Search results for: alkaline single-cell gel electrophoresis

Authors: Syeda Hira, Muhammad Gulfraz

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Background: Liver diseases cause serious health issues. Plants contain active compounds that significantly help in the treatment of various diseases. Ficus carica is traditionally used for the treatment of liver diseases. The purpose of the present study was the isolation and identification of active components from F.carica leaves which are responsible for hepatoprotective activity. Methods: The study was designed to identify the most active hepatoprotective sub-fraction from ethyl acetate fraction of Ficus carica by in vitro study and evaluation of its in vivo hepatoprotective effect in animal models. Ethyl acetate fraction was subjected to column, and a total of eight sub-fractions were obtained. In vitro, the hepatoprotective effect of all sub-fractions was determined on HepG2 cell lines. Toxicity was induced by CCl₄ (Carbon tetrachloride), and silymarin was used as a positive control. On the basis of the results, the most active sub-fraction was subjected to LC-MS and FT-IR analysis for the identification of bioactive compounds. In vivo, the hepatoprotective effect was determined in mice. Toxicity was induced by CCl₄; at the end of the experiment, biochemical parameters such as ALT, AST, ALP, bilirubin, and total protein were estimated in serum. Histopathology of liver tissues was also done. Results: Sub-fraction FVI exhibited significant (P<0.05) hepatoprotective activity as compared to other sub-fractions, which was almost similar to the standard drug silymarin. Six known bioactive compounds were identified from this sub-fraction after LC-MS analysis. In vivo, the hepatoprotective activity of sub-fraction FVI was evaluated in CCl₄-induced toxicated mice. Administration of CCl₄ significantly increased level of ALT (Alanine transaminase), AST (Aspartate aminotransferase), ALP (Alkaline phosphatase), and bilirubin and decreased the total protein. Treatment with sub-fraction FVI significantly (p<0.05) reversed the level of these biomarkers toward normal at both doses of 25 mg/kg and 50 mg/kg. Conclusion: Our findings confirmed the hepatoprotective effect of ethyl acetate fraction of F.carica. It could be a good candidate for the development of a natural hepatoprotective drug; pre-clinical investigation on ethyl acetate fraction is recommended.

Keywords: Ficus carica, hepatoprotective, CCl₄, bioactive compounds, liver markers

Procedia PDF Downloads 43

Authors: Ugwuowo Leonard Chidi, Oparaji Chetachukwu Jecinta., Ogidi Chibuzor Agafenachukwu, Onuoha Rebecca Obianuju

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This experiment was conducted to investigate the growth performance, haematology and serum biochemistry indices of broiler birds fed diets containing Neem leaf meal. A total of 96 unsexed day-old broiler birds were allocated to four treatments of T1, T2, T3 and T4 and replicated three times with eight birds per replicate in a Completely Randomized Design. The treatments were diets containing 2.0, 4.0, 6.0 and 8.0% Neem leaf meal respectively. Growth performances, packed cell volume, red blood cell count, haemoglobin, white blood cell count, lymphocytes, mean corpuscular volume, mean corpuscular haemoglobin concentration, platelet count, aspartate amino transaminase, alanine amino transaminase, alkaline phosphate, cholesterol, albumin, globulin, urea, glucose, total protein and creatinine were evaluated. Results showed that there were no significant differences (P>0.05) in all the growth performance parameters among the treatments. The results of the experiment showed that there were significant differences (P<0.05) in all the heamatological and serum biochemistry parameters at finisher phases. Mean corpuscular volume, white blood cell count, lymphocytes, red blood cell count, haemoglobin, platelet count, creatinine and triglyceride increased and were highest in treatment two while treatment four had the least values in mean corpuscular volume, urea, white blood cell, haemoglobin and triglyceride. This implies that the levels of inclusion of Neem leaf meal in this experiment did not affect the growth performance of the broiler chicks but the haematological and serum biochemistry indices were affected. Treatment two with a 4% inclusion level of Neem leaf meal has shown the capacity to replace vaccines and antibiotics in broilers due to the positive effects it had on both the haematological and serum biochemistry.

Keywords: leaf meal, broiler, Aziradicta indica, serum biochemistry, haematology

Procedia PDF Downloads 53

Authors: Parminder Nain, Jaspreet kaur, Vipin Saini, Sunil Sharma

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Jasminum humile commonly known as yellow Jasmine or Pili chameli, is a medicinal plant used in Ayurveda for treating various diseases, one of which is diabetes mellitus. The current study aimed to establish the antidiabetic and antioxidant properties of aqueous extract of Jasminum humile leaves (AEJHL) in nicotinamide/streptozotocin induced type 2 diabetic rats. Phytochemical screening, HPLC analysis, and acute toxicity study of AEJHL were carried out. Male albino wistar rats (n=42) were divided into seven equal groups. Rats with moderate diabetes having hyperglycemia (blood glucose 250-400 mg/dl) were taken for the experiment. Various concentrations of aqueous extract of Jasminum humile leaves (50, 100, 200 and 300 mg/kg, p.o.), and glibenclamide (1mg/kg, p.o.) were orally administered to diabetic rats for 45 days. The effect of AEJHL on blood glucose, plasma insulin and biochemical parameters such as hemoglobin, total protein, serum creatinine, serum urea, alkaline phosphate, Glutamic-oxalacetic transaminase (SGOT) and glutamic-pyruvic transaminase (SGPT), as well as total cholesterol, triglycerides, and high-density lipoprotein (HDL) were also studied. The antioxidant effect of AEJHL was determined by analyzing hepatic and renal antioxidant markers, like superoxide dismutase (SOD), catalase (CAT), reduced Glutathione (GSH), Glutathione peroxidase (GPx), and lipid peroxidation (LPO) in diabetic rats. After 45-days oral administration of aqueous extract of Jasminum humile leaves significantly (p<0.05) reduced blood sugar and increase plasma insulin level and also reverse all above biochemical parameters and antioxidant enzyme level at dose dependent manner. These findings provide in vivo evidence that the aqueous extract of Jasminum humile leaves possess significant antidiabetic and antioxidant potential in nicotinamide/streptozotocin-induced type-2 diabetes mellitus in rats.

Keywords: antidiabetic, antioxidant, jasminum humile, nicotinamide/streptozotocin, type-2 diabetic

Procedia PDF Downloads 183

Authors: Hanaa M. M. El-Khayat, Hanan S. Gaber, Hoda Abdel-Hamid, Kadria M. A. Mahmoud, Hoda M. A. Abu Taleb

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This work aimed to examine Oreochromis niloticus fish from Lake Manzala in Port Said, Dakahlya and Damietta governorates, Egypt, as a bio-indicator for the lake water pollution through recording alterations in their hematological, physiological, and histopathological parameters. All fish samples showed a significant increase in levels of alkaline phosphatase (ALP), creatinine and glutathione-S-transferase (GST); only Dakahlya samples showed a significant increase (p<0.01) in aspartate aminotransferase (AST) level and most Dakahlya and Damietta samples showed reversed albumin and globulin ratio and a significant increase in γ-glutamyltransferase (GGT) level. Port-Said and Damietta samples showed a significant decrease of hemoglobin (Hb) while Dakahlya samples showed a significant decrease in white blood cell (WBC) count. Histopathological investigation for different fish organs showed that Port-Said and Dakahlya samples were more altered than Damietta. The muscle and gill followed by intestine were the most affected organs. The muscle sections showed severe edema, neoplasia, necrotic change, fat vacuoles and splitting of muscle fiber. The gill sections showed dilated blood vessels of the filaments, curling of gill lamellae, severe hyperplasia, edema and blood vessels congestion of filaments. The intestine sections revealed degeneration, atrophy, dilation in blood vessels and necrotic changes in sub-mucosa and mucosa with edema in between. The recorded significant alterations, in most of the physiological and histological parameters in O. niloticus samples from Lake Manzala, were alarming for water pollution impacts on lake fish community, which constitutes the main diet and the main source of income for the people inhabiting these areas, and were threatening their public health and economy. Also, results evaluate the use of O. niloticus fish as important bio-indicator for their habitat stressors.

Keywords: Lake Manzala, Oreochromis niloticus fish, water pollution, physiological, hematological and histopathological parameters

Procedia PDF Downloads 291

Authors: Violina Angelova, Galina Pevicharova

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A comparative research had been carried out to allow us to determine the quantities and the centers of accumulation of Pb, Cu, Zn and Cd in the vegetative and reproductive organs of the sweet potatoes and to ascertain the possibilities for growing them on soils, polluted with heavy metals. The experiments were performed on agricultural fields contaminated by the (1) Non-Ferrous-Metal Works near Plovdiv, (2) Lead and Zinc Complex near Kardjali and (3) a copper smelter near Pirdop, Bulgaria. The soils used in this experiment were characterized by acid, neutral and slightly alkaline reaction, loamy texture and a moderate content of organic matter. The total content of Zn, Pb, and Cd was high and exceeded the limit value in agriculture soils. Sweet potatoes were in a 2-year rotation scheme on three blocks in the experimental field. On reaching commercial ripeness the sweet potatoes were gathered and the contents of heavy metals in their different parts – root, tuber (peel and core), leaves and stems, were determined after microwave mineralization. The quantitative measurements were carried out with inductively coupled plasma atomic emission spectroscopy. The contamination of the sweet potatoes was due mainly to the presence of heavy metals in the soil, which entered the plants through their root system, as well as by diffusion through the peel. Pb, Cu, Zn, and Cd were selectively accumulated in the underground parts of the sweet potatoes, and most of all in the root system and the peel. Heavy metals have an impact on the development and productivity of the sweet potatoes. The high anthropogenic contamination leads to an increased assimilation of heavy metals which reduces the yield and the quality of the production of sweet potatoes, as well as leads to decrease of the absolute dry substance and the quantity of sugars in sweet potatoes. Sweet potatoes could be grown on soils, which are light to medium polluted with lead, zinc, and cadmium, as they do not accumulate these elements. On heavily polluted soils, however, (Pb – 1504 mg/kg, Zn – 3322 mg/kg, Cd – 47 mg/kg) the growing of sweet potatoes is not allowed, as the accumulation of Pb and Cd in the core of the potatoes exceeds the Maximum Acceptable Concentration. Acknowledgment: The authors gratefully acknowledge the financial support by the Bulgarian National Science Fund (Project DFNI DH04/9).

Keywords: heavy metals, polluted soils, sweet potatoes, uptake

Procedia PDF Downloads 187

Authors: Esam Jassim

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Industries using conventional fossil fuels have an interest in better understanding the mechanism of particulate formation during combustion since such is responsible for emission of undesired inorganic elements that directly impact the atmospheric pollution level. Fine and ultrafine particulates have tendency to escape the flue gas cleaning devices to the atmosphere. They also preferentially collect on surfaces in power systems resulting in ascending in corrosion inclination, descending in the heat transfer thermal unit, and severe impact on human health. This adverseness manifests particularly in the regions of world where coal is the dominated source of energy for consumption. This study highlights the behavior of calcium transformation as mineral grains verses organically associated inorganic components during pulverized coal combustion. The influence of existing type of calcium on the coarse, fine and ultrafine mode formation mechanisms is also presented. The impact of two sub-bituminous coals on particle size and calcium composition evolution during combustion is to be assessed. Three mixed blends named Blends 1, 2, and 3 are selected according to the ration of coal A to coal B by weight. Calcium percentage in original coal increases as going from Blend 1 to 3. A mathematical model and a new approach of describing constituent distribution are proposed. Analysis of experiments of calcium distribution in ash is also modeled using Poisson distribution. A novel parameter, called elemental index λ, is introduced as a measuring factor of element distribution. Results show that calcium in ash that originally in coal as mineral grains has index of 17, whereas organically associated calcium transformed to fly ash shown to be best described when elemental index λ is 7. As an alkaline-earth element, calcium is considered the fundamental element responsible for boiler deficiency since it is the major player in the mechanism of ash slagging process. The mechanism of particle size distribution and mineral species of ash particles are presented using CCSEM and size-segregated ash characteristics. Conclusions are drawn from the analysis of pulverized coal ash generated from a utility-scale boiler.

Keywords: coal combustion, inorganic element, calcium evolution, fluid dynamics

Procedia PDF Downloads 313

Authors: Maria Giovanna Lupo, Marina Camera, Marcello Rattazzi, Nicola Ferri

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A complex interplay among chronic kidney disease, lipid metabolism and aortic calcification has been recognized starting from results of many clinical and experimental studies. Here we investigated the influence of kidney function on PCSK9 levels, both in uremic rats and in clinical observation study, and its potential direct action on cultured smooth muscle cells (SMCs) calcification. In a cohort of 594 subjects enrolled in a single centre, observational, cross-sectional and longitudinal study, a negative association between GFR and plasma PCSK9 was found. Atherosclerotic cardiovascular disease (ASCVD), as co-morbidity, further increased PCSK9 plasma levels. Diet-induced uremic condition in rats, induced aortic calcification and increased total cholesterol and PCSK9 levels in plasma, livers and kidneys. Immunohistochemical analysis confirmed PCSK9 expression in aortic SMCs. SMCs overexpressing PCSK9 (SMCsPCSK9), cultured for 7-days in a pro-calcification environment (2.0mM or 2.4mM inorganic phosphate, Pi) showed a significantly higher extracellular calcium (Ca2+) deposition compared to mocked SMCs. Under the same experimental conditions, the addition of exogenous recombinant PCSK9 did not increase the extracellular calcification of SMCs. By flow cytometry analysis we showed that SMCsPCSK9, in response to 2.4mM Pi, released higher number of extracellular vesicles (EVs) positive for three tetraspanin molecules, such as CD63, CD9, and CD81. EVs derived from SMCsPCSK9 tended to be more enriched in calcium and alkaline phosphatase (ALPL), compared to EVs from mocks SMCs. In conclusion, our study reveals a direct role of PCSK9 on vascular calcification induced by higher inorganic phosphate levels associated to CKD condition. This effect appears to be mediated by a positive effect of endogenous PCSK9 on the release of EVs containing Ca2+ and ALP, which facilitate the deposition inorganic calcium phosphate crystals.

Keywords: PCSK9, calcification, extracellular vesicles, chronic kidney disease

Procedia PDF Downloads 98

Authors: Shih-Ping Liu, Cheng-Hsuan Chang, Yu-Chuen Huang, Shih-Yin Chen, Woei-Cherng Shyu

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Embryonic stem cells (ES) and induced pluripotnet stem cells (iPS) are both pluripotent stem cells. For mouse stem cells culture technology, leukemia inhibitory factor (LIF) was used to maintain the pluripotency of stem cells in vitro. However, LIF is an expensive reagent. The goal of this study was to find out a pure compound extracted from Chinese herbal medicine that could maintain stem cells pluripotency to replace LIF and improve the iPS generation efficiency. From 20 candidates traditional Chinese medicine we found that Cordycepsmilitaris triggered the up-regulation of stem cells activating genes (Oct4 and Sox2) expression levels in MEF cells. Cordycepin, a major active component of Cordycepsmilitaris, also could up-regulate Oct4 and Sox2 gene expression. Furthermore, we used ES and iPS cells and treated them with different concentrations of Cordycepin (replaced LIF in the culture medium) to test whether it was useful to maintain the pluripotency. The results showed higher expression levels of several stem cells markers in 10 μM Cordycepin-treated ES and iPS cells compared to controls that did not contain LIF, including alkaline phosphatase, SSEA1, and Nanog. Embryonic body formation and differentiation confirmed that 10 μM Cordycepin-containing medium was capable to maintain stem cells pluripotency after four times passages. For mechanism analysis, microarray analysis indicated extracellular matrix and Jak/Stat signaling pathway as the top two deregulated pathways. In ECM pathway, we determined that the integrin αVβ5 expression levels and phosphorylated Src levels increased after Cordycepin treatment. In addition, the phosphorylated Jak2 and phosphorylated Sat3 protein levels were increased after Cordycepin treatment and suppressed with the Jak2 inhibitor, AG490. The expression of cytokines associated with Jak2/Stat3 signaling pathway were also up-regulated by Q-PCR and ELISA assay. Lastly, we used Oct4-GFP MEF cells to test iPS generation efficiency following Cordycepin treatment. We observed that 10 Μm Cordycepin significantly increased the iPS generation efficiency in day 21. In conclusion, we demonstrated Cordycepin could maintain the pluripotency of stem cells through both of ECM and Jak2/Stat3 signaling pathway and improved iPS generation efficiency.

Keywords: cordycepin, iPS cells, Jak2/Stat3 signaling pathway, molecular biology

Procedia PDF Downloads 420

Authors: I. A. Yola

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Impact of poultry dropping, cow dung and rumen content on utilization and proximate composition of Oreochromis niliticus, Clarias gariepinus and Cyprinus capio in a polyculture system were studied. The research was conducted over a period of 52 weeks. Poultry droppings (PD), cow dung (CD) and rumen content (RC) were applied at three levels 30g,60g and 120g/m2/week, 25g,50g and 100g/m2/week and 22g, 44g and 88g/m2/week treatment, respectively. The control only conventional feed with 40% CP without manure application was used. Physicochemical and biological properties measured were higher in manure pond than control. The difference was statistically significant (P < 0.05) between and within treatments with exception of temperature with a combined mean of 27.900C. The water was consistently alkaline with mean values for pH of 6.61, transparency 22.6cm, conductivity 35.00µhos/cm, dissolved oxygen 4.6 mg/l, biological oxygen demand 2.8mg/l, nitrate and phosphates 0.9mg/l and 0.35mg/l, respectively. The three fish species increase in weight with increased manure rate, with a higher value in PD treatment on C. capio record 340g, O. niloticus weighed 310g and C. gariepinus 280g over the experimental period. Fishes fed supplementary diet (control) grew bigger with highest value on C. capio (685g) O. niloticus (620g) and then C. gariepinus (526g). The differences were statistically significant (P < 0.05). The result of whole body proximate analysis indicated that various manures and rates had an irregular pattern on the protein and ash gain per 100g of fish body weight gain. The combined means for whole fish carcass protein, lipids, moisture, ash and gross energy were 11.84, 2.43, 74.63, 3.00 and 109.9 respectively. The notable exceptions were significant (p < 0.05) increases in body fat and gross energy gains in all fish species accompanied by decreases in percentages of moisture as manure rates increased. Survival percentage decreases from 80% to 70%. It is recommended to use poultry dropping as manure/feeds at the rate of 120kg/ha/week for good performances in polyculture.

Keywords: organic manure, Nile tilapia, African mud fish, common carp, proximate composition

Procedia PDF Downloads 531

Authors: Sachil Kumar, Anoop Kumar Verma, Wahid Ali, Uma Shankar Singh

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Globally approximately 55.3 million people die each year. In the India there were 95 lakh annual deaths in 2013. The number of deaths resulted from homicides, suicides and unintentional injuries in the same period was about 5.7 lakh. The ever-increasing crime rate necessitated the development of methods for determining time since death. An erroneous time of death window can lead investigators down the wrong path or possibly focus a case on an innocent suspect. In this regard a research was carried out by analyzing the temperature dependent degradation of a Cardiac Troponin-T protein (cTnT) in the myocardium postmortem as a marker for time since death. Cardiac tissue samples were collected from (n=6) medico-legal autopsies, (in the Department of Forensic Medicine and Toxicology, King George’s Medical University, Lucknow India) after informed consent from the relatives and studied post-mortem degradation by incubation of the cardiac tissue at room temperature (20±2 OC), 12 0C, 25 0C and 37 0C for different time periods ((~5, 26, 50, 84, 132, 157, 180, 205, and 230 hours). The cases included were the subjects of road traffic accidents (RTA) without any prior history of disease who died in the hospital and their exact time of death was known. The analysis involved extraction of the protein, separation by denaturing gel electrophoresis (SDS-PAGE) and visualization by Western blot using cTnT specific monoclonal antibodies. The area of the bands within a lane was quantified by scanning and digitizing the image using Gel Doc. The data shows a distinct temporal profile corresponding to the degradation of cTnT by proteases found in cardiac muscle. The disappearance of intact cTnT and the appearance of lower molecular weight bands are easily observed. Western blot data clearly showed the intact protein at 42 kDa, two major (27 kDa, 10kDa) fragments, two additional minor fragments (32 kDa) and formation of low molecular weight fragments as time increases. At 12 0C the intensity of band (intact cTnT) decreased steadily as compared to RT, 25 0C and 37 0C. Overall, both PMI and temperature had a statistically significant effect where the greatest amount of protein breakdown was observed within the first 38 h and at the highest temperature, 37 0C. The combination of high temperature (37 0C) and long Postmortem interval (105.15 hrs) had the most drastic effect on the breakdown of cTnT. If the percent intact cTnT is calculated from the total area integrated within a Western blot lane, then the percent intact cTnT shows a pseudo-first order relationship when plotted against the log of the time postmortem. These plots show a good coefficient of correlation of r = 0.95 (p=0.003) for the regression of the human heart at different temperature conditions. The data presented demonstrates that this technique can provide an extended time range during which Postmortem interval can be more accurately estimated.

Keywords: degradation, postmortem interval, proteolysis, temperature, troponin

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Authors: Boudjelal Amel

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Arisarum vulgare, a member of the Araceae family, is an herbaceous perennial widely distributed in the Mediterranean region. A. vulgare is recognized for its medicinal properties and holds significant traditional importance in Algeria for the treatment of various human ailments, including pain, infections, inflammation, digestive disorders, skin problems, eczema, cancer, wounds, burns and gynecological diseases. Despite its extensive traditional use, scientific exploration of A. vulgare remains limited. The study aims to investigate for the first time the therapeutic potential of A. vulgare ethanolic extract obtained by ultrasound-assisted extraction. The chemical composition of the extract was determined by LC-MS/MS analysis. For in vitro phytopharmacological evaluation, several assays, including DPPH, ABTS, FRAP and reducing power, were employed to evaluate the antioxidant activity. The antibacterial activity was assessed againt Escherichia coli, Salmonella typhimurium, Staphylococus aureus, Enterococcus feacium by disk diffusion and microdilution methods. The possible inhibitory activity of ethanolic extract was analyzed against the cholinesterases enzymes (AChE and BChE). The DNA protection activity of A. vulgare ethanolic extract was estimated using the agarose gel electrophoresis method. The capacities of the extract to protect plasmid DNA (pBR322) from the oxidizing effects of H2O2 and UV treatment were evaluated by their DNA-breaking forms. The in vivo wound healing potential of a traditional ointment containing 5% of A. vulgare ethanolic extract was also investigated. The LC-MS/MS profiling of the extract revealed the presence of various bioactive compounds, including naringenin, chlorogenic, vanillic, cafeic, coumaric acids, trans-cinnamic and trans ferrulic acids. The plant extract presented considerable antioxidant potential, being the most active for Reducing power (0,07326±0.001 mg/ml) and DPPH (0.14±0.004 mg/ml). The extract showed the highest inhibition zone diameter against Enterococcus feacium (36±0.1 mm). The ethanolic extract of A. vulgare suppressed the growth of Staphylococus aureus, Escherichia coli and Salmonella typhimurium according to the MIC values. The extract of the plant significantly inhibited both AChE and BChE enzymes. DNA protection activity of the A. vulgare extract was determined as 90.41% for form I and 51.92% for form II. The in vivo experiments showed that 5% ethanolic extract ointment accelerated the wound healing process. The topical application of the traditional formulation enhanced wound closure (95,36±0,6 %) and improved histological parameters in the treated group compared to the control groups. The promising biological properties of Arisarum vulgare revealed that the plant could be appraised as a potential origin of bioactive molecules having multifunctional medicinal uses.

Keywords: arisarum vulgare, LC-MS/MS, antioxidant activity, antimicrobial activity, cholinesterases enzymes inhibition, dna-damage activity, in vivo wound healing

Procedia PDF Downloads 38

Authors: Rabia Göçmen, Gülşah Kanbur, Sinan Sefa Parlat

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Aim of this study is to determine the effects of cellular insulin receptor stimulators on performance, plasma glucose, high density lipoprotein (HDL), low density lipoprotein (LDL), total cholesterol, triglyceride, triiodothyronine (T3) and thyroxine (T4) hormone levels, and incubation features in the breeding Japanese quails (Coturnix japonica). In the study, a total of 84 breeding quails was used, 6 weeks’ age, 24 are male and 60, female. Rations used in experiment are 2900 kcal/kg metabolic energy and 20% crude protein. Water pH is calibrated to 7.45. Ration and water were administered ad-libitum to the animals. As metformin source, metformin-HCl was used and as chrome resource, chromium picolinate was used. Trial groups were formed as control group (basal ration), metformin group (basal ration, added metformin at the level of feed of 20 mg/kg), and chromium picolinate (basal ration, added feed of 1500 ppb Cr) group. When regarded to the results of performance at the end of experiment, it is seen that live weight gain, feed consumption, egg weight, feed conversion ratio (Feed consumption/ egg weight), and egg production were affected at the significant level (p < 0.05). When the results are evaluated in terms of incubation features, hatchability and hatchability of fertile egg ratio were not affected from the treatments. Fertility ratio was significantly affected by metformin and chromium picolinate treatments and fertility rose at the significant level compared to control group (p < 0.05). According to results of experiment, plasma glucose level was not affected by metformin and chromium picolinate treatments. Plasma, total cholesterol, HDL, LDL, and triglyceride levels were significantly affected from insulin receptor stimulators added to ration (p < 0.05). Hormone level of Plasma T3 and T4 were also affected at the significant level from insulin receptor stimulators added to ration (p < 0.05).

Keywords: chromium picolinate, cholesterol, hormone, metformin, quail

Procedia PDF Downloads 203

Authors: Sairi Satari

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Introduction: Six-sigma is a metric that quantifies the performance of processes as a rate of Defects-Per-Million Opportunities. Sigma methodology can be applied in chemical pathology laboratory for evaluating process performance with evidence for process improvement in quality assurance program. In the laboratory, these methods have been used to improve the timeliness of troubleshooting, reduce the cost and frequency of quality control and minimize pre and post-analytical errors. Aim: The aim of this study is to evaluate the sigma values of the Cobas 8000 analyzer based on the minimum requirement of the specification. Methodology: Twenty-one analytes were chosen in this study. The analytes were alanine aminotransferase (ALT), albumin, alkaline phosphatase (ALP), Amylase, aspartate transaminase (AST), total bilirubin, calcium, chloride, cholesterol, HDL-cholesterol, creatinine, creatinine kinase, glucose, lactate dehydrogenase (LDH), magnesium, potassium, protein, sodium, triglyceride, uric acid and urea. Total error was obtained from Clinical Laboratory Improvement Amendments (CLIA). The Bias was calculated from end cycle report of Royal College of Pathologists of Australasia (RCPA) cycle from July to December 2016 and coefficient variation (CV) from six-month internal quality control (IQC). The sigma was calculated based on the formula :Sigma = (Total Error - Bias) / CV. The analytical performance was evaluated based on the sigma, sigma > 6 is world class, sigma > 5 is excellent, sigma > 4 is good and sigma < 4 is satisfactory and sigma < 3 is poor performance. Results: Based on the calculation, we found that, 96% are world class (ALT, albumin, ALP, amylase, AST, total bilirubin, cholesterol, HDL-cholesterol, creatinine, creatinine kinase, glucose, LDH, magnesium, potassium, triglyceride and uric acid. 14% are excellent (calcium, protein and urea), and 10% ( chloride and sodium) require more frequent IQC performed per day. Conclusion: Based on this study, we found that IQC should be performed frequently for only Chloride and Sodium to ensure accurate and reliable analysis for patient management.

Keywords: sigma matrics, analytical performance, total error, bias

Procedia PDF Downloads 154

Authors: Wighens Ngoie Ilunga, Pamela J. Welz, Olewaseun O. Oyekola, Daniel Ikhu-Omoregbe

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Currently, most sludge from the wastewater treatment plants of edible oil factories is disposed to landfills, but landfill sites are finite and potential sources of environmental pollution. Production of biodiesel from wastewater sludge can contribute to energy production and waste minimization. However, conventional biodiesel production is energy and waste intensive. Generally, biodiesel is produced from the transesterification reaction of oils with alcohol (i.e., Methanol, ethanol) in the presence of a catalyst. Homogeneously catalysed transesterification is the conventional approach for large-scale production of biodiesel as reaction times are relatively short. Nevertheless, homogenous catalysis presents several challenges such as high probability of soap. The current study aimed to reuse wastewater sludge from the edible oil industry as a novel feedstock for both monounsaturated fats and bioethanol for the production of biodiesel. Preliminary results have shown that the fatty acid profile of the oilseed wastewater sludge is favourable for biodiesel production with 48% (w/w) monounsaturated fats and that the residue left after the extraction of fats from the sludge contains sufficient fermentable sugars after steam explosion followed by an enzymatic hydrolysis for the successful production of bioethanol [29% (w/w)] using a commercial strain of Saccharomyces cerevisiae. A novel nano-magnetic catalyst was synthesised from mineral processing alkaline tailings, mainly containing dolomite originating from cupriferous ores using a modified sol-gel. The catalyst elemental chemical compositions and structural properties were characterised by X-ray diffraction (XRD), scanning electron microscopy (SEM), Fourier transform infra-red (FTIR) and the BET for the surface area with 14.3 m²/g and 34.1 nm average pore diameter. The mass magnetization of the nano-magnetic catalyst was 170 emu/g. Both the catalytic properties and reusability of the catalyst were investigated. A maximum biodiesel yield of 78% was obtained, which dropped to 52% after the fourth transesterification reaction cycle. The proposed approach has the potential to reduce material costs, energy consumption and water usage associated with conventional biodiesel production technologies. It may also mitigate the impact of conventional biodiesel production on food and land security, while simultaneously reducing waste.

Keywords: biodiesel, bioethanol, edible oil wastewater sludge, nano-magnetism

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Authors: Mahabub Alam, Mohammad Mahmudul Hassan, M. Hasanuzzaman, M. Ahasanul Hoque

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Animals are transported from one place to another for different purposes in Bangladesh. However, the potential effect of long-distance transport on cattle health has not frequently been studied. Therefore, this study was conducted to assess health conditions of cattle transported from a long distance to Chittagong in Bangladesh. A total of 100 adult cattle, regardless of breed and sex, were selected at Benapole live cattle market in Jessore between August and September 2015 for the study. Blood samples were taken from 50 randomly selected cattle at 0 hours before transportation, just after transportation, at 12-16 hours post-conclusion of transportation, and 24 hours after transportation. The external health conditions and injuries of the cattle were assessed by close inspection, and the trader was interviewed using the structured questionnaire. Images of cattle injuries were taken with a camera. The basic internal health of the cattle was evaluated using standard hemato-biochemical tests. Animals were fasted and remained standing within a small space allocation (8-10 sq feet/animal) in the vehicle during transportation. Animals were provided only with paddy straw and water prior to selling at the destination market. The overall frequency of cattle injuries varied significantly (26% before vs. 47% after transportation; p < 0.001). The frequency of different cattle injuries also significantly varied by types such as abrasion (11% vs. 21%; p < 0.05) and barbed wire injury (9% vs. 18%; p < 0.05). Single cattle injury differed significantly (21% vs. 36%; p < 0.001). Cattle health conditions varied significantly (nasal discharge: 15% vs. 28%; p < 0.05; diarrhea: 15% vs. 23%; p < 0.05 and severe dehydration: 8% vs. 20%; p < 0.001). The values of hemoglobin (Hb), total erythrocyte count (TEC), total leukocyte count (TLC), lymphocyte (L), neutrophil (N) and eosinophil (E) varied significantly (p ≤ 0.01) (Hb: 11.1mg/dl vs. 12.3mg/dl; TEC: 4.7 million/ml vs. 5.7million/ml; TLC: 6.2 thousand/ml vs. 7.3 thousand/ml; L: 61.7% vs. 58.1%; N: 29.7% vs. 32.8%; E: 3.8% vs. 4.7%). The values of serum total protein (TP), creatine kinase (CK), triglyceride (TG), calcium (Ca), phosphorus (P) and alkaline phosphatase (ALP) significantly differed (p ≤ 0.05) (TP: 6.8g/dl vs. 8.2g/dl; CK:574.9u/l vs. 1288u/l; TG: 104.7mg/dl vs. 127.7mg/dl; Ca: 11.3mg/dl vs. 13mg/dl; P: 7.3mg/dl vs. 7.6mg/dl; ALP: 303u/l vs. 363u/l). The identified status of external and internal health conditions of the cattle for trading purpose due to long-distance transportation in the present study indicates a high degree of transport stress and poor animal welfare.

Keywords: animal welfare, cattle, external and internal health conditions, transportation

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Authors: Hristina Šalipur, Jasmina Dostanić, Davor Lončarević, Matej Huš

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Photocatalytic water splitting/alcohol photoreforming has been used for the conversion of sunlight energy in the process of hydrogen production due to its sustainability, environmental safety, effectiveness and simplicity. Titanate nanotubes are frequently studied materials since they combine the properties of photo-active semiconductors with the properties of layered titanates, such as the ion-exchange ability. Platinum (Pt) doping into titanate structure has been considered an effective strategy in better separation efficiency of electron-hole pairs and lowering the overpotential for hydrogen production, which results in higher photocatalytic activity. In our work, Pt doped titanate catalysts were synthesized via simple alkaline hydrothermal treatment, incipient wetness impregnation method and temperature-programmed reduction. The structural, morphological and optical properties of the prepared catalysts were investigated using various characterization techniques such as X-ray diffraction (XRD), scanning electron microscopy (SEM), N2 physisorption, and diffuse reflectance spectroscopy (DRS). The activities of the prepared Pt-doped titanate photocatalysts were tested for hydrogen production via photocatalytic water splitting/alcohol photoreforming process under simulated solar light irradiation. Characterization of synthesized Pt doped titanate catalysts showed crystalline anatase phase, preserved nanotubular structure and high specific surface area. The result showed enhancement of activity in photocatalytic water splitting/alcohol photoreforming in the following order 2-butanol>1-butanol>tert-butanol, with obtained maximal hydrogen production rate of 7.5, 5.3 and 2 mmol g-1 h-1, respectively. Different possible factors influencing the hole scavenging ability, such as hole scavenger redox potential and diffusivity, adsorption and desorption rate of the hole scavenger on the surface and stability of the alcohol radical species generated via hole scavenging, were investigated. The theoretical evaluation using density functional theory (DFT) further elucidated the reaction kinetics and detailed mechanism of photocatalytic water splitting/alcohol photoreforming.

Keywords: hydrogen production, platinum, semiconductor, water splitting, density functional theory

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Authors: Xianghong Lu, Yuejun Wang, Chengshi Gan, Xin Qian

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The North Sulawesi Arc, located on the east of Indonesia and to the south of the Celebes Sea, is the north part of the K-shape of Sulawesi Island and has a complex tectonic history since the Cenozoic due to the convergence of three plates (Eurasia, India-Australia and Pacific plates). Published rock records contain less precise chronology, mostly using K-Ar dating, and rare geochemistry data, which limit the understanding of the regional tectonic setting. This study presents detailed zircon U-Pb geochronological and Hf-O isotope and whole-rock geochemical analyses for the Na-rich granites from the North Sulawesi Arc. Zircon U-Pb geochronological analyses of three representative samples yield weighted mean ages of 30.4 ± 0.4 Ma, 29.5 ± 0.2 Ma, and 27.3 ± 0.4 Ma, respectively, revealing the Oligocene magmatism in the North Sulawesi Arc. The samples have high Na₂O and low K₂O contents with high Na₂O/K₂O ratios, belonging to Low-K tholeiitic Na-rich granites. The Na-rich granites are characterized by high SiO₂ contents (75.05-79.38 wt.%) and low MgO contents (0.07-0.91 wt.%) and show arc-like trace elemental signatures. They have low (⁸⁷Sr/⁸⁶Sr)i ratios (0.7044-0.7046), high εNd(t) values (from +5.1 to +6.6), high zircon εHf(t) values (from +10.1 to +18.8) and low zircon δ18O values (3.65-5.02). They show an Indian-Ocean affinity of Pb isotopic compositions with ²⁰⁶Pb/²⁰⁴Pb ratio of 18.16-18.37, ²⁰⁷Pb/²⁰⁴Pb ratio of 15.56-15.62, and ²⁰⁸Pb/²⁰⁴Pb ratio of 38.20-38.66. These geochemical signatures suggest that the Oligocene Na-rich granites from the North Sulawesi Arc formed by partial melting of the juvenile oceanic crust with sediment-derived fluid-related metasomatism in a subducting setting and support an intra-oceanic arc origin. Combined with the published study, the emergence of extensive calc-alkaline felsic arc magmatism can be traced back to the Early Oligocene period, subsequent to the Eocene back-arc basalts (BAB) that share similarity with the Celebes Sea basement. Since the opening of the Celebes Sea started from the Eocene (42~47 Ma) and stopped by the Early Oligocene (~32 Ma), the geodynamical mechanism of the formation of the Na-rich granites from the North Sulawesi Arc during the Oligocene might relate to the subduction of the Indian Ocean.

Keywords: North Sulawesi Arc, oligocene, Na-rich granites, in-situ zircon Hf–O analysis, intra-oceanic origin

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Authors: Fama Jallow, Melody Neaves, Professor Mcgregor

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The quest for sustainable energy sources has driven significant interest in hydrogen production as a clean and efficient fuel. Alkaline water electrolysis (AWE) has emerged as a prominent method for generating hydrogen, necessitating the development of advanced electrode designs with improved performance characteristics. Additive manufacturing (AM) by laser powder bed fusion (LPBF) method presents an opportunity to tailor electrode microstructures and properties, enhancing their performance. This research proposes investigating the AM of electrodes with different lattice structures to optimize hydrogen production. The primary objective is to employ advanced modeling techniques to identify and select two optimal lattice structures for electrode fabrication. LPBF will be used to fabricate electrodes with precise control over lattice geometry, pore size, and distribution. The performance evaluation will encompass energy consumption and porosity analysis. AWE will assess energy efficiency, aiming to identify lattice structures with enhanced hydrogen production rates and reduced power requirements. Computed tomography (CT) scanning will analyze porosity to determine material integrity and mass transport characteristics. The research aims to bridge the gap between AM and hydrogen production by investigating lattice structures potential in electrode design. By systematically exploring lattice structures and their impact on performance, this study aims to provide valuable insights into the design and fabrication of highly efficient and cost-effective electrodes for AWE. The outcomes hold promise for advancing hydrogen production through AM. The research will have a significant impact on the development of sustainable energy sources. The findings from this study will help to improve the efficiency of AWE, making it a more viable option for hydrogen production. This could lead to a reduction in our reliance on fossil fuels, which would have a positive impact on the environment. The research is also likely to have a commercial impact. The findings could be used to develop new electrode designs that are more efficient and cost-effective. This could lead to the development of new hydrogen production technologies, which could have a significant impact on the energy market.

Keywords: hydrogen production, electrode, lattice structure, Africa

Procedia PDF Downloads 53

Authors: Masoud Sheidaei, Mohammad-Reza Kordasti, Fahimeh Koohdar

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Calotropis procera (Aiton) W.T.Aiton, (Apocynaceae), is an economically and medicinally important plant species which is an evergreen, perennial shrub growing in arid and semi-arid climates, and can tolerate very low annual rainfall (150 mm) and a dry season. The plant can also tolerate temperature ran off 20 to30°C and is not frost tolerant. This plant species prefers free-draining sandy soils but can grow also in alkaline and saline soils.It is found at a range of altitudes from exposed coastal sites to medium elevations up to 1300 m. Due to morpho-physiological adaptations of C. procera and its ability to tolerate various abiotic stresses. This taxa can compete with desirable pasture species and forms dense thickets that interfere with stock management, particularly mustering activities. Caloteropis procera grows only in southern part of Iran where in comprises a limited number of geographical populations. We used different population genetics and r landscape analysis to produce data on geographical populations of C. procera based on molecular genetic study using SCoT molecular markers. First, we used spatial principal components (sPCA), as it can analyze data in a reduced space and can be used for co-dominant markers as well as presence / absence data as is the case in SCoT molecular markers. This method also carries out Moran I and Mantel tests to reveal spatial autocorrelation and test for the occurrence of Isolation by distance (IBD). We also performed Random Forest analysis to identify the importance of spatial and geographical variables on genetic diversity. Moreover, we used both RDA (Redundency analysis), and LFMM (Latent factor mixed model), to identify the genetic loci significantly associated with geographical variables. A niche modellng analysis was carried our to predict present potential area for distribution of these plants and also the area present by the year 2050. The results obtained will be discussed in this paper.

Keywords: population genetics, landscape genetic, Calotreropis procera, niche modeling, SCoT markers

Procedia PDF Downloads 76

Authors: Erico R. Carmona, Lucas Hernandez-Saravia, Aliro Villacorta, Felipe Carevic

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Tailings and electronic waste (e-waste) are an important source of global contamination. Chile is one of Organisation for Economic Co-operation and Development (OECD) member countries that least recycled this kind of industrial waste, reaching only 3% of the total. Tailings and e-waste recycling offers a valuable tool to minimize the increasing accumulation of waste, supplement the scarcity of some raw materials and to obtain economic benefits through the commercialization of these. It should be noted that this type of industrial waste is an important source of valuable metals, such as copper, which allow generating new business and added value through its transformation into new materials with advanced physical and biological properties. In this sense, the development of nanotechnology has led to the creation of nanomaterials with multiple applications given their unique physicochemical properties. Among others, copper nanoparticles (CuNPs) have gained great interest due to their optical, catalytic, conductive properties, and particularly because of their broad-spectrum antimicrobial activity. There are different synthesis methods of copper nanoparticles; however, green synthesis is one of the most promising methodologies, since it is simple, low-cost, ecological, and generates stable nanoparticles, which makes it a promising methodology for scaling up. Currently, there are few initiatives that involve the development of methods for the recovery and transformation of copper from waste to produce nanoparticles with new properties and better technological benefits. Thus, the objective of this work is to show preliminary data about the develop a sustainable transformation process of tailings and e-waste that allows obtaining a copper-based nanotechnological product with potential antimicrobial applications. For this, samples of tailings and e-waste collected from Tarapacá and Antofagasta region of northern Chile were used to recover copper through efficient, ecological, and low-cost alkaline hydrometallurgical treatments, which to allow obtaining copper with a high degree of purity. On the other hand, the transformation process from recycled copper to a nanomaterial was carried out through a green synthesis approach by using vegetal organic residue extracts that allows obtaining CuNPs following methodologies previously reported by authors. Initial physical characterization with UV-Vis, FTIR, AFM, and TEM methodologies will be reported for CuNPs synthesized.

Keywords: nanomaterials, industrial waste, chile, recycling

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Authors: Nikhil John Kollannur, Dali Naidu Arnepalli

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Most of the surface related phenomena in the case of fine-grained soil are attributed to their unique surface charge properties and specific surface area. The temporal variations in soil behavior, to some extent, can be credited to the changes in these properties. Among the multitude of factors that affect the charge and surface area of clay minerals, the inherent system chemistry occupies the cardinal position. The impact is more profound when the chemistry change is manifested in terms of the system pH. pH plays a significant role by modifying the edge charges of clay minerals and facilitating mineral dissolution. Hence there is a need to address the variations in physical and charge properties of fine-grained soils treated over a range of acidic as well as alkaline conditions. In the present study, three soils (two soils commercially procured and one natural soil) exhibiting distinct mineralogical compositions are subjected to different pH environment over a range of 2 to 13. The soil-solutions prepared at a definite liquid to solid ratio are adjusted to the required pH value by adding measured quantities of 0.1M HCl/0.1M NaOH. The studies are conducted over a range of interaction time, varying from 1 to 96 hours. The treated soils are then analyzed for their physical properties in terms of specific surface area and particle size characteristics. Further, modifications in surface morphology are evaluated from scanning electron microscope (SEM) imaging. Changes in the surface charge properties are assessed in terms of zeta potential measurements. Studies show significant variations in total surface area, probably because of the dissolution of clay minerals. This observation is further substantiated by the morphological analysis with SEM imaging. The zeta potential measurements on soils indicate noticeable variation upon pH treatment, which is partially ascribed to the modifications in the pH-dependant edge charges and partially due to the clay mineral dissolution. The results provide valuable insight into the role of pH in a clay-electrolyte system upon surface related phenomena such as species adsorption, fabric modification etc.

Keywords: acid and alkali treatment, mineral dissolution , specific surface area, zeta potential

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Authors: Tiisetso Mpai, Sanjay K. Jaiswal, Felix D. Dakora

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The South African Cape fynbos biome is a global biodiversity hotspot. This biome contains a diversity of endemic shrub legumes, including Polhillia, Wiborgia, and Wiborgiella species, which are important for ecotourism as well as for improving soil fertility status. This is due to their proven N₂-fixing abilities when in association with compatible soil bacteria. In fact, Polhillia, Wiborgia, and Wiborgiella species have been reported to derive over 61% of their needed nitrogen through biological nitrogen fixation and to exhibit acid and alkaline phosphatase activity in their rhizospheres. Thus, their interactions with soil microbes may explain their survival mechanisms under the continued summer droughts and acidic, nutrient-poor soils in this region. However, information regarding their rhizosphere microbiome is still unavailable, yet it is important for Fynbos biodiversity management. Therefore, the aim of this study was to assess the microbial community structures associated with rhizosphere soils of Polhillia pallens, Polhillia brevicalyx, Wiborgia obcordata, Wiborgia sericea, and Wiborgiella sessilifolia growing at different locations of the South African Cape fynbos, during the wet and dry seasons. The hypothesis is that the microbial communities in these legume rhizospheres are the same type and are not affected by the growing season due to the restricted habitat of these wild fynbos legumes. To obtain the results, DNA was extracted from 0.5 g of each rhizosphere soil using PowerSoil™ DNA Isolation Kit, and sequences were obtained using the 16S rDNA Miseq Illumina technology. The results showed that in both seasons, bacteria were the most abundant microbial taxa in the rhizosphere soils of all five legume species, with Actinobacteria showing the highest number of sequences (about 30%). However, over 19.91% of the inhabitants in all five legume rhizospheres were unclassified. In terms of genera, Mycobacterium and Conexibacter were common in rhizosphere soils of all legumes in both seasons except for W. obcordata soils sampled during the dry season, which had Dehalogenimonas as the major inhabitant (6.08%). In conclusion, plant species and season were found to be the main drivers of microbial community structure in Cape fynbos, with the wet season being more dominant in shaping microbial diversity relative to the dry season. Wiborgia obcordata had a greater influence on microbial community structure than the other four legume species.

Keywords: 16S rDNA, Cape fynbos, endemic legumes, microbiome, rhizosphere

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Authors: Zatovska T. V., Nesterova N. V., Baranova G. V., Zagorodnya S. D.

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In recent years, biosensor technologies based on the phenomenon of surface plasmon resonance (SPR) are becoming increasingly used in biology and medicine. Their application facilitates exploration in real time progress of binding of biomolecules and identification of agents that specifically interact with biologically active substances immobilized on the biosensor surface (biochips). Special attention is paid to the use of Biosensor analysis in determining the antibody-antigen interaction in the diagnostics of diseases caused by viruses and bacteria. According to WHO, the diseases that are caused by the herpes simplex virus (HSV), take second place (15.8%) after influenza as a cause of death from viral infections. Current diagnostics of HSV infection include PCR and ELISA assays. The latter allows determination the degree of immune response to viral infection and respective stages of its progress. In this regard, the searches for new and available diagnostic methods are very important. This work was aimed to develop Biosensor chip for detection of specific antibodies to HSV-1 in the human blood serum. The proteins of HSV1 (strain US) were used as antigens. The viral particles were accumulated in cell culture MDBK and purified by differential centrifugation in cesium chloride density gradient. Analysis of the HSV1 proteins was performed by polyacrylamide gel electrophoresis and ELISA. The protein concentration was measured using De Novix DS-11 spectrophotometer. The device for detection of antigen-antibody interactions was an optoelectronic two-channel spectrometer ‘Plasmon-6’, using the SPR phenomenon in the Krechman optical configuration. It was developed at the Lashkarev Institute of Semiconductor Physics of NASU. The used carrier was a glass plate covered with 45 nm gold film. Screening of human blood serums was performed using the test system ‘HSV-1 IgG ELISA’ (GenWay, USA). Development of Biosensor chip included optimization of conditions of viral antigen sorption and analysis steps. For immobilization of viral proteins 0.2% solution of Dextran 17, 200 (Sigma, USA) was used. Sorption of antigen took place at 4-8°C within 18-24 hours. After washing of chip, three times with citrate buffer (pH 5,0) 1% solution of BSA was applied to block the sites not occupied by viral antigen. It was found direct dependence between the amount of immobilized HSV1 antigen and SPR response. Using obtained biochips, panels of 25 positive and 10 negative for the content of antibodies to HSV-1 human sera were analyzed. The average value of SPR response was 185 a.s. for negative sera and from 312 to. 1264 a.s. for positive sera. It was shown that SPR data were agreed with ELISA results in 96% of samples proving the great potential of SPR in such researches. It was investigated the possibility of biochip regeneration and it was shown that application of 10 mM NaOH solution leads to rupture of intermolecular bonds. This allows reuse the chip several times. Thus, in this study biosensor chip for detection of specific antibodies to HSV1 was successfully developed expanding a range of diagnostic methods for this pathogen.

Keywords: biochip, herpes virus, SPR

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Authors: Okolie Pius Ifeanyi, Emerenini Emilymary Chima

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Aims: The study investigated the diversity and identities of Lactic Acid Bacteria (LAB) isolated from different fresh fruits using Molecular Nested PCR analysis and the efficacy of cell free supernatants from Lactic Acid Bacteria (LAB) isolated from fresh fruits for in vitro control of some tomato pathogens. Study Design: Nested PCR approach was used in this study employing universal 16S rRNA gene primers in the first round PCR and LAB specific Primers in the second round PCR with the view of generating specific Nested PCR products for the LAB diversity present in the samples. The inhibitory potentials of supernatant obtained from LAB isolates of fruits origin that were molecularly characterized were investigated against some tomato phytopathogens using agar-well method with the view to develop biological agents for some tomato disease causing organisms. Methodology: Gram positive, catalase negative strains of LAB were isolated from fresh fruits on Man Rogosa and Sharpe agar (Lab M) using streaking method. Isolates obtained were molecularly characterized by means of genomic DNA extraction kit (Norgen Biotek, Canada) method. Standard methods were used for Nested Polymerase Chain Reaction (PCR) amplification targeting the 16S rRNA gene using universal 16S rRNA gene and LAB specific primers, agarose gel electrophoresis, purification and sequencing of generated Nested PCR products (Macrogen Inc., USA). The partial sequences obtained were identified by blasting in the non-redundant nucleotide database of National Center for Biotechnology Information (NCBI). The antimicrobial activities of characterized LAB against some tomato phytopathogenic bacteria which include (Xanthomonas campestries, Erwinia caratovora, and Pseudomonas syringae) were obtained by using the agar well diffusion method. Results: The partial sequences obtained were deposited in the database of National Centre for Biotechnology Information (NCBI). Isolates were identified based upon the sequences as Weissella cibaria (4, 18.18%), Weissella confusa (3, 13.64%), Leuconostoc paramensenteroides (1, 4.55%), Lactobacillus plantarum (8, 36.36%), Lactobacillus paraplantarum (1, 4.55%) and Lactobacillus pentosus (1, 4.55%). The cell free supernatants of LAB from fresh fruits origin (Weissella cibaria, Weissella confusa, Leuconostoc paramensenteroides, Lactobacillus plantarum, Lactobacillus paraplantarum and Lactobacillus pentosus) can inhibits these bacteria by creating clear zones of inhibition around the wells containing cell free supernatants of the above mentioned strains of lactic acid bacteria. Conclusion: This study shows that potentially LAB can be quickly characterized by molecular methods to specie level by nested PCR analysis of the bacteria isolate genomic DNA using universal 16S rRNA primers and LAB specific primer. Tomato disease causing organisms can be most likely biologically controlled by using extracts from LAB. This finding will reduce the potential hazard from the use of chemical herbicides on plant.

Keywords: nested pcr, molecular characterization, 16s rRNA gene, lactic acid bacteria

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Authors: Istvan Szilagyi, Marko Pavlovic, Paul Rouster

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Antioxidant enzymes are the most efficient defense systems against reactive oxygen species, which cause severe damage in living organisms and industrial products. However, their supplementation is problematic due to their high sensitivity to the environmental conditions. Immobilization on carrier nanoparticles is a promising research direction towards the improvement of their functional and colloidal stability. In that way, their applications in biomedical treatments and manufacturing processes in the food, textile and cosmetic industry can be extended. The main goal of the present research was to prepare and formulate antioxidant bionanocomposites composed of superoxide dismutase (SOD) enzyme, anionic clay (layered double hydroxide, LDH) nanoparticle and heparin (HEP) polyelectrolyte. To characterize the structure and the colloidal stability of the obtained compounds in suspension and solid state, electrophoresis, dynamic light scattering, transmission electron microscopy, spectrophotometry, thermogravimetry, X-ray diffraction, infrared and fluorescence spectroscopy were used as experimental techniques. LDH-SOD composite was synthesized by enzyme immobilization on the clay particles via electrostatic and hydrophobic interactions, which resulted in a strong adsorption of the SOD on the LDH surface, i.e., no enzyme leakage was observed once the material was suspended in aqueous solutions. However, the LDH-SOD showed only limited resistance against salt-induced aggregation and large irregularly shaped clusters formed during short term interval even at lower ionic strengths. Since sufficiently high colloidal stability is a key requirement in most of the applications mentioned above, the nanocomposite was coated with HEP polyelectrolyte to develop highly stable suspensions of primary LDH-SOD-HEP particles. HEP is a natural anticoagulant with one of the highest negative line charge density among the known macromolecules. The experimental results indicated that it strongly adsorbed on the oppositely charged LDH-SOD surface leading to charge inversion and to the formation of negatively charged LDH-SOD-HEP. The obtained hybrid materials formed stable suspension even under extreme conditions, where classical colloid chemistry theories predict rapid aggregation of the particles and unstable suspensions. Such a stabilization effect originated from electrostatic repulsion between the particles of the same sign of charge as well as from steric repulsion due to the osmotic pressure raised during the overlap of the polyelectrolyte chains adsorbed on the surface. In addition, the SOD enzyme kept its structural and functional integrity during the immobilization and coating processes and hence, the LDH-SOD-HEP bionanocomposite possessed excellent activity in decomposition of superoxide radical anions, as revealed in biochemical test reactions. In conclusion, due to the improved colloidal stability and the good efficiency in scavenging superoxide radical ions, the developed enzymatic system is a promising antioxidant candidate for biomedical or other manufacturing processes, wherever the aim is to decompose reactive oxygen species in suspensions.

Keywords: clay, enzyme, polyelectrolyte, formulation

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Authors: Olufunmilola A. Abiodun, Adegbola O. Dauda, Ayobami Ojo, Samson A. Oyeyinka

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Blighia sapida (Ackee) seed is a neglected and under-utilised crop. The fruit is cultivated for the aril which is used as meat substitute in soup while the seed is discarded. The seed is toxic due to the presence of hypoglycin which causes vomiting and death. The seed is shining black and bigger than the legume seeds. The seed contains high starch content which could serve as a cheap source of starch hereby reducing wastage of the crop during its season. Native starch had limitation in their use; therefore, modification of starch had been reported to improve the functional properties of starches. Therefore, this work determined the effect of modification on the properties of Blighia sapida seed starch. Blighia sapida seed was dehulled manually, milled and the starch extracted using standard method. The starch was subjected to modification using four methods (acid, alkaline, oxidized and acetylated methods). The morphological structure, form factor, granule size, amylose, swelling power, hypoglycin and pasting properties of the starches were determined. The structure of Blighia sapida using light microscope showed that the seed starch demonstrated an oval, round, elliptical, dome-shaped and also irregular shape. The form factors of the starch ranged from 0.32-0.64. Blighia sapida seed starches were smaller in granule sizes ranging from 2-6 µm. Acid modified starch had the highest amylose content (24.83%) and was significantly different ( < 0.05) from other starches. Blighia sapida seed starches showed a progressive increase in swelling power as temperature increased in native, acidified, alkalized, oxidized and acetylated starches but reduced with increasing temperature in pregelatinized starch. Hypoglycin A ranged from 3.89 to 5.74 mg/100 g with pregelatinized starch having the lowest value and alkalized starch having the highest value. Hypoglycin B ranged from 7.17 to 8.47 mg/100 g. Alkali-treated starch had higher peak viscosity (3973 cP) which was not significantly different (p > 0.05) from the native starch. Alkali-treated starch also was significantly different (p > 0.05) from other starches in holding strength value while acetylated starch had higher breakdown viscosity (1161.50 cP). Native starch was significantly different (p > 0.05) from other starches in final and setback viscosities. Properties of Blighia sapida modified starches showed that it could be used as a source of starch in food and other non-food industries and the toxic compound found in the starch was very low when compared to lethal dosage.

Keywords: Blighia sapida seed, modification, starch, hypoglycin

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Authors: Su Mon Saw, Joe Rothnagel

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Short open reading frames (sORFs) located in 5’UTR of mRNAs are known as uORFs. Characterization of uORF-encoded peptides (uPEPs) i.e., a subset of short open reading frame encoded peptides (sPEPs) and their translation regulation lead to understanding of causes of genetic disease, proteome complexity and development of treatments. Existence of uORFs within cellular proteome could be detected by LC-MS/MS. The ability of uORF to be translated into uPEP and achievement of uPEP identification will allow uPEP’s characterization, structures, functions, subcellular localization, evolutionary maintenance (conservation in human and other species) and abundance in cells. It is hypothesized that a subset of sORFs are translatable and that their encoded sPEPs are functional and are endogenously expressed contributing to the eukaryotic cellular proteome complexity. This project aimed to investigate whether sORFs encode functional peptides. Liquid chromatography-mass spectrometry (LC-MS) and bioinformatics were thus employed. Due to probable low abundance of sPEPs and small in sizes, the need for efficient peptide enrichment strategies for enriching small proteins and depleting the sub-proteome of large and abundant proteins is crucial for identifying sPEPs. Low molecular weight proteins were extracted using SDS-PAGE from Human Embryonic Kidney (HEK293) cells and Strong Cation Exchange Chromatography (SCX) from secreted HEK293 cells. Extracted proteins were digested by trypsin to peptides, which were detected by LC-MS/MS. The MS/MS data obtained was searched against Swiss-Prot using MASCOT version 2.4 to filter out known proteins, and all unmatched spectra were re-searched against human RefSeq database. ProteinPilot v5.0.1 was used to identify sPEPs by searching against human RefSeq, Vanderperre and Human Alternative Open Reading Frame (HaltORF) databases. Potential sPEPs were analyzed by bioinformatics. Since SDS PAGE electrophoresis could not separate proteins <20kDa, this could not identify sPEPs. All MASCOT-identified peptide fragments were parts of main open reading frame (mORF) by ORF Finder search and blastp search. No sPEP was detected and existence of sPEPs could not be identified in this study. 13 translated sORFs in HEK293 cells by mass spectrometry in previous studies were characterized by bioinformatics. Identified sPEPs from previous studies were <100 amino acids and <15 kDa. Bioinformatics results showed that sORFs are translated to sPEPs and contribute to proteome complexity. uPEP translated from uORF of SLC35A4 was strongly conserved in human and mouse while uPEP translated from uORF of MKKS was strongly conserved in human and Rhesus monkey. Cross-species conserved uORFs in association with protein translation strongly suggest evolutionary maintenance of coding sequence and indicate probable functional expression of peptides encoded within these uORFs. Translation of sORFs was confirmed by mass spectrometry and sPEPs were characterized with bioinformatics.

Keywords: bioinformatics, HEK293 cells, liquid chromatography-mass spectrometry, ProteinPilot, Strong Cation Exchange Chromatography, SDS-PAGE, sPEPs

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Authors: Murad Ali, Shoji Arai, Mohamed Khedr, Mukhtar Nasher, Shawki Nasr

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The Wadi Bayhan area of southeastern Yemen is about 60 km NW of Al-Bayda city in the Al-Bayda uplift terrane at the southeast margin of the Arabian-Nubian Shield. Intrusion of alkali gabbro into carbonate rocks apparently produced an 8m to 10 m thick reaction zone at the contact. This had been identified as nepheline pyroxenite. We have observed this to be mineralogically zoned with calc-silicate assemblages (e.g. pyroxene, calcite, spinel, garnet and melilite). The presence of melilite implies a skarn. The sinuous embayed pyroxenite-skarn contact, the presence of skarn minerals in pyroxenite, and textural evidence for growth of calc-silicate skarn by replacement of both carbonate rocks and solid pyroxenite indicate that reaction involved assimilation of carbonate wall rock by magma and loss of Al and Si to the skarn. Textural relationships between minerals provide evidence for a metasomatic development of the skarn at the expense of the pyroxenite. This process, related to the circulation of fluids equilibrated with carbonates, is responsible for those pyroxenite-spinel (± calcite) skarns. The uneven modal distribution of euhedral pyroxenite and enveloping nepheline in pyroxenite, the restricted occurrence of alkali gabbro as dikes in pyroxenite and skarn and the leucocratic matrix of pyroxenite suggest that pyroxenite represents an accumulation of titanaugite cemented by an alkali-rich residual magma and that alkali gabbro represents a part of the residual contaminated magma that was squeezed out of the pyroxene crystal mush. Carbonate assimilation is modeled by reaction of calcite and magmatic plagioclase, which results in resorption of plagioclase, growth of pyroxene enriched in Ca, Fe, Ti, and Al, and solution of nepheline in residual contaminated magma. The composition of nepheline pyroxenite evolved by addition of Ca from dissolved carbonate rocks, loss of Al and Si to skarn, and local segregation of solid pyroxene and alkali gabbro magma. The predominance of pyroxenite among contaminated rocks and their restriction to a large zone along the intrusive contact provide little evidence for the genesis of a significant volume of alkaline magmatic surroundings by carbonate assimilation.

Keywords: Yemen, Wadi Bayhan, skarn, pyroxenite, carbonatite, metasomatic

Procedia PDF Downloads 313

Authors: Pravina Gurjar, Bothiraja Pour, Vijay Kumbhar, Ganesh Dama

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Andrographolide (AG), a bioactive phytoconstituent, has a wider range of pharmacological action. However, due to the intestinal degradation, shows low oral bioavailability. The aim of the present work was to develop Floating In-situ gelling Gastro retentive System (FISGS) for AG in order to enhance its site specific absorption and minimize pH dependent hydrolysis in alkaline environment. Further to increase its therapeutic efficacy for peptic ulcer disease caused by H. pyroli. Gellan based floating in situ gelling system of AG were prepared by using sodium citrate and calcium carbonate. The 32 factorial designs was used to study the effect of gellan and calcium carbonate concentration (independent variables) on dependent variable such as viscosity, floating lag time and drug release. Developed system was evaluated for drug content, floating lag time, viscosity, and drug release studies. Drug content, viscosity, and floating lag time was found to be 81-99%, 67-117 Cps, and 3-5 sec, respectively. The obtained system showed good in vitro floating ability for more than 12 h using 0.1 N HCl as dissolution medium with initial burst release followed by the controlled zero order drug release up to 24 hrs. In vivo testing of FISGS of AG to rats demonstrated significant antiulcer activity that were evaluated by various parameters like pH, volume, total acidity, millimole equivalent of H+ ions/30 min, and protein content of gastric content. The densities of all the formulation batches were found to be near about 0.9 and floating duration above 12 hr. It was observed that with the increase in conc. of gellan there was increase in the viscosity of formulation but all formulations were in optimum range. The drug content of optimized batch was found to be 99.23. In histopathology study of stomach, the villi at the mucosal surface, the intercellular junction, the intestinal lumen were intact; no destruction of the epithelium, and submucosal gland in formulation treated and control group animals as compared to pure drug AG and standard ranitidine. Gellan-based in situ gastro retentive floating system could be advantageous in terms of increased bioavailability of AG to maintain an effective drug conc. in gastric fluid as well as in serum for longer period of time.

Keywords: andrographolide, floating drug delivery, in situ gelling system, gastroretentive system

Procedia PDF Downloads 346

Authors: Hamada Ahmed, Mervat A. Abdel-Latif, Alaa. A. Ghoraba, Samah A. Ganna

Abstract:

An experiment was conducted to determine the effect of microbial phytase (Quantum Blue®) supplementation on layer chickens fed diets with required or low phosphorous level in corn-soybean based diets. One hundred and sixteen 23-week-old Lohman brown laying hens were used in 8-week feeding trial. Hens were randomly allotted into four treatments where the group (1) (control group) was fed basal diet without phytase, group (2) fed basal diet supplemented with phytase, group (3) fed diet supplemented with phytase as a replacement of 25% of monocalcium phosphate and group (4) fed diet supplemented with phytase as a replacement of 50% of monocalcium phosphate. Records on daily egg production, egg mass, egg weight and body weight of hens at the end of experimental period were recorded. Results revealed no significant (p ≥ 0.05) differences were observed among the other dietary treatments in BW, egg production, egg mass, feed intake or feed conversion when these parameters were evaluated over the duration of the experiment while egg weight showed significant (p < 0.05) increase in all phytase supplemented groups. There was no significant (p ≥ 0.05) differences in egg quality including egg length, egg width, egg shape index, yolk height, yolk width, yolk index, yolk weight and yolk albumin ratio while egg albumin was significantly increased (p < 0.05) in group (2) and group (3). Egg shell weight increased significantly (p < 0.05) in all phytase supplemented groups when compared with the control group also shell thickness increased significantly (p < 0.05) in both group (2 &3). No significant (P ≥ 0.05) difference was observed in serum Ca, P level while alkaline phosphatase was significantly (P ˂ 0.05) increased in group (3). Egg shell analysis showed increase in egg shell ash% in all phytase supplemented groups when compared with the control group, egg shell calcium % was higher in group (3) and group (4) than the control group while group (2) showed lower egg shell calcium% than the other experimental groups, egg shell phosphorous% was higher in all phytase supplemented groups than the control group. Phosphorous digestability was significantly (P ˂ 0.05) increased in all phytase supplemented groups than the control group and the highest p digestability was in group (4). Calcium digestability showed significant (P ˂ 0.05) increase in all phytase supplemented groups when compared with the control group and the highest digetability was in group (4).

Keywords: layers, microbial phytase, Ca and P availability, egg production, egg characteristics

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