Search results for: Toxic protein
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 603

Search results for: Toxic protein

93 Nutritional Composition of Crackers Produced from Blend of Sprouted Pigeon Pea (Cajanus cajan), Unripe Plantain (Musa parasidiaca) and Brewers’ Spent Grain Flour and Blood Glucose Level of Diabetic Rats Fed the Biscuit

Authors: Nneka N. Uchegbu, Charles N. Ishiwu

Abstract:

The nutritional composition and hypoglycaemic effect of crackers produced from blend of sprouted pigeon pea, unripe plantain and brewers’ spent grain and fed to Alloxan induced diabetic rat was investigated. Crackers were produced from different blends of sprouted pigeon pea, unripe plantain and brewers’ spent grain. The crackers were evaluated for proximate composition, amino acid profile and antinutritional factors. Blood glucose levels of normal and diabetic rats fed with the control sample and different formulations of cracker were measured. The protein content of the samples were significantly different (p<0.05) from each other with sample A having the lowest value and sample B with the highest value. The values obtained showed that the samples contained most of the amino acids that are found in plant proteins. The levels of antinutritional factor determined were generally low. Administration of the formulated cracker meals led to a significant reduction in the fasting blood glucose level in the diabetic rats. The present study concluded that consumption of crackers produced from this composite flour could be recommended for the diabetics and those who are sceptical about the disease.

Keywords: Crackers, diabetics rat, sprouted pigeon pea, unripe plantain and brewers’ spent grain.

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92 Characteristic of Gluten-Free Products: Latvian Consumer Survey

Authors: Laila Ozola, Evita Straumite

Abstract:

Celiac disease is a permanent enteropathy caused by the ingestion of gluten, a protein occurring in wheat, rye and barley. The only way of the effective daily treatment is a strict gluten-free diet. From the investigation of products available in the local market, it was found that Latvian producers do not offer gluten-free products. The aim of this research was to study and analyze changes of celiac patient’s attitude to gluten-free product quality and availability in the Latvian market and purchasing habits. The survey was designed using website www.visidati.lv, and a questionnaire was sent to people suffering from celiac disease. The first time the respondents were asked to fill in the questionnaire in 2011, but now repeatedly from the beginning of September 2013 till the end of January 2014. The questionnaire was performed with 75 celiac patients, respondents were from all Latvian regions and they answered 16 questions. One of the most important questions was aimed to find out consumers’ opinion about quality of gluten-free products, consumption patterns of gluten-free products, and, moreover, their interest in products made in Latvia. Respondents were asked to name gluten-free products they mainly buy and give specific purchase locations, evaluate the quality of products and necessity for products produced in Latvia. The results of questionnaire show that the consumers are satisfied with the quality of gluten-free flour, flour blends, sweets and pasta, but are not satisfied with the quality of bread and confectionery available in the Latvian markets.

Keywords: Consumers, gluten-free products, quality, survey.

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91 Biological Effects of a Carbohydrate-Binding Protein from an Annelid, Perinereis nuntia Against Human and Phytopathogenic Microorganisms

Authors: Sarkar M. A. Kawsar, Sarkar M. A. Mamun, Md S. Rahman, Hidetaro Yasumitsu, Yasuhiro Ozeki

Abstract:

Lectins have a good scope in current clinical microbiology research. In the present study evaluated the antimicrobial activities of a D-galactose binding lectin (PnL) was purified from the annelid, Perinereis nuntia (polychaeta) by affinity chromatography. The molecular mass of the lectin was determined to be 32 kDa as a single polypeptide by SDS-PAGE under both reducing and non-reducing conditions. The hemagglutinating activity of the PnL showed against trypsinized and glutaraldehyde-fixed human erythrocytes was specifically inhibited by D-Gal, GalNAc, Galβ1-4Glc and Galα1-6Glc. PnL was evaluated for in vitro antibacterial screening studies against 11 gram-positive and gram-negative microorganisms. From the screening results, it was revealed that PnL exhibited significant antibacterial activity against gram-positive bacteria. Bacillus megaterium showed the highest growth inhibition by the lectin (250 μg/disc). However, PnL did not inhibit the growth of gram-negative bacteria such as Vibrio cholerae and Pseudomonas sp. PnL was also examined for in vitro antifungal activity against six fungal phytopathogens. PnL (100 μg/mL) inhibited the mycelial growth of Alternaria alternata (24.4%). These results indicate that future findings of lectin applications obtained from annelids may be of importance to life sciences.

Keywords: Perinereis nuntia, Lectin, Inhibition zone, Mycelial growth.

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90 Hypoglycemic Activity of Water Soluble Polysaccharides of Yam (Dioscorea hispida Dents) Prepared by Aqueous, Papain, and Tempeh Inoculum Assisted Extractions

Authors: Teti Estiasih, Harijono, Weny Bekti Sunarharum, Atina Rahmawati

Abstract:

This research studied the hypoglycemic effect of water soluble polysaccharide (WSP) extracted from yam (Dioscorea hispida) tuber by three different methods: aqueous extraction, papain assisted extraction, and tempeh inoculums assisted extraction. The two later extraction methods were aimed to remove WSP binding protein to have more pure WSP. The hypoglycemic activities were evaluated by means in vivo test on alloxan induced hyperglycemic rats, glucose response test (GRT), in situ glucose absorption test using everted sac, and short chain fatty acids (SCFAs) analysis. All yam WSP extracts exhibited ability to decrease blood glucose level in hyperglycemia condition as well as inhibited glucose absorption and SCFA formation. The order of hypoglycemic activity was tempeh inoculums assisted- >papain assisted- >aqueous WSP extracts. GRT and in situ glucose absorption test showed that order of inhibition was papain assisted- >tempeh inoculums assisted- >aqueous WSP extracts. Digesta of caecum of yam WSP extracts oral fed rats had more SCFA than control. Tempeh inoculums assisted WSP extract exhibited the most significant hypoglycemic activity.

Keywords: hypoglycemic activity, papain, tempeh inoculums, water soluble polysaccharides, yam (Discorea hispida)

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89 Processing and Assessment of Quality Characteristics of Composite Baby Foods

Authors: Reihaneh Ahmadzadeh Ghavidel, Mehdi Ghiafeh Davoodi

Abstract:

The usefulness of weaning foods to meet the nutrient needs of children is well recognized, and most of them are precooked roller dried mixtures of cereal and/or legume flours which posses a high viscosity and bulk when reconstituted. The objective of this study was to formulate composite weaning foods using cereals, malted legumes and vegetable powders and analyze them for nutrients, functional properties and sensory attributes. Selected legumes (green gram and lentil) were germinated, dried and dehulled. Roasted wheat, rice, carrot powder and skim milk powder also were used. All the ingredients were mixed in different proportions to get four formulations, made into 30% slurry and dried in roller drier. The products were analyzed for proximate principles, mineral content, functional and sensory qualities. The results of analysis showed following range of constituents per 100g of formulations on dry weight basis, protein, 18.1-18.9 g ; fat, 0.78-1.36 g ; iron, 5.09-6.53 mg; calcium, 265-310 mg. The lowest water absorption capacity was in case of wheat green gram based and the highest was in rice lentil based sample. Overall sensory qualities of all foods were graded as “good" and “very good" with no significant differences. The results confirm that formulated weaning foods were nutritionally superior, functionally appropriate and organoleptically acceptable.

Keywords: malted legumes, weaning foods, nutrition, functional properties

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88 The Acaricidal and Repellent Effect of Cinnamon Essential Oil against House Dust Mite

Authors: Je-Hoon Michael Oh

Abstract:

The major source of allergy in home is the house dust mite (Dematophagoides farina, Dermatophagoides pteronyssinus) causing allergic symptom include atopic dermatitis, asthma, perennial rhinitis and even infant death syndrome. Control of this mite species is dependent on the use of chemical methods such as fumigation treatments with methylene bromide, spraying with organophosphates such as pirimiphos-methyl, or treatments with repellents such as DEET and benzyl benzoate. Although effective, their repeated use for decades has sometimes resulted in development of resistance and fostered environmental and human health concerns. Both decomposing animal parts and the protein that surrounds mite fecal pellets cause mite allergy. So it is more effective to repel than to kill them because allergen is not living house dust mite but dead body or fecal particles of house dust mite. It is important to find out natural repellent material against house dust mite to control them and reduce the allergic reactions. Plants may be an alternative source for dust mite control because they contain a range of bioactive chemicals. The research objectives of this paper were to verify the acaricidal and repellent effects of cinnamon essential oil and to find out it-s most effective concentrations. We could find that cinnamon bark essential oil was very effective material to control the house dust mite. Furthermore, it could reduce chemical resistance and danger for human health.

Keywords: house dust mite, cinnamon, repellent effect

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87 The Influence of Organic Waste on Vegetable Nutritional Components and Healthy Livelihood, Minna, Niger State, Nigeria

Authors: A. Abdulkadir, A. A. Okhimamhe, Y. M. Bello, H. Ibrahim, D. H. Makun, M. T. Usman

Abstract:

Household waste form a larger proportion of waste generated across the state, accumulation of organic waste is an apparent problem and the existing dump sites could be overstress. Niger state has abundant arable land and water resources thus should be one of the highest producers of agricultural crops in the country. However, the major challenge to agricultural sector today is loss of soil nutrient coupled with high cost of fertilizer. These have continued to increase the use of fertilizer and decomposed solid waste for enhance agricultural yield, which have varying effects on the soil as well a threat to human livelihood. Consequently, vegetable yield samples from poultry droppings, decomposed household waste manure, NPK treatments and control from each replication were subjected to proximate analysis to determine the nutritional and antinutritional component as well as heavy metal concentration. Data collected was analyzed using SPSS software and Randomized complete Block Design means were compared. The result shows that the treatments do not devoid the concentrations of any nutritional components while the anti-nutritional analysis proved that NPK had higher oxalate content than control and organic treats. The concentration of lead and cadmium are within safe permissible level while the mercury level exceeded the FAO/WHO maximum permissible limit for the entire treatments depicts the need for urgent intervention to minimize mercury levels in soil and manure in order to mitigate its toxic effect. Thus, eco-agriculture should be widely accepted and promoted by the stakeholders for soil amendment, higher yield, strategies for sustainable environmental protection, food security, poverty eradication, attainment of sustainable development and healthy livelihood.

Keywords: Anti-nutritional, healthy livelihood, nutritional waste, organic waste.

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86 Evaluation of Storage Stability and Quality Parameters in Biscuit Made from Blends of Wheat, Cassava (Manihot esculenta) and Carrot (Daucus carota) Flour

Authors: Aminat. O Adelekan, Olawale T. Gbadebo

Abstract:

Biscuit is one of the most consumed cereal foods in Nigeria and research has shown that locally available tropical crops like cassava, sweet potato can be made into flour and used in the production of biscuits and other pastries. This study investigates some quality parameters in biscuits made from blends of wheat, cassava and carrot flour. The values of result of samples increased with increasing percentage substitution of cassava and carrot flour in some quality parameter like fiber, ash, gluten content, and carbohydrate. The protein content reduced significantly (P < 0.05) with increasing percentage substitution of cassava and carrot flour which ranged from 14.80% to 11.80% compared with the control sample which had 15.60%. There was a recorded significant increase (P < 0.05) in some mineral composition such as calcium, magnesium, sodium, iron, phosphorus, and vitamin A and C composition as the percentage substitution of cassava and carrot flour increased. During storage stability test, samples stored in the fridge and freezer were found to be the best storage location to preserve the sensory attributes and inhibit microbial growth when compared with storage under the sun and on the shelf. Biscuit made with blends of wheat, cassava and carrot flour can therefore serve as an alternative to biscuits made from 100% wheat flour, as they are richer in vitamin A, vitamin C, carbohydrate, dietary fiber and some essential minerals.

Keywords: Biscuit, carrot, flour blends, storage.

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85 Potential of Native Microorganisms in Tagus Estuary

Authors: Ana C. Sousa, Beatriz C. Santos, Fátima N. Serralha

Abstract:

The Tagus estuary is heavily affected by industrial and urban activities, making bioremediation studies crucial for environmental preservation. Fuel contamination in the area can arise from various anthropogenic sources, such as oil spills from shipping, fuel storage and transfer operations, and industrial discharges. These pollutants can cause severe harm to the ecosystem and the organisms, including humans, that inhabit it. Nonetheless, there are always natural organisms with the ability to resist these pollutants and transform them into non-toxic or harmless substances, which defines the process of bioremediation. Exploring the microbial communities existing in soil and their capacity to break down hydrocarbons has the potential to enhance the development of more efficient bioremediation approaches. The aim of this investigation was to explore the existence of hydrocarbonoclastic microorganisms in six locations within the Tagus estuary, three on the north bank: Trancão River, Praia Fluvial do Cais das Colinas and Praia de Algés, and three on the south bank: Praia Fluvial de Alcochete, Praia Fluvial de Alburrica, and Praia da Trafaria. In all studied locations, native microorganisms of the genus Pseudomonas were identified. The bioremediation rate of common hydrocarbons like gasoline, hexane, and toluene was assessed using the redox indicator 2,6-dichlorophenolindophenol (DCPIP). Effective hydrocarbon-degrading bacterial strains were identified in all analyzed areas, despite adverse environmental conditions. The highest bioremediation rates were achieved for gasoline (68%) in Alburrica, hexane (65%) in Algés, and toluene (79%) in Algés. Generally, the bacteria demonstrated efficient degradation of hydrocarbons added to the culture medium, with higher rates of aerobic biodegradation of hydrocarbons observed. These findings underscore the necessity for further in situ studies to better comprehend the relationship between native microbial communities and the potential for pollutant degradation in soil.

Keywords: Biodegradability rate, hydrocarbonoclastic microorganisms, soil bioremediation, Tagus estuary.

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84 VHL, PBRM1 and SETD2 Genes in Kidney Cancer: A Molecular Investigation

Authors: Rozhgar A. Khailany, Mehri Igci, Emine Bayraktar, Sakip Erturhan, Metin Karakok, Ahmet Arslan

Abstract:

Kidney cancer is the most lethal urological cancer accounting for 3% of adult malignancies. VHL, a tumor-suppressor gene, is best known to be associated with renal cell carcinoma (RCC). The VHL functions as negative regulator of hypoxia inducible factors. Recent sequencing efforts have identified several novel frequent mutations of histone modifying and chromatin remodeling genes in ccRCC (clear cell RCC) including PBRM1 and SETD2. The PBRM1 gene encodes the BAF180 protein, which involved in transcriptional activation and repression of selected genes. SETD2 encodes a histone methyltransferase, which may play a role in suppressing tumor development. In this study, RNAs of 30 paired tumor and normal samples that were grouped according to the types of kidney cancer and clinical characteristics of patients, including gender and average age were examined by RT-PCR, SSCP and sequencing techniques. VHL, PBRM1 and SETD2 expressions were relatively down-regulated. However, statistically no significance was found (Wilcoxon signed rank test, p>0.05). Interestingly, no mutation was observed on the contrary of previous studies. Understanding the molecular mechanisms involved in the pathogenesis of RCC has aided the development of molecular-targeted drugs for kidney cancer. Further analysis is required to identify the responsible genes rather than VHL, PBRM1 and SETD2 in kidney cancer.

Keywords: Kidney cancer, molecular biomarker, expression analysis, mutation screening.

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83 A Cross-Disciplinary Educational Model in Biomanufacturing to Sustain a Competitive Workforce Ecosystem

Authors: Rosa Buxeda, Lorenzo Saliceti-Piazza, Rodolfo J. Romañach, Luis Ríos, Sandra L. Maldonado-Ramírez

Abstract:

Biopharmaceuticals manufacturing is one of the major economic activities worldwide. Ninety-three percent of the workforce in a biomanufacturing environment concentrates in production-related areas. As a result, strategic collaborations between industry and academia are crucial to ensure the availability of knowledgeable workforce needed in an economic region to become competitive in biomanufacturing. In the past decade, our institution has been a key strategic partner with multinational biotechnology companies in supplying science and engineering graduates in the field of industrial biotechnology. Initiatives addressing all levels of the educational pipeline, from K-12 to college to continued education for company employees have been established along a ten-year span. The Amgen BioTalents Program was designed to provide undergraduate science and engineering students with training in biomanufacturing. The areas targeted by this educational program enhance their academic development, since these topics are not part of their traditional science and engineering curricula. The educational curriculum involved the process of producing a biomolecule from the genetic engineering of cells to the production of an especially targeted polypeptide, protein expression and purification, to quality control, and validation. This paper will report and describe the implementation details and outcomes of the first sessions of the program.

Keywords: Biomanufacturing curriculum, interdisciplinary learning, workforce development, industry-academia partnering.

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82 Simulation of Concrete Wall Subjected to Airblast by Developing an Elastoplastic Spring Model in Modelica Modelling Language

Authors: Leo Laine, Morgan Johansson

Abstract:

To meet the civilizations future needs for safe living and low environmental footprint, the engineers designing the complex systems of tomorrow will need efficient ways to model and optimize these systems for their intended purpose. For example, a civil defence shelter and its subsystem components needs to withstand, e.g. airblast and ground shock from decided design level explosion which detonates with a certain distance from the structure. In addition, the complex civil defence shelter needs to have functioning air filter systems to protect from toxic gases and provide clean air, clean water, heat, and electricity needs to also be available through shock and vibration safe fixtures and connections. Similar complex building systems can be found in any concentrated living or office area. In this paper, the authors use a multidomain modelling language called Modelica to model a concrete wall as a single degree of freedom (SDOF) system with elastoplastic properties with the implemented option of plastic hardening. The elastoplastic model was developed and implemented in the open source tool OpenModelica. The simulation model was tested on the case with a transient equivalent reflected pressure time history representing an airblast from 100 kg TNT detonating 15 meters from the wall. The concrete wall is approximately regarded as a concrete strip of 1.0 m width. This load represents a realistic threat on any building in a city like area. The OpenModelica model results were compared with an Excel implementation of a SDOF model with an elastic-plastic spring using simple fixed timestep central difference solver. The structural displacement results agreed very well with each other when it comes to plastic displacement magnitude, elastic oscillation displacement, and response times.

Keywords: Airblast from explosives, elastoplastic spring model, Modelica modelling language, SDOF, structural response of concrete structure.

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81 Investigating Prostaglandin E2 and Intracellular Oxidative Stress Levels in Lipopolysaccharide-Stimulated RAW 264.7 Macrophages upon Treatment with Strobilanthes crispus

Authors: Anna Pick Kiong Ling, Jia May Chin, Rhun Yian Koh, Ying Pei Wong

Abstract:

Background: Uncontrolled inflammation may cause serious inflammatory diseases if left untreated. Non-steroidal anti-inflammatory drug (NSAIDs) is commonly used to inhibit pro-inflammatory enzymes, thus, reduce inflammation. However, long term administration of NSAIDs leads to various complications. Medicinal plants are getting more attention as it is believed to be more compatible with human body. One of them is a flavonoid-containing medicinal plants, Strobilanthes crispus which has been traditionally claimed to possess anti-inflammatory and antioxidant activities. Nevertheless, its anti-inflammatory activities are yet to be scientifically documented. Objectives: This study aimed to examine the anti-inflammatory activity of S. crispus by investigating its effects on intracellular oxidative stress and prostaglandin E2 (PGE2) levels. Materials and Methods: In this study, the Maximum Non-toxic Dose (MNTD) of methanol extract of both leaves and stems of S. crispus was first determined using 3-(4,5-dimethylthiazolyl-2)-2,5-diphenytetrazolium Bromide (MTT) assay. The effects of S. crispus extracts at MNTD and half MNTD (½MNTD) on intracellular ROS as well as PGE2 levels in 1.0 µg/mL LPS-stimulated RAW 264.7 macrophages were then be measured using DCFH-DA and a competitive enzyme immunoassay kit, respectively. Results: The MNTD of leaf extract was determined as 700µg/mL while for stem was as low as 1.4µg/mL. When LPS-stimulated RAW 264.7 macrophages were subjected to the MNTD of S. crispus leaf extract, both intracellular ROS and PGE2 levels were significantly reduced. In contrast, stem extract at both MNTD and ½MNTD did not significantly reduce the PGE2 level, but significantly increased the intracellular ROS level. Conclusion: The methanol leaf extract of S. crispus may possess anti-inflammatory properties as it is able to significantly reduce the intracellular ROS and PGE2 levels of LPS-stimulated cells. Nevertheless, further studies such as investigating the interleukin, nitric oxide and cytokine tumor necrosis factor-α (TNFα) levels has to be conducted to further confirm the anti-inflammatory properties of S. crispus.

Keywords: Anti-inflammatory, natural products, prostaglandin E2, reactive oxygen species.

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80 Production and Purification of Monosaccharides by Hydrolysis of Sugar Cane Bagasse in an Ionic Liquid Medium

Authors: T. R. Bandara, H. Jaelani, G. J. Griffin

Abstract:

The conversion of lignocellulosic waste materials, such as sugar cane bagasse, to biofuels such as ethanol has attracted significant interest as a potential element for transforming transport fuel supplies to totally renewable sources. However, the refractory nature of the cellulosic structure of lignocellulosic materials has impeded progress on developing an economic process, whereby the cellulose component may be effectively broken down to glucose monosaccharides and then purified to allow downstream fermentation. Ionic liquid (IL) treatment of lignocellulosic biomass has been shown to disrupt the crystalline structure of cellulose thus potentially enabling the cellulose to be more readily hydrolysed to monosaccharides. Furthermore, conventional hydrolysis of lignocellulosic materials yields byproducts that are inhibitors for efficient fermentation of the monosaccharides. However, selective extraction of monosaccharides from an aqueous/IL phase into an organic phase utilizing a combination of boronic acids and quaternary amines has shown promise as a purification process. Hydrolysis of sugar cane bagasse immersed in an aqueous solution with IL (1-ethyl-3-methylimidazolium acetate) was conducted at different pH and temperature below 100 ºC. It was found that the use of a high concentration of hydrochloric acid to acidify the solution inhibited the hydrolysis of bagasse. At high pH (i.e. basic conditions), using sodium hydroxide, catalyst yields were reduced for total reducing sugars (TRS) due to the rapid degradation of the sugars formed. For purification trials, a supported liquid membrane (SLM) apparatus was constructed, whereby a synthetic solution containing xylose and glucose in an aqueous IL phase was transported across a membrane impregnated with phenyl boronic acid/Aliquat 336 to an aqueous phase. The transport rate of xylose was generally higher than that of glucose indicating that a SLM scheme may not only be useful for purifying sugars from undesirable toxic compounds, but also for fractionating sugars to improve fermentation efficiency.

Keywords: Biomass, bagasse, hydrolysis, monosaccharide, supported liquid membrane, purification.

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79 Evaluation of Hancornia speciosa Gomes Lyophilization at Different Stages of Maturation

Authors: D. C. Soares, J. T. S. Santos, D. G. Costa, A. K. S. Abud, T. P. Nunes, A. V. D. Figueiredo, A. M. de Oliveira Junior

Abstract:

Mangabeira (Hancornia speciosa Gomes), a native plant in Brazil, is found growing spontaneously in various regions of the country. The high perishability of tropical fruits such as mangaba, causes it to be necessary to use technologies that promote conservation, aiming to increase the shelf life of this fruit and add value. The objective of this study was to compare the mangabas lyophilization curves behaviors with different sizes and maturation stages. The fruits were freeze-dried for a period of approximately 45 hours at lyophilizer Liotop brand, model L -108. It has been considered large the fruits between 38 and 58 mm diameter and small, between 23 and 28 mm diameter and the two states of maturation, intermediate and mature. Large size mangabas drying curves in both states of maturation were linear behavior at all process, while the kinetic drying curves related to small fruits, independent of maturation state, had a typical behavior of drying, with all the well-defined steps. With these results it was noted that the time of lyophilization was suitable for small mangabas, a fact that did not happen with the larger one. This may indicate that the large mangabas require a longer time to freeze until reaches the equilibrium level, as it happens with the small fruits, going to have constant moisture at the end of the process. For both types of fruit were analyzed water activity, acidity, protein, lipid, and vitamin C before and after the process.

Keywords: Freeze dryer, mangaba, conservation, chemical characteristics.

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78 Biosensor Design through Molecular Dynamics Simulation

Authors: Wenjun Zhang, Yunqing Du, Steven W. Cranford, Ming L. Wang

Abstract:

The beginning of 21st century has witnessed new advancements in the design and use of new materials for biosensing applications, from nano to macro, protein to tissue. Traditional analytical methods lack a complete toolset to describe the complexities introduced by living systems, pathological relations, discrete hierarchical materials, cross-phase interactions, and structure-property dependencies. Materiomics – via systematic molecular dynamics (MD) simulation – can provide structureprocess- property relations by using a materials science approach linking mechanisms across scales and enables oriented biosensor design. With this approach, DNA biosensors can be utilized to detect disease biomarkers present in individuals’ breath such as acetone for diabetes. Our wireless sensor array based on single-stranded DNA (ssDNA)-decorated single-walled carbon nanotubes (SWNT) has successfully detected trace amount of various chemicals in vapor differentiated by pattern recognition. Here, we present how MD simulation can revolutionize the way of design and screening of DNA aptamers for targeting biomarkers related to oral diseases and oral health monitoring. It demonstrates great potential to be utilized to build a library of DNDA sequences for reliable detection of several biomarkers of one specific disease, and as well provides a new methodology of creating, designing, and applying of biosensors.

Keywords: Biosensor, design, DNA, molecular dynamics simulation.

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77 Digestibility in Yankasa Rams Fed Brachiaria ruziziensis – Centrosema pascuorum Hay Mixtures with Concentrate

Authors: Ibrahim Sani, J. T. Amodu, M. R. Hassan, R. J. Tanko, N. Adamu

Abstract:

This study investigated the digestibility of Brachiaria ruziziensis and Centrosema pascuorum hay mixtures at varying proportions in Yankasa rams. Twelve Yankasa rams with average initial weight 10.25 ± 0.1 kg were assigned to three dietary treatments of B. ruziziensis and C. pascuorum hay at different mixtures (75BR:25CP, 50BR:50CP and 25BR:75CP, respectively) in a Completely Randomized Design (CRD) for a period of 14 days. Concentrate diet was given to the experimental animals as supplement at fixed proportion, while the forage mixture (basal diet) was fed at 3% body weight. Animals on 50BR:50CP had better nutrient digestibility (crude protein, acid and neutral detergent fibre, ether extract and nitrogen free extract) than other treatment diets, except in dry matter digestibility (87.35%) which compared with 87.54% obtained in 25BR:75CP treatment diet and also organic matter digestibility. All parameters taken on nitrogen balance with the exception of nitrogen retained were significantly higher (P < 0.05) in animals fed 25BR:75CP diet, but were statistically similar with values obtained for animals on 50BR:50CP diet. From results obtained in this study, it is concluded that mixture of 25%BR75%CP gave the best nutrient digestibility and nitrogen balance in Yankasa rams. It is therefore recommended that B. ruziziensis and C. pascuorum should be fed at 50:50 mixture ratio for enhanced animal growth and performance in Nigeria.

Keywords: B. ruziziensis, C. pascuorum, digestibility, rams, Yankasa.

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76 New Coating Materials Based On Mixtures of Shellac and Pectin for Pharmaceutical Products

Authors: M. Kumpugdee-Vollrath, M. Tabatabaeifar, M. Helmis

Abstract:

Shellac is a natural polyester resin secreted by insects. Pectins are natural, non-toxic and water-soluble polysaccharides extracted from the peels of citrus fruits or the leftovers of apples. Both polymers are allowed for the use in the pharmaceutical industry and as a food additive. SSB Aquagold® is the aqueous solution of shellac and can be used for a coating process as an enteric or controlled drug release polymer. In this study, tablets containing 10 mg methylene blue as a model drug were prepared with a rotary press. Those tablets were coated with mixtures of shellac and one of the pectin different types (i.e. CU 201, CU 501, CU 701 and CU 020) mostly in a 2:1 ratio or with pure shellac in a small scale fluidized bed apparatus. A stable, simple and reproducible three-stage coating process was successfully developed. The drug contents of the coated tablets were determined using UV-VIS spectrophotometer. The characterization of the surface and the film thickness were performed with the scanning electron microscopy (SEM) and the light microscopy. Release studies were performed in a dissolution apparatus with a basket. Most of the formulations were enteric coated. The dissolution profiles showed a delayed or sustained release with a lagtime of at least 4 h. Dissolution profiles of coated tablets with pure shellac had a very long lagtime ranging from 13 to 17.9 h and the slopes were quite high. The duration of the lagtime and the slope of the dissolution profiles could be adjusted by adding the proper type of pectin to the shellac formulation and by variation of the coating amount. In order to apply a coating formulation as a colon delivery system, the prepared film should be resistant against gastric fluid for at least 2 h and against intestinal fluid for 4-6 h. The required delay time was gained with most of the shellac-pectin polymer mixtures. The release profiles were fitted with the modified model of the Korsmeyer-Peppas equation and the Hixson-Crowell model. A correlation coefficient (R²)> 0.99 was obtained by Korsmeyer-Peppas equation.

Keywords: Shellac, pectin, coating, fluidized bed, release, colon delivery system, kinetic, SEM, methylene blue.

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75 Lean Production to Increase Reproducibility and Work Safety in the Laser Beam Melting Process Chain

Authors: C. Bay, A. Mahr, H. Groneberg, F. Döpper

Abstract:

Additive Manufacturing processes are becoming increasingly established in the industry for the economic production of complex prototypes and functional components. Laser beam melting (LBM), the most frequently used Additive Manufacturing technology for metal parts, has been gaining in industrial importance for several years. The LBM process chain – from material storage to machine set-up and component post-processing – requires many manual operations. These steps often depend on the manufactured component and are therefore not standardized. These operations are often not performed in a standardized manner, but depend on the experience of the machine operator, e.g., levelling of the build plate and adjusting the first powder layer in the LBM machine. This lack of standardization limits the reproducibility of the component quality. When processing metal powders with inhalable and alveolar particle fractions, the machine operator is at high risk due to the high reactivity and the toxic (e.g., carcinogenic) effect of the various metal powders. Faulty execution of the operation or unintentional omission of safety-relevant steps can impair the health of the machine operator. In this paper, all the steps of the LBM process chain are first analysed in terms of their influence on the two aforementioned challenges: reproducibility and work safety. Standardization to avoid errors increases the reproducibility of component quality as well as the adherence to and correct execution of safety-relevant operations. The corresponding lean method 5S will therefore be applied, in order to develop approaches in the form of recommended actions that standardize the work processes. These approaches will then be evaluated in terms of ease of implementation and their potential for improving reproducibility and work safety. The analysis and evaluation showed that sorting tools and spare parts as well as standardizing the workflow are likely to increase reproducibility. Organizing the operational steps and production environment decreases the hazards of material handling and consequently improves work safety.

Keywords: Additive manufacturing, lean production, reproducibility, work safety.

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74 Adaptability of ‘Monti Dauni’ Bean Ecotypes in Plain Areas

Authors: Disciglio G., Nardella E., Gatta G., Giuliani M.M., Tarantino A.

Abstract:

The bean (Phaseolus vulgaris L.) is one of the best known of the legumes, and it has a long cultivation tradition in Italy. The territory of “Subappennino Dauno” (southern Italy) is at around 700 m a.s.l. and is predominantly grown with cereals, olive trees and grapevines. Ecotypes of white beans to eat dry (such as cannellini beans) are also grown, which are sought for their palatability, high digestibility, and ease of cooking. However, these are not easy to find on the market due to their low production in relatively small areas and on small family farms that use seeds handed down from generation to generation. The introduction of these ecotypes in plain areas of the Puglia region would provide an opportunity to promote the diffusion of this type of bean. To investigate the adaptability of these ecotypes in plain environments (Cerignola, in southern Italy) a comparative trial was carried out between three ‘Monti Dauni’ ecotypes (E1, E2, E3) that are native to mountain areas and the similar commercial variety, ‘Cannellini’. The data provide useful information about the quantitative and qualitative characteristics of these ecotypes when grown in lowland environments. Ecotype E3 provided the greatest bean production (2.34 t ha-1) compared to ‘Cannellini’ (1.28 t ha-1) and the other ecotypes (0.55 and 0.40 t ha-1, for E1 and E2, respectively), due to its greater plant growth and the larger size of the seed (and thickness, in particular). Finally, ecotype E2 provided the greatest protein content (31.2%), although not significantly different from the commercial cultivar ‘Cannellini’ (32.1%).

Keywords: 'Monti Dauni' bean, ecotypes, adaptability in plain areas, quali-quantitive.

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73 Effects of Process Parameters on the Yield of Oil from Coconut Fruit

Authors: Ndidi F. Amulu, Godian O. Mbah, Maxwel I. Onyiah, Callistus N. Ude

Abstract:

Analysis of the properties of coconut (Cocos nucifera) and its oil was evaluated in this work using standard analytical techniques. The analyses carried out include proximate composition of the fruit, extraction of oil from the fruit using different process parameters and physicochemical analysis of the extracted oil. The results showed the percentage (%) moisture, crude lipid, crude protein, ash and carbohydrate content of the coconut as 7.59, 55.15, 5.65, 7.35 and 19.51 respectively. The oil from the coconut fruit was odourless and yellowish liquid at room temperature (30oC). The treatment combinations used (leaching time, leaching temperature and solute: solvent ratio) showed significant differences (P<0.05) in the yield of oil from coconut flour. The oil yield ranged between 36.25%-49.83%. Lipid indices of the coconut oil indicated the acid value (AV) as 10.05Na0H/g of oil, free fatty acid (FFA) as 5.03%, saponification values (SV) as 183.26mgKOH-1g of oil, iodine value (IV) as 81.00 I2/g of oil, peroxide value (PV) as 5.00 ml/ g of oil and viscosity (V) as 0.002. A standard statistical package minitab version 16.0 program was used in the regression analysis and analysis of variance (ANOVA). The statistical software mentioned above was also used to generate various plots such as single effect plot, interactions effect plot and contour plot. The response or yield of oil from the coconut flour was used to develop a mathematical model that correlates the yield to the process variables studied. The maximum conditions obtained that gave the highest yield of coconut oil were leaching time of 2hrs, leaching temperature of 50oC and solute/solvent ratio of 0.05g/ml.

Keywords: Coconut, oil-extraction, optimization, physicochemical, proximate.

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72 Impact of Some Experimental Procedures on Behavioral Patterns and Physiological Traits of Rats

Authors: Amira, A. Goma, U. E. Mahrous

Abstract:

Welfare may be considered to be a subjective experience; it has a biological function that is related to the fitness and survival of the animal accordingly, researches have suggested that welfare is compromised when the animal's evolutionary fitness is reduced. This study was carried out to explain the effect of some managerial stressors as handling and restraint on behavioral patterns and biochemical parameters of rats. A total of 24 (12 males & 12 females) Sprague-Dawley rats (12 months & 150-180g) were allotted into 3 groups, handled group (4 male & 4 female), restrained group (4 male & 4 female) and control group (4 males & 4 females). The obtained results revealed that time spent feeding, drinking, movement and cage exploration frequencies increased significantly in handled rats than other groups, while lying time and licking increased significantly in restrained rats than handled and controls. Moreover, social behavior decreased in both stressed groups than control. Triglycerides were significantly increased in handled rats than other groups, while total lipid, total protein and globulin significantly increased in both treated groups than control. Corticosterone increased in restrained and handled rats than control ones. Moreover, there was an increment in packed cell volume significantly in restrained rats than others. These deducted that if we want to study the effect of stress on animal welfare it is necessary to study the effect of such stressors on animal’s behavior and physiological responses.

Keywords: Behavior, handling, restraint, rat, welfare.

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71 Experimental Investigation of the Impact of Biosurfactants on Residual-Oil Recovery

Authors: S. V. Ukwungwu, A. J. Abbas, G. G. Nasr

Abstract:

The increasing high price of natural gas and oil with attendant increase in energy demand on world markets in recent years has stimulated interest in recovering residual oil saturation across the globe. In order to meet the energy security, efforts have been made in developing new technologies of enhancing the recovery of oil and gas, utilizing techniques like CO2 flooding, water injection, hydraulic fracturing, surfactant flooding etc. Surfactant flooding however optimizes production but poses risk to the environment due to their toxic nature. Amongst proven records that have utilized other type of bacterial in producing biosurfactants for enhancing oil recovery, this research uses a technique to combine biosurfactants that will achieve a scale of EOR through lowering interfacial tension/contact angle. In this study, three biosurfactants were produced from three Bacillus species from freeze dried cultures using sucrose 3 % (w/v) as their carbon source. Two of these produced biosurfactants were screened with the TEMCO Pendant Drop Image Analysis for reduction in IFT and contact angle. Interfacial tension was greatly reduced from 56.95 mN.m-1 to 1.41 mN.m-1 when biosurfactants in cell-free culture (Bacillus licheniformis) were used compared to 4. 83mN.m-1 cell-free culture of Bacillus subtilis. As a result, cell-free culture of (Bacillus licheniformis) changes the wettability of the biosurfactant treatment for contact angle measurement to more water-wet as the angle decreased from 130.75o to 65.17o. The influence of microbial treatment on crushed rock samples was also observed by qualitative wettability experiments. Treated samples with biosurfactants remained in the aqueous phase, indicating a water-wet system. These results could prove that biosurfactants can effectively change the chemistry of the wetting conditions against diverse surfaces, providing a desirable condition for efficient oil transport in this way serving as a mechanism for EOR. The environmental friendly effect of biosurfactants applications for industrial purposes play important advantages over chemically synthesized surfactants, with various possible structures, low toxicity, eco-friendly and biodegradability.

Keywords: Bacillus, biosurfactant, enhanced oil recovery, residual oil, wettability.

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70 Implementation of a “DIVA“ Concept withspecific Elisa Kits; When Subunit H5 Avian Influenza Vaccine is used

Authors: Robles F, Uribe A, Guadarrama A , Castellanos L, González C.

Abstract:

The main objective of this study was to demonstrate that differentiation of infected and vaccinated animals (DIVA) strategy using different ELISA tests is possible when a subunit vaccine (Haemagglutinin protein) is used to prevent Avian influenza. Special emphasis was placed on the differentiation in the serological response to different components of the AIV (Nucleoprotein, Neuraminidase, Haemagglutinin, Nucleocapsid) between chickens that were vaccinated with a whole virus kill vaccine and recombinant vaccine. Furthermore, the potential use of this DIVA strategy using ELISA assays to detect Neuraminidase 1 (N1) was analyzed as strategy in countries where the field virus is H5N1 and the vaccine used is formulated with H5N2. Detection of AIV-s antibodies to any component in serum was negative for all animals on the study days 0-13. At study day 14 the titers of antibodies against Nucleoprotein (NP) and Nucleocapsid (NC) rose in the experimental groups vaccinated with Volvac® AI KV and were negatives during all the trial in the experimental groups vaccinated with a subunit H5; significant statistically differences were observed between these groups (p < 0.05). The seroconversion either Haemagglutinin or Neuraminidase was evident after 21 days post-vaccination in the experimental groups vaccinated with the respective viral fraction. Regarding the main aim of this study and according with the results that were obtained, use a combination of different ELISA test as a DIVA strategy is feasible when the vaccination is carry out with a subunit H5 vaccine. Also is possible to use the ELISA kit to detect Neuraminidase (either N1 or N2) as a DIVA concept in countries where H5N1 is present and the vaccination programs are done with H5N2 vaccine.

Keywords: Avian Influenza Virus, "DIVA concept", ELISAassay, subunit H5 vaccine.

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69 Differential Sensitivity of Nitrogen-Fixing, Filamentous Cyanobacterial Species to an Organochlorine Insecticide - 6, 7, 8, 9, 10, 10- Hexachloro-1, 5, 5a, 6, 9, 9a-Hexahydro-6, 9- Methano-2, 4, 3-Benzodioxathiepine-3-Oxide

Authors: Nirmal J.I. Kumar, Anubhuti A. Bora, Manmeet K. Amb

Abstract:

Application of pesticides in the paddy fields has deleterious effects on non-target organisms including cyanobacteria which are photosynthesizing and nitrogen fixing micro-organisms contributing significantly towards soil fertility and crop yield. Pesticide contamination in the paddy fields has manifested into a serious global environmental concern. To study the effect of one such pesticide, three cyanobacterial strains; Anabaena fertilissima, Aulosira fertilissima and Westiellopsis prolifica were selected for their stress responses to an Organochlorine insecticide - 6, 7, 8, 9, 10, 10-hexachloro-1, 5, 5a, 6, 9, 9a-hexahydro-6, 9-methano-2, 4, 3- benzodioxathiepine-3-oxide, with reference to their photosynthesic pigments-chlorophyll-a and carotenoids as well as accessory pigments-phycobiliproteins (phycocyanin, allophycocyanin and phycoerythrin), stress induced biochemical metabolites like carbohydrates, proteins, amino acids, phenols and enzymes-nitrate reductase, glutamine synthetase and succinate dehydrogenase. All the three cyanobacterial strains were adversely affected by the insecticide doses and inhibition was dose dependent. Reduction in photosynthetic and accessory pigments, metabolites, nitrogen fixing and respiratory enzymes of the test organisms were accompanied with an initial increase in their total protein at lower Organochlorine doses. On the other hand, increased amount of phenols in all the insecticide treated concentrations was indicative of stressed activities of the organisms.

Keywords: biochemical metabolites, endosulfan, enzymes, pigments

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68 Ellagic Acid Enhanced Apoptotic Radiosensitivity via G1 Cell Cycle Arrest and γ-H2AX Foci Formation in HeLa Cells in vitro

Authors: V. R. Ahire, A. Kumar, B. N. Pandey, K. P. Mishra, G. R. Kulkarni

Abstract:

Radiation therapy is an effective vital strategy used globally in the treatment of cervical cancer. However, radiation efficacy principally depends on the radiosensitivity of the tumor, and not all patient exhibit significant response to irradiation. A radiosensitive tumor is easier to cure than a radioresistant tumor which later advances to local recurrence and metastasis. Herbal polyphenols are gaining attention for exhibiting radiosensitization through various signaling. Current work focuses to study the radiosensitization effect of ellagic acid (EA), on HeLa cells. EA intermediated radiosensitization of HeLa cells was due to the induction γ-H2AX foci formation, G1 phase cell cycle arrest, and loss of reproductive potential, growth inhibition, drop in the mitochondrial membrane potential and protein expression studies that eventually induced apoptosis. Irradiation of HeLa in presence of EA (10 μM) to doses of 2 and 4 Gy γ-radiation produced marked tumor cytotoxicity. EA also demonstrated radio-protective effect on normal cell, NIH3T3 and aided recovery from the radiation damage. Our results advocate EA to be an effective adjuvant for improving cancer radiotherapy as it displays striking tumor cytotoxicity and reduced normal cell damage instigated by irradiation.

Keywords: Apoptotic radiosensitivity, ellagic acid, mitochondrial potential, cell-cycle arrest.

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67 Effect of Oral Administration of “Gadagi“ Tea on Liver Function in Rats

Authors: A. M. Gadanya, M. S. Sule, M. K. Atiku

Abstract:

Effect of oral administration of “Gadagi" tea on liver function was assessed on 50 healthy male albino rats which were grouped and administered with different doses(mg/kg) i.e low dose (380mg/kg, 415mg/kg, 365mg/kg, 315mg/kg for “sak", “sada" and “magani" respectively), standard dose ( 760mg/kg, 830mg/kg, 730mg/kg for “sak-, “sada" and “magani" respectively) and high dose (1500mg/kg, 1700mg/kg and 1460mg/kg for “sak--,"sada" and “magani" groups respectively) for a period of four weeks. Animals that were not administered with the tea constituted the control group. At the end of fourth week, the animals were sacrificed and their serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), total protein (TP), albumin (ALB), and globulins (GLO) were determined. Mean serum ALT and ALP activities were significantly higher (P<0.05) in rats orally administered with high dose of “sak" and those administered with standard dose of “sada" than those of the control group, suggesting a probable impairment of liver function due to liver cytolysis.Mean serum AST, ALT and ALP activities were significantly lower (P<0.05) in rats that were orally administered with high dose of “magani" than that of the control group, suggesting a probable improvement in liver function (due to decrease in liver cytolysis). Mean serum TP, ALB and GLO levels were significantly higher (P<0.05) in rats that were orally administered with the various doses of“sak", “sada" and “magani" than those of the control group. This also suggests a probable improvement in the synthetic function of the liver.Thus, some dosages of “sak" and “sada could be hepatotoxic, whereas “magani" especially at the high dose administered could have pharmacologically positive effect on the liver of the rats.

Keywords: Gadagi" tea, Liver function, Oral, Rats.

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66 Ligandless Extraction and Determination of Trace Amounts of Lead in Pomegranate, Zucchini and Lettuce Samples after Dispersive Liquid-Liquid Microextraction with Ultrasonic Bath and Optimization of Extraction Condition with RSM Design

Authors: Fariba Tadayon, Elmira Hassanlou, Hasan Bagheri, Mostafa Jafarian

Abstract:

Heavy metals are released into water, plants, soil, and food by natural and human activities. Lead has toxic roles in the human body and may cause serious problems even in low concentrations, since it may have several adverse effects on human. Therefore, determination of lead in different samples is an important procedure in the studies of environmental pollution. In this work, an ultrasonic assisted-ionic liquid based-liquid-liquid microextraction (UA-IL-DLLME) procedure for the determination of lead in zucchini, pomegranate, and lettuce has been established and developed by using flame atomic absorption spectrometer (FAAS). For UA-IL-DLLME procedure, 10 mL of the sample solution containing Pb2+ was adjusted to pH=5 in a glass test tube with a conical bottom; then, 120 μL of 1-Hexyl-3-methylimidazolium hexafluoro phosphate (CMIM)(PF6) was rapidly injected into the sample solution with a microsyringe. After that, the resulting cloudy mixture was treated by ultrasonic for 5 min, then the separation of two phases was obtained by centrifugation for 5 min at 3000 rpm and IL-phase diluted with 1 cc ethanol, and the analytes were determined by FAAS. The effect of different experimental parameters in the extraction step including: ionic liquid volume, sonication time and pH was studied and optimized simultaneously by using Response Surface Methodology (RSM) employing a central composite design (CCD). The optimal conditions were determined to be an ionic liquid volume of 120 μL, sonication time of 5 min, and pH=5. The linear ranges of the calibration curve for the determination by FAAS of lead were 0.1-4 ppm with R2=0.992. Under optimized conditions, the limit of detection (LOD) for lead was 0.062 μg.mL-1, the enrichment factor (EF) was 93, and the relative standard deviation (RSD) for lead was calculated as 2.29%. The levels of lead for pomegranate, zucchini, and lettuce were calculated as 2.88 μg.g-1, 1.54 μg.g-1, 2.18 μg.g-1, respectively. Therefore, this method has been successfully applied for the analysis of the content of lead in different food samples by FAAS.

Keywords: Dispersive liquid-liquid microextraction, Central composite design, Food samples, Flame atomic absorption spectrometry.

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65 The Role of Glutamine-Rich Region of Candida Albicans Tec1p in Mediating Morphological Transition and Invasive Growth

Authors: W. Abu Rayyan, A. Singh, A. M. Al-Jaafreh, W. Abu Dayyih, M. Bustami, S. Salem, N. Seder, K. Schröppel

Abstract:

Hyphal growth and the transcriptional regulation to the host environment are key issues during the pathogenesis of C. albicans. Tec1p is the C. albicans homolog of a TEA transcription factor family, which share a conserved DNA-binding TEA domain in their N-terminal. In order to define a structure-function relationship of the C. albicans Tec1p protein, we constructed several mutations on the N terminal, C terminal or in the TEA binding domain itself by homologous recombination technology. The modifications in the open reading frame of TEC1 were tested for reconstitution of the morphogenetic development of the tec1/tec1 mutant strain CaAS12. Mutation in the TEA consensus sequence did not confer transition to hyphae whereas the reconstitution of the full-length Tec1p has reconstituted hyphal development. A deletion in one of glutamine-rich regions either in the Tec1p N-terminal or the C-terminal in regions of 53-212 or 637–744 aa, respectively, did not restore morphological development in mutant CaAS12 strain. Whereas, the reconstitution with Tec1p mutants other than the glutamate-rich region has restored the morphogenetic switch. Additionally, the deletion of the glutamine-rich region has attenuated the invasive growth and the heat shock resistance of C. albicans. In conclusion, we show that a glutamine-rich region of Tec1p is essential for the hyphal development and mediating adaptation to the host environment of C. albicans.

Keywords: Candida albicans, transcription factor, TEA domain, hyphal formation, morphogenetic development, TEC1, Tet-induced.

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64 New Simultaneous High Performance Liquid Chromatographic Method for Determination of NSAIDs and Opioid Analgesics in Advanced Drug Delivery Systems and Human Plasma

Authors: Asad Ullah Madni, Mahmood Ahmad, Naveed Akhtar, Muhammad Usman

Abstract:

A new and cost effective RP-HPLC method was developed and validated for simultaneous analysis of non steroidal anti inflammatory dugs Diclofenac sodium (DFS), Flurbiprofen (FLP) and an opioid analgesic Tramadol (TMD) in advanced drug delivery systems (Liposome and Microcapsules), marketed brands and human plasma. Isocratic system was employed for the flow of mobile phase consisting of 10 mM sodium dihydrogen phosphate buffer and acetonitrile in molar ratio of 67: 33 with adjusted pH of 3.2. The stationary phase was hypersil ODS column (C18, 250×4.6 mm i.d., 5 μm) with controlled temperature of 30 C°. DFS in liposomes, microcapsules and marketed drug products was determined in range of 99.76-99.84%. FLP and TMD in microcapsules and brands formulation were 99.78 - 99.94 % and 99.80 - 99.82 %, respectively. Single step liquid-liquid extraction procedure using combination of acetonitrile and trichloroacetic acid (TCA) as protein precipitating agent was employed. The detection limits (at S/N ratio 3) of quality control solutions and plasma samples were 10, 20, and 20 ng/ml for DFS, FLP and TMD, respectively. The Assay was acceptable in linear dynamic range. All other validation parameters were found in limits of FDA and ICH method validation guidelines. The proposed method is sensitive, accurate and precise and could be applicable for routine analysis in pharmaceutical industry as well as in human plasma samples for bioequivalence and pharmacokinetics studies.

Keywords: Diclofenac Sodium, Flurbiprofen, Tramadol, HPLCUV detection, Validation.

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