicates beta cells regeneration in the treated animals, particularly in diabetic rats treated with 800 mg dosage of the extract compared to the diabetic control animals and metformin group. H PLC-MS analysis of SE active fractions revealed the presence of amentoflavone, lupeol and luteolin-7-O-glucoside. The outcome of this study provides scientific basis in support of the medicinal relevance of SE and lend credence to its utilization in folk medicine for the treatment of diabetes and other oxidative stress-related ailments.
King Saud University, Saudi Arabia
Time : 16:20-16:50
Mohammed S Al-Khalifa completed his PhD from the Swansea University, Wales, UK in 1977 and ever since then is the faculty in the Zoology Department, King Saud University, Riyadh Saudi Arabia and actively involved in teaching graduate and post graduate students and supervised dozens of PhD and Master’s dissertation leading to the award of the degree of Doctor of Philosophy. He has published over 70 research articles on the different topics concerning medically important insects, ticks, mites and their physiology.
Investigations have been made on the anti-inflammatory effect of Samsum Ant Venom (SAV) in chemically-induced toxicity in male rats. Observations showed that CCl4 significantly raised the oxidative stress level in CCl4-treated rats. Significant rise of malondialdehyde (MDA) with a significant suppression of glutathione were detected in both hepatic and splenic tissues. Also increase in the total white blood cell counts with significantly high neutrophil percentage was observed in treated rats. In addition, CCl4 significantly increased the cholesterol, triglycerides, creatinine, urea and ALT, and decrease the total protein level. Treatment with 100 μg/kg dose of SAV was found to significantly restore all these studied parameters. It was found to significantly restore the glutathione level and thus decrease the oxidative stress effects. Furthermore, it successfully restored the biochemical parameters, namely, urea, creatinine, cholesterol and total protein to the normal levels. The positive correlation between SAV and the animal model health was clearly observed in the restoration of the normal architecture of both hepatic and splenic tissues with normal distribution of the immune cells in the white pulp of spleen treated with CCl4.
Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russia
Time : 16:50-17:20
Natalia V Povarova has got her Master’s degree in Biochemistry ("Study on interchangeability of picornaviral security proteins") at Lomonosov Moscow State University, Russia in 2011. Currently, she is doing her PhD degree in the Biophotonics Lab of Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry (Moscow, Russia) on Silicatein Biochemistry and its applications.
Silicateins are the major spicule-forming enzymes of marine sponges. They are members of the cathepsin family of cysteine proteases but are distinguished from cathepsins by Cys to Ser substitution in the catalytic triad (Cys-His-Asn). Silicateins catalyze formation of amorphous SiO2 from the silica precursors at physiological conditions. It is a promising biotechnological tool to obtain bio-silica covers and particles. The most popular silica precursor for silicatein is tetraethyl orthosilicate (TEOS). However, this compound is poorly soluble in water, and thus it is not readily available for the protein. We recently introduced a new substrate, tetrakis-(2-hydroxyethyl)-orthosilicate (THEOS), which is soluble in water, but its hydrolysis rate is very high, leading to a high level of spontaneous polymerization. In addition, TEOS, THEOS and its hydrolysis products are highly toxic for cells. Here we report a novel silica precursor for silicateins tetrakis (glycerol) orthosilicate (TGS). It is soluble in water and has a low spontaneous polymerization rate. Also, we found that TGS is much less toxic for mammalian cultured cells compared to TEOS and THEOS. We have tested TGS as a substrate for silicatein A1 from the marine sponge Latrunculia oparinae. It exhibited high activity with TGS and effectively formed silic particles. Water-solubility of TGS and appropriate activity level enabled us to measure for the first time silicatein’s biochemical propertiesactivity dependence on pH, temperature and precursor concentration. We also constructed several mutants of the silicatein A1 presumed active site triad (Ser25-His163-Asn187). Surprisingly, even Ala mutants (S25A, H163A, or N187A) retained a high activity in polymerization of TGS and THEOS, but not TEOS. These data call for reevaluation of the suggested earlier mechanisms of action and biochemical properties of silicateins.
- Video Presentation
Ajinomoto Co., Japan
Time : 17:20-17:30
San Gabriel A has expertise on Taste Physiology and especially, on G-protein-coupled receptors of taste receptors and gastric cells. She has also work on the analysis of non-protein free amino acids (FAA) in breast milk and is interested on understanding how dietary proteins from the mother influences on the FAA profile of her milk. Currently, she is working as a Science Communicator in the Global Communications department of Ajinomoto Co., and is the Scientific Affairs Representative of the NPO Umami Information Center.
Sensory receptors from eyes, nose, skin, and mouth notify the brain about the world that surrounds us; and all are essential to evaluate how appealing is a meal. Non-protein free amino acids (FAA) from foods seem to bind to various sensory receptors on the tongue, taste receptor cells, and in the stomach where different cells can detect FAA via the calcium sensing receptor (CaSR), the G-protein coupled receptor family C group 6 (GPRC6A), the taste receptor type 1 heterodimer (TAS1R1/TAS1R1), and the metabotropic glutamate receptors (mGluRs). Amino acid transporters in membranes of gastric cells can also regulate different cell functions. FAAs add characteristic flavors to certain foods with umami, bitter or sweet tastes, and in the stomach, before protein digestion takes place, they may interact with signaling receptors in various cells, from mucous, pepsinogen or acidproducing cells, to enteroendocrine cells that release gastrin, somatostatin, or ghrelin. All these hormones are crucial for the regulation of protein digestion in the stomach and the sensation of hunger after a meal. This suggests that there might be a link between taste, quality and intensity, and gastric function and appetite regulation. There is a need for a deeper understanding on the role of taste active compounds in the stomach because FAAs in foods may enhance the satiating effect of proteins via taste receptors besides the fact that appealing tastes with FAAs can promote the intake of more healthful ingredients such as vegetables or low fat low salt foods.
University of Malaya, Malaysia
Title: Electro-oxidation of Amino Acids in Parkia Speciose Using a Novel Ag2O/CNT/ND-based Electro-chemical Biosensor
Time : 17:30-17:40
She has completed her PhD from Universiti kebangsaan Malaysia and postdoctoral studies from Universsiti Putra Malaysia. She has published more than 45 papers in reputed journals. Her primary area of research has focused on fabrication anhydrous membrane in fuel cell application. She is working on several fields especially environment. Her specific interest is research on advanced materials and electrochemical reaction. She interested to work on biomedical field and fabrication biosensors for various disease and tissue engineering. Currently, she is working on biosensors and investigation of their performance via electrochemical reaction and impedance spectroscopy.
The increasing awareness of the importance of a healthy diet has led to the development of new, safe and healthy foods. Natural food-derived peptides with specific bioactivity have attracted a considerable interest among researchers. Parkia speciose (stink bean), a southeast Asian legume, is composed of chemically-medicinal compounds which exhibit biological activities. It is reported to be anticancer, antibacterial, antioxidant, antiangiogenic and demonstrates hem-agglutinating activity. The compositional analysis of amino acids in Parkia speciosa seeds have been reported through hydrolysis using alcalase enzyme. The cyclic voltammetry (CV) technique using biosensors has been well established with broad applications in quantitative and qualitative analyses. Our novel electro-chemical biosensor Ag2O/CNT/ND comprises silver-oxide, nano-diamond (ND) and carbon nanotube (CNT), fabricated on glassy carbon electrodes. The Ag2O/CNT/ND-based biosensor exhibited irreversible oxidation and reduction peaks at 0.85 V and -0.95 V in a phosphate buffer carrier (0.01 M, pH 6.8), respectively. At higher scan rates, the CVs were slightly shifted to the more negative potentials with a maximum peak at about −1.3 V. This suggested the formation of H2O2/H2S which was also confirmed by a reduction in pH value from 6.8 to 6.55. It was observed that the anodic and cathodic peak currents increased linearly with the square root of the scan rate (ν1/2) over the studied scan range of 0.01 – 0.1 V/s indicating a diffusion-controlled irreversible electro-chemical process. The developed biosensor displayed a very good electro-catalytic activity toward the oxidation of H2O2 and the release of H2S as a result of the reaction between the active sites and the Parkia speciose ingredient.