Search results for: calcium fortification
Commenced in January 2007
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Edition: International
Paper Count: 762

Search results for: calcium fortification

42 Assessment of Groundwater Potential Sampled in Hand Dug Wells and Boreholes in Ado-Ekiti, Southwestern Nigeria

Authors: A. J. Olatunji, Adebolu Temitope Johnson

Abstract:

Groundwater samples were collected randomly from hand-dug wells and boreholes in parts of the Ado Ekiti metropolis and were subjected to quality assessment and characterization. Physicochemical analyses, which include the in-situ parameters (pH units, Turbidity, and Electrical Conductivity) and laboratory analysis of selected ionic concentrations, were carried out following standard methods. Hydrochemistry of the present study revealed relative mean concentrations of cations in the order Ca2+ > Na+ > Mg2+ > Cu2+> Fe > Mn2+ and that of anions: Cl- > NO3- > SO42- > F - respectively considering World Health Organisation Standard (WHO) range of values for potable water. The result shows that values of certain parameters (Total Dissolved Solid (TDS), Manganese, Calcium, Magnesium, Fluoride, and Sulphate) were below the Highest Desirable Level of the Standards, while values of some other parameters (pH Units, Electrical Conductivity, Turbidity, Alkalinity, Sodium, Copper, Chloride, and Total Hardness) were within the range of figures between Highest Desirable Level (HDL) and Maximum Permissible Level (MPL) of World Health Organization (WHO) drinking water Standards. The reduction in the mean concentration value of Total Dissolved Solids (TDS) of most borehole samples follows the fact that water had been allowed to settle in the overhead tanks before usage; we discussed and brainstormed in the course of sampling and agreed to take a sample that way because that represents what the people consume, it also shows an indication while there was slightly concentration increase of these soluble ions in hand-dug wells samples than borehole samples only with the exception of borehole sample seven BH7 because BH7 uses the mono-pumping system. These in-situ parameters and ionic concentrations were further displayed and or represented on bar charts along with the WHO standards for better pictorial clarifications. Deductions from field observation indices revealed the imprints of natural weathering, ion-exchange processes, and anthropogenic activities influencing groundwater quality. A strong degree of association was found to exist between sodium and chlorine ions in both hand-dug well and borehole groundwater samples through the use of Pearson’s correlation coefficient; this association can further be supported by the chemistry of the parent bedrock associated with the study area because the chemistry of groundwater is a replica of its host rock. The correlation of those two ions must have begun from the period of mountain building, indicating an identical source from which they were released to the groundwater. Moreover, considering the comparison of ionic species concentrations of all samples with the (WHO) standards, there were no anomalous increases or decreases in the laboratory analysis results; this simply reveals an insignificant state of pollution of the groundwater. The study and its sampling techniques were not set to target the likely area and extent of groundwater pollution but its portability. It could be said that the samples were safe for human consumption.

Keywords: groundwater, physicochemical, parameters ionic, concentrations, WHO standards

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41 Fodder Production and Livestock Rearing in Relation to Climate Change and Possible Adaptation Measures in Manaslu Conservation Area, Nepal

Authors: Bhojan Dhakal, Naba Raj Devkota, Chet Raj Upreti, Maheshwar Sapkota

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A study was conducted to find out the production potential, nutrient composition, and the variability of the most commonly available fodder trees along with the varying altitude to help optimize the dry matter requirement during winter lean period. The study was carried out from March to June, 2012 in Lho and Prok Village Development Committee of Manaslu Conservation Area (MCA), located in Gorkha district of Nepal. The other objective of the research was to learn the impact of climate change on livestock production linking it with feed availability. The study was conducted in two parts: social and biological. Accordingly, a households (HHs) survey was conducted to collect primary data from 70 HHs, focusing on the perception of respondents on impacts of climatic variability on the feeding management. The next part consisted of understanding yield potential and nutrient composition of the four most commonly available fodder trees (M. azedirach, M. alba, F. roxburghii, F. nemoralis), within two altitudes range: (1500-2000 masl and 2000-2500 masl) by using a RCB design in 2*4 factorial combination of treatments, each replicated four times. Results revealed that majority of the farmers perceived the change in climatic phenomenon more severely within the past five years. Farmers were using different adaptation technologies such as collection of forage from jungle, reducing unproductive animals, fodder trees utilization, and crop by product feeding at feed scarcity period. Ranking of the different fodder trees on the basis of indigenous knowledge and experiences revealed that F. roxburghii was the best-preferred fodder tree species (index value 0.72) in terms overall preferability whereas M. azedirach had highest growth and productivity (index value 0.77), F. roxburghii had highest adoptability (index value 0.69) and palatability (index value 0.69) as well. Similarly, fresh yield and dry matter yield of the each fodder trees was significant (P < 0.01) between the altitude and within species. Fodder trees yield analysis revealed that the highest dry matter (DM) yield (28 kg/tree) was obtained for F. roxburghii but that remained statistically similar (P > 0.05) to the other treatment. On the other hand, most of the parameters: ether extract (EE), acid detergent lignin (ADL), acid detergent fibre (ADF), cell wall digestibility (CWD), relative digestibility (RD), digestible nutrient (TDN), and Calcium (Ca) among the treatments were highly significant (P < 0.01). This indicates the scope of introducing productive and nutritive fodder trees species even at the high altitude to help reduce fodder scarcity problem during winter. The finding also revealed the scope of promoting all available local fodder trees species as crude protein content of these species were similar.

Keywords: fodder trees, yield potential, climate change, nutrient composition

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40 Impact of Material Chemistry and Morphology on Attrition Behavior of Excipients during Blending

Authors: Sri Sharath Kulkarni, Pauline Janssen, Alberto Berardi, Bastiaan Dickhoff, Sander van Gessel

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Blending is a common process in the production of pharmaceutical dosage forms where the high shear is used to obtain a homogenous dosage. The shear required can lead to uncontrolled attrition of excipients and affect API’s. This has an impact on the performance of the formulation as this can alter the structure of the mixture. Therefore, it is important to understand the driving mechanisms for attrition. The aim of this study was to increase the fundamental understanding of the attrition behavior of excipients. Attrition behavior of the excipients was evaluated using a high shear blender (Procept Form-8, Zele, Belgium). Twelve pure excipients are tested, with morphologies varying from crystalline (sieved), granulated to spray dried (round to fibrous). Furthermore, materials include lactose, microcrystalline cellulose (MCC), di-calcium phosphate (DCP), and mannitol. The rotational speed of the blender was set at 1370 rpm to have the highest shear with a Froude (Fr) number 9. Varying blending times of 2-10 min were used. Subsequently, after blending, the excipients were analyzed for changes in particle size distribution (PSD). This was determined (n = 3) by dry laser diffraction (Helos/KR, Sympatec, Germany). Attrition was found to be a surface phenomenon which occurs in the first minutes of the high shear blending process. An increase of blending time above 2 mins showed no change in particle size distribution. Material chemistry was identified as a key driver for differences in the attrition behavior between different excipients. This is mainly related to the proneness to fragmentation, which is known to be higher for materials such as DCP and mannitol compared to lactose and MCC. Secondly, morphology also was identified as a driver of the degree of attrition. Granular products consisting of irregular surfaces showed the highest reduction in particle size. This is due to the weak solid bonds created between the primary particles during the granulation process. Granular DCP and mannitol show a reduction of 80-90% in x10(µm) compared to a 20-30% drop for granular lactose (monohydrate and anhydrous). Apart from the granular lactose, all the remaining morphologies of lactose (spray dried-round, sieved-tomahawk, milled) show little change in particle size. Similar observations have been made for spray-dried fibrous MCC. All these morphologies have little irregular or sharp surfaces and thereby are less prone to fragmentation. Therefore, products containing brittle materials such as mannitol and DCP are more prone to fragmentation when exposed to shear. Granular products with irregular surfaces lead to an increase in attrition. While spherical, crystalline, or fibrous morphologies show reduced impact during high shear blending. These changes in size will affect the functionality attributes of the formulation, such as flow, API homogeneity, tableting, formation of dust, etc. Hence it is important for formulators to fully understand the excipients to make the right choices.

Keywords: attrition, blending, continuous manufacturing, excipients, lactose, microcrystalline cellulose, shear

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39 Biostratigraphic Significance of Shaanxilithes ningqiangensis from the Tal Group (Cambrian), Nigalidhar Syncline, Lesser Himalaya, India and Its GC-MS Analysis

Authors: C. A. Sharma, Birendra P. Singh

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We recovered 40 well preserved ribbon-shaped, meandering specimens of S. ningqiangensis from the Earthy Dolomite Member (Krol Group) and calcareous siltstone beds of the Earthy Siltstone Member (Tal Group) showing closely spaced annulations that lacked branching. The beginning and terminal points are indistinguishable. In certain cases, individual specimens are characterized by irregular, low-angle to high-angle sinuosity. It has been variously described as body fossil, ichnofossil and algae. Detailed study of this enigmatic fossil is needed to resolve the long standing controversy regarding its phylogenetic and stratigraphic placements, which will be an important contribution to the evolutionary history of metazoans. S. ningqiangensis has been known from the late Neoproterozoic (Ediacaran) of southern and central China (Sichuan, Shaanxi, Quinghai and Guizhou provinces and Ningxia Hui Autonomous region), Siberian platform and across Pc/C Boundary from latest Neoprterozoic to earliest Cambrian of northern India. Shaanxilithes is considered an Ediacaran organism that spans the Precambrian–Cambrian boundary, an interval marked by significant taphonomic and ecological transformations that include not only innovation but also probable extinction. All the past well constrained finds of S. ningqiangensis are restricted to Ediacaran age. However, due to the new recoveries of the fossil from Nigalidhar Syncline, the stratigraphic status of S. ningqiangensis-bearing Earthy Siltstone Member of the Shaliyan Formation of the Tal Group (Cambrian) is rendered uncertain, though the overlying Chert Member in the adjoining Korgai Syncline has yielded definite early Cambrian acritarchs. The moot question is whether the Earthy Siltstone Member represents an Ediacaran or an early Cambrian age?. It would be interesting to find if Shaanxilithes, so far known from Ediacaran sequences, could it transgress to the early Cambrian or in simple words could it withstand the Pc/C Boundary event? GC-MS data shows the S. ningqiangensis structure is formed by hydrocarbon organic compounds which are filled with inorganic elements filler like silica, Calcium, phosphorus etc. The S. ningqiangensis structure is a mixture of organic compounds of high molecular weight, containing several saturated rings with hydrocarbon chains having an occasional isolated carbon-carbon double bond and also containing, in addition, to small amounts of nitrogen, sulfur and oxygen. Data also revealed that the presence of nitrogen which would be either in the form of peptide chains means amide/amine or chemical form i.e. nitrates/nitrites etc. The formula weight and the weight ratio of C/H shows that it would be expected for algae derived organics, since algae produce fatty acids as well as other hydrocarbons such as cartenoids.

Keywords: GC-MS Analysis, lesser himalaya, Pc/C Boundary, shaanxilithes

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38 Treatment with Triton-X 100: An Enhancement Approach for Cardboard Bioprocessing

Authors: Ahlam Said Al Azkawi, Nallusamy Sivakumar, Saif Nasser Al Bahri

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Diverse approaches and pathways are under development with the determination to develop cellulosic biofuels and other bio-products eventually at commercial scale in “bio-refineries”; however, the key challenge is mainly the high level of complexity in processing the feedstock which is complicated and energy consuming. To overcome the complications in utilizing the naturally occurring lignocellulose biomass, using waste paper as a feedstock for bio-production may solve the problem. Besides being abundant and cheap, bioprocessing of waste paper has evolved in response to the public concern from rising landfill cost from shrinking landfill capacity. Cardboard (CB) is one of the major components of municipal solid waste and one of the most important items to recycle. Although 50-70% of cardboard constitute is known to be cellulose and hemicellulose, the presence of lignin around them cause hydrophobic cross-link which physically obstructs the hydrolysis by rendering it resistant to enzymatic cleavage. Therefore, pretreatment is required to disrupt this resistance and to enhance the exposure of the targeted carbohydrates to the hydrolytic enzymes. Several pretreatment approaches have been explored, and the best ones would be those can influence cellulose conversion rates and hydrolytic enzyme performance with minimal or less cost and downstream processes. One of the promising strategies in this field is the application of surfactants, especially non-ionic surfactants. In this study, triton-X 100 was used as surfactants to treat cardboard prior enzymatic hydrolysis and compare it with acid treatment using 0.1% H2SO4. The effect of the surfactant enhancement was evaluated through its effect on hydrolysis rate in respect to time in addition to evaluating the structural changes and modification by scanning electron microscope (SEM) and X-ray diffraction (XRD) and through compositional analysis. Further work was performed to produce ethanol from CB treated with triton-X 100 via separate hydrolysis and fermentation (SHF) and simultaneous saccharification and fermentation (SSF). The hydrolysis studies have demonstrated enhancement in saccharification by 35%. After 72 h of hydrolysis, a saccharification rate of 98% was achieved from CB enhanced with triton-X 100, while only 89 of saccharification achieved from acid pre-treated CB. At 120 h, the saccharification % exceeded 100 as reducing sugars continued to increase with time. This enhancement was not supported by any significant changes in the cardboard content as the cellulose, hemicellulose and lignin content remained same after treatment, but obvious structural changes were observed through SEM images. The cellulose fibers were clearly exposed with very less debris and deposits compared to cardboard without triton-X 100. The XRD pattern has also revealed the ability of the surfactant in removing calcium carbonate, a filler found in waste paper known to have negative effect on enzymatic hydrolysis. The cellulose crystallinity without surfactant was 73.18% and reduced to 66.68% rendering it more amorphous and susceptible to enzymatic attack. Triton-X 100 has proved to effectively enhance CB hydrolysis and eventually had positive effect on the ethanol yield via SSF. Treating cardboard with only triton-X 100 was a sufficient treatment to enhance the enzymatic hydrolysis and ethanol production.

Keywords: cardboard, enhancement, ethanol, hydrolysis, treatment, Triton-X 100

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37 Current Applications of Artificial Intelligence (AI) in Chest Radiology

Authors: Angelis P. Barlampas

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Learning Objectives: The purpose of this study is to inform briefly the reader about the applications of AI in chest radiology. Background: Currently, there are 190 FDA-approved radiology AI applications, with 42 (22%) pertaining specifically to thoracic radiology. Imaging findings OR Procedure details Aids of AI in chest radiology1: Detects and segments pulmonary nodules. Subtracts bone to provide an unobstructed view of the underlying lung parenchyma and provides further information on nodule characteristics, such as nodule location, nodule two-dimensional size or three dimensional (3D) volume, change in nodule size over time, attenuation data (i.e., mean, minimum, and/or maximum Hounsfield units [HU]), morphological assessments, or combinations of the above. Reclassifies indeterminate pulmonary nodules into low or high risk with higher accuracy than conventional risk models. Detects pleural effusion . Differentiates tension pneumothorax from nontension pneumothorax. Detects cardiomegaly, calcification, consolidation, mediastinal widening, atelectasis, fibrosis and pneumoperitoneum. Localises automatically vertebrae segments, labels ribs and detects rib fractures. Measures the distance from the tube tip to the carina and localizes both endotracheal tubes and central vascular lines. Detects consolidation and progression of parenchymal diseases such as pulmonary fibrosis or chronic obstructive pulmonary disease (COPD).Can evaluate lobar volumes. Identifies and labels pulmonary bronchi and vasculature and quantifies air-trapping. Offers emphysema evaluation. Provides functional respiratory imaging, whereby high-resolution CT images are post-processed to quantify airflow by lung region and may be used to quantify key biomarkers such as airway resistance, air-trapping, ventilation mapping, lung and lobar volume, and blood vessel and airway volume. Assesses the lung parenchyma by way of density evaluation. Provides percentages of tissues within defined attenuation (HU) ranges besides furnishing automated lung segmentation and lung volume information. Improves image quality for noisy images with built-in denoising function. Detects emphysema, a common condition seen in patients with history of smoking and hyperdense or opacified regions, thereby aiding in the diagnosis of certain pathologies, such as COVID-19 pneumonia. It aids in cardiac segmentation and calcium detection, aorta segmentation and diameter measurements, and vertebral body segmentation and density measurements. Conclusion: The future is yet to come, but AI already is a helpful tool for the daily practice in radiology. It is assumed, that the continuing progression of the computerized systems and the improvements in software algorithms , will redder AI into the second hand of the radiologist.

Keywords: artificial intelligence, chest imaging, nodule detection, automated diagnoses

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36 Cement Matrix Obtained with Recycled Aggregates and Micro/Nanosilica Admixtures

Authors: C. Mazilu, D. P. Georgescu, A. Apostu, R. Deju

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Cement mortars and concretes are some of the most used construction materials in the world, global cement production being expected to grow to approx. 5 billion tons, until 2030. But, cement is an energy intensive material, the cement industry being responsible for cca. 7% of the world's CO2 emissions. Also, natural aggregates represent non-renewable resources, exhaustible, which must be used efficiently. A way to reduce the negative impact on the environment is the use of additional hydraulically active materials, as a partial substitute for cement in mortars and concretes and/or the use of recycled concrete aggregates (RCA) for the recovery of construction waste, according to EU Directive 2018/851. One of the most effective active hydraulic admixtures is microsilica and more recently, with the technological development on a nanometric scale, nanosilica. Studies carried out in recent years have shown that the introduction of SiO2 nanoparticles into cement matrix improves the properties, even compared to microsilica. This is due to the very small size of the nanosilica particles (<100nm) and the very large specific surface, which helps to accelerate cement hydration and acts as a nucleating agent to generate even more calcium hydrosilicate which densifies and compacts the structure. The cementitious compositions containing recycled concrete aggregates (RCA) present, in generally, inferior properties compared to those obtained with natural aggregates. Depending on the degree of replacement of natural aggregate, decreases the workability of mortars and concretes with RAC, decrease mechanical resistances and increase drying shrinkage; all being determined, in particular, by the presence to the old mortar attached to the original aggregate from the RAC, which makes its porosity high and the mixture of components to require more water for preparation. The present study aims to use micro and nanosilica for increase the performance of some mortars and concretes obtained with RCA. The research focused on two types of cementitious systems: a special mortar composition used for encapsulating Low Level radioactive Waste (LLW); a composition of structural concrete, class C30/37, with the combination of exposure classes XC4+XF1 and settlement class S4. The mortar was made with 100% recycled aggregate, 0-5 mm sort and in the case of concrete, 30% recycled aggregate was used for 4-8 and 8-16 sorts, according to EN 206, Annex E. The recycled aggregate was obtained from a specially made concrete for this study, which after 28 days was crushed with the help of a Retsch jaw crusher and further separated by sieving on granulometric sorters. The partial replacement of cement was done progressively, in the case of the mortar composition, with microsilica (3, 6, 9, 12, 15% wt.), nanosilica (0.75, 1.5, 2.25% wt.), respectively mixtures of micro and nanosilica. The optimal combination of silica, from the point of view of mechanical resistance, was later used also in the case of the concrete composition. For the chosen cementitious compositions, the influence of micro and/or nanosilica on the properties in the fresh state (workability, rheological characteristics) and hardened state (mechanical resistance, water absorption, freeze-thaw resistance, etc.) is highlighted.

Keywords: cement, recycled concrete aggregates, micro/nanosilica, durability

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35 Storage of Organic Carbon in Chemical Fractions in Acid Soil as Influenced by Different Liming

Authors: Ieva Jokubauskaite, Alvyra Slepetiene, Danute Karcauskiene, Inga Liaudanskiene, Kristina Amaleviciute

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Soil organic carbon (SOC) is the key soil quality and ecological stability indicator, therefore, carbon accumulation in stable forms not only supports and increases the organic matter content in the soil, but also has a positive effect on the quality of soil and the whole ecosystem. Soil liming is one of the most common ways to improve the carbon sequestration in the soil. Determination of the optimum intensity and combinations of liming in order to ensure the optimal carbon quantitative and qualitative parameters is one of the most important tasks of this work. The field experiments were carried out at the Vezaiciai Branch of Lithuanian Research Centre for Agriculture and Forestry (LRCAF) during the 2011–2013 period. The effect of liming with different intensity (at a rate 0.5 every 7 years and 2.0 every 3-4 years) was investigated in the topsoil of acid moraine loam Bathygleyic Dystric Glossic Retisol. Chemical analyses were carried out at the Chemical Research Laboratory of Institute of Agriculture, LRCAF. Soil samples for chemical analyses were taken from the topsoil after harvesting. SOC was determined by the Tyurin method modified by Nikitin, measuring with spectrometer Cary 50 (VARIAN) at 590 nm wavelength using glucose standards. SOC fractional composition was determined by Ponomareva and Plotnikova version of classical Tyurin method. Dissolved organic carbon (DOC) was analyzed using an ion chromatograph SKALAR in water extract at soil-water ratio 1:5. Spectral properties (E4/E6 ratio) of humic acids were determined by measuring the absorbance of humic and fulvic acids solutions at 465 and 665 nm. Our study showed a negative statistically significant effect of periodical liming (at 0.5 and 2.0 liming rates) on SOC content in the soil. The content of SOC was 1.45% in the unlimed treatment, while in periodically limed at 2.0 liming rate every 3–4 years it was approximately by 0.18 percentage points lower. It was revealed that liming significantly decreased the DOC concentration in the soil. The lowest concentration of DOC (0.156 g kg-1) was established in the most intensively limed (2.0 liming rate every 3–4 years) treatment. Soil liming exerted an increase of all humic acids and fulvic acid bounded with calcium fractions content in the topsoil. Soil liming resulted in the accumulation of valuable humic acids. Due to the applied liming, the HR/FR ratio, indicating the quality of humus increased to 1.08 compared with that in unlimed soil (0.81). Intensive soil liming promoted the formation of humic acids in which groups of carboxylic and phenolic compounds predominated. These humic acids are characterized by a higher degree of condensation of aromatic compounds and in this way determine the intensive organic matter humification processes in the soil. The results of this research provide us with the clear information on the characteristics of SOC change, which could be very useful to guide the climate policy and sustainable soil management.

Keywords: acid soil, carbon sequestration, long–term liming, soil organic carbon

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34 Variability and Stability of Bread and Durum Wheat for Phytic Acid Content

Authors: Gordana Branković, Vesna Dragičević, Dejan Dodig, Desimir Knežević, Srbislav Denčić, Gordana Šurlan-Momirović

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Phytic acid is a major pool in the flux of phosphorus through agroecosystems and represents a sum equivalent to > 50% of all phosphorus fertilizer used annually. Nutrition rich in phytic acid can substantially decrease micronutrients apsorption as calcium, zink, iron, manganese, copper due to phytate salts excretion by human and non-ruminant animals as poultry, swine and fish, having in common very scarce phytase activity, and consequently the ability to digest and utilize phytic acid, thus phytic acid derived phosphorus in animal waste contributes to water pollution. The tested accessions consisted of 15 genotypes of bread wheat (Triticum aestivum L. ssp. vulgare) and of 15 genotypes of durum wheat (Triticum durum Desf.). The trials were sown at the three test sites in Serbia: Rimski Šančevi (RS) (45º19´51´´N; 19º50´59´´E), Zemun Polje (ZP) (44º52´N; 20º19´E) and Padinska Skela (PS) (44º57´N 20º26´E) during two vegetation seasons 2010-2011 and 2011-2012. The experimental design was randomized complete block design with four replications. The elementary plot consisted of 3 internal rows of 0.6 m2 area (3 × 0.2 m × 1 m). Grains were grinded with Laboratory Mill 120 Perten (“Perten”, Sweden) (particles size < 500 μm) and flour was used for the analysis. Phytic acid grain content was determined spectrophotometrically with the Shimadzu UV-1601 spectrophotometer (Shimadzu Corporation, Japan). Objectives of this study were to determine: i) variability and stability of the phytic acid content among selected genotypes of bread and durum wheat, ii) predominant source of variation regarding genotype (G), environment (E) and genotype × environment interaction (GEI) from the multi-environment trial, iii) influence of climatic variables on the GEI for the phytic acid content. Based on the analysis of variance it had been determined that the variation of phytic acid content was predominantly influenced by environment in durum wheat, while the GEI prevailed for the variation of the phytic acid content in bread wheat. Phytic acid content expressed on the dry mass basis was in the range 14.21-17.86 mg g-1 with the average of 16.05 mg g-1 for bread wheat and 14.63-16.78 mg g-1 with the average of 15.91 mg g-1 for durum wheat. Average-environment coordination view of the genotype by environment (GGE) biplot was used for the selection of the most desirable genotypes for breeding for low phytic acid content in the sense of good stability and lower level of phytic acid content. The most desirable genotypes of bread and durum wheat for breeding for phytic acid were Apache and 37EDUYT /07 No. 7849. Models of climatic factors in the highest percentage (> 91%) were useful in interpreting GEI for phytic acid content, and included relative humidity in June, sunshine hours in April, mean temperature in April and winter moisture reserves for genotypes of bread wheat, as well as precipitation in June and April, maximum temperature in April and mean temperature in June for genotypes of durum wheat.

Keywords: genotype × environment interaction, phytic acid, stability, variability

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33 Laboratory Assessment of Electrical Vertical Drains in Composite Soils Using Kaolin and Bentonite Clays

Authors: Maher Z. Mohammed, Barry G. Clarke

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As an alternative to stone column in fine grained soils, it is possible to create stiffened columns of soils using electroosmosis (electroosmotic piles). This program of this research is to establish the effectiveness and efficiency of the process in different soils. The aim of this study is to assess the capability of electroosmosis treatment in a range of composite soils. The combined electroosmotic and preloading equipment developed by Nizar and Clarke (2013) was used with an octagonal array of anodes surrounding a single cathode in a nominal 250mm diameter 300mm deep cylinder of soil and 80mm anode to cathode distance. Copper coiled springs were used as electrodes to allow the soil to consolidate either due to an external vertical applied load or electroosmosis. The equipment was modified to allow the temperature to be monitored during the test. Electroosmotic tests were performed on China Clay Grade E kaolin and calcium bentonite (Bentonex CB) mixed with sand fraction C (BS 1881 part 131) at different ratios by weight; (0, 23, 33, 50 and 67%) subjected to applied voltages (5, 10, 15 and 20). The soil slurry was prepared by mixing the dry soil with water to 1.5 times the liquid limit of the soil mixture. The mineralogical and geotechnical properties of the tested soils were measured before the electroosmosis treatment began. In the electroosmosis cell tests, the settlement, expelled water, variation of electrical current and applied voltage, and the generated heat was monitored during the test time for 24 osmotic tests. Water content was measured at the end of each test. The electroosmotic tests are divided into three phases. In Phase 1, 15 kPa was applied to simulate a working platform and produce a uniform soil which had been deposited as a slurry. 50 kPa was used in Phase 3 to simulate a surcharge load. The electroosmotic treatment was only performed during Phase 2 where a constant voltage was applied through the electrodes in addition to the 15 kPa pressure. This phase was stopped when no further water was expelled from the cell, indicating the electroosmotic process had stopped due to either the degradation of the anode or the flow due to the hydraulic gradient exactly balanced the electroosmotic flow resulting in no flow. Control tests for each soil mixture were carried out to assess the behaviour of the soil samples subjected to only an increase of vertical pressure, which is 15kPa in Phase 1 and 50kPa in Phase 3. Analysis of the experimental results from this study showed a significant dewatering effect on the soil slurries. The water discharged by the electroosmotic treatment process decreased as the sand content increased. Soil temperature increased significantly when electrical power was applied and drops when applied DC power turned off or when the electrode degraded. The highest increase in temperature was found in pure clays at higher applied voltage after about 8 hours of electroosmosis test.

Keywords: electrokinetic treatment, electrical conductivity, electroosmotic consolidation, electroosmosis permeability ratio

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32 Development of a Human Skin Explant Model for Drug Metabolism and Toxicity Studies

Authors: K. K. Balavenkatraman, B. Bertschi, K. Bigot, A. Grevot, A. Doelemeyer, S. D. Chibout, A. Wolf, F. Pognan, N. Manevski, O. Kretz, P. Swart, K. Litherland, J. Ashton-Chess, B. Ling, R. Wettstein, D. J. Schaefer

Abstract:

Skin toxicity is poorly detected during preclinical studies, and drug-induced side effects in humans such as rashes, hyperplasia or more serious events like bullous pemphigus or toxic epidermal necrolysis represent an important hurdle for clinical development. In vitro keratinocyte-based epidermal skin models are suitable for the detection of chemical-induced irritancy, but do not recapitulate the biological complexity of full skin and fail to detect potential serious side-effects. Normal healthy skin explants may represent a valuable complementary tool, having the advantage of retaining the full skin architecture and the resident immune cell diversity. This study investigated several conditions for the maintenance of good morphological structure after several days of culture and the retention of phase II metabolism for 24 hours in skin explants in vitro. Human skin samples were collected with informed consent from patients undergoing plastic surgery and immediately transferred and processed in our laboratory by removing the underlying dermal fat. Punch biopsies of 4 mm diameter were cultured in an air-liquid interface using transwell filters. Different cultural conditions such as the effect of calcium, temperature and cultivation media were tested for a period of 14 days and explants were histologically examined after Hematoxylin and Eosin staining. Our results demonstrated that the use of Williams E Medium at 32°C maintained the physiological integrity of the skin for approximately one week. Upon prolonged incubation, the upper layers of the epidermis become thickened and some dead cells are present. Interestingly, these effects were prevented by addition of EGFR inhibitors such as Afatinib or Erlotinib. Phase II metabolism of the skin such as glucuronidation (4-methyl umbeliferone), sulfation (minoxidil), N-acetyltransferase (p-toluidene), catechol methylation (2,3-dehydroxy naphthalene), and glutathione conjugation (chlorodinitro benzene) were analyzed by using LCMS. Our results demonstrated that the human skin explants possess metabolic activity for a period of at least 24 hours for all the substrates tested. A time course for glucuronidation with 4-methyl umbeliferone was performed and a linear correlation was obtained over a period of 24 hours. Longer-term culture studies will indicate the possible evolution of such metabolic activities. In summary, these results demonstrate that human skin explants maintain a normal structure for several days in vitro and are metabolically active for at least the first 24 hours. Hence, with further characterisation, this model may be suitable for the study of drug-induced toxicity.

Keywords: human skin explant, phase II metabolism, epidermal growth factor receptor, toxicity

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31 The Effects of Bisphosphonates on Osteonecrosis of Jaw Bone: A Stem Cell Perspective

Authors: Huseyin Apdik, Aysegul Dogan, Selami Demirci, Ezgi Avsar Apdik, Fikrettin Sahin

Abstract:

Mesenchymal stem cells (MSCs) are crucial cell types for bone maintenance and growth along with resident bone progenitor cells providing bone tissue integrity during osteogenesis and skeletal growth. Any deficiency in this regulation would result in vital bone diseases. Of those, osteoporosis, characterized by a reduction in bone mass and mineral density, is a critical skeletal disease for especially elderly people. The commonly used drugs for the osteoporosis treatment are bisphosphonates (BPs). The most prominent role of BPs is to prevent bone resorption arisen from high osteoclast activity. However, administrations of bisphosphonates may also cause bisphosphonate-induced osteonecrosis of the jaw (BIONJ). Up to the present, the researchers have proposed several circumstances for BIONJ. However, effects of long-term and/or high dose usage of BPs on stem cell’s proliferation, survival, differentiation or maintenance capacity have not been evaluated yet. The present study will be held to; figure out BPs’ effects on MSCs in vitro in the aspect of cell proliferation and toxicity, migration, angiogenic activity, lineage specific gene and protein expression levels, mesenchymal stem cell properties and potential signaling pathways affected by BP treatment. Firstly, mesenchymal stem cell characteristics of Dental Pulp Stem Cells (DPSCs) and Periodontal Ligament Stem Cells (PDLSCs) were proved using flow cytometry analysis. Cell viability analysis was completed to determine the cytotoxic effects of BPs (Zoledronate (Zol), Alendronate (Ale) and Risedronate (Ris)) on DPSCs and PDLSCs by the 3-(4,5-di-methyl-thiazol-2-yl)-5-(3-carboxymethoxy-phenyl)-2-(4-sulfo-phenyl)-2H-tetrazolium (MTS) assay. Non-toxic concentrations of BPs were determined at 24 h under growth condition, and at 21 days under osteogenic differentiation condition for both cells. The scratch assay was performed to evaluate their migration capacity under the usage of determined of BPs concentrations at 24 h. The results revealed that while the scratch closure is 70% in the control group for DPSCs, it was 57%, 66% and 66% in Zol, Ale and Ris groups, respectively. For PDLSs, while wound closure is 71% in control group, it was 65%, 66% and 66% in Zol, Ale and Ris groups, respectively. As future experiments, tube formation assay and aortic ring assay will be done to determinate angiogenesis abilities of DPSCs and PDLSCs treated with BPs. Expression levels of osteogenic differentiation marker genes involved in bone development will be determined using real time-polymerase change reaction (RT-PCR) assay and expression profiles of important proteins involved in osteogenesis will be evaluated using western blotting assay for osteogenically differentiated MSCs treated with or without BPs. In addition to these, von Kossa staining will be performed to measure calcium mineralization status of MSCs.

Keywords: bisphosphonates, bisphosphonate-induced osteonecrosis of the jaw, mesenchymal stem cells, osteogenesis

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30 Strength Properties of Ca-Based Alkali Activated Fly Ash System

Authors: Jung-Il Suh, Hong-Gun Park, Jae-Eun Oh

Abstract:

Recently, the use of long-span precast concrete (PC) construction has increased in modular construction such as storage buildings and parking facilities. When applying long span PC member, reducing weight of long span PC member should be conducted considering lifting capacity of crane and self-weight of PC member and use of structural lightweight concrete made by lightweight aggregate (LWA) can be considered. In the process of lightweight concrete production, segregation and bleeding could occur due to difference of specific gravity between cement (3.3) and lightweight aggregate (1.2~1.8) and reducing weight of binder is needed to prevent the segregation between binder and aggregate. Also, lightweight precast concrete made by cementitious materials such as fly ash and ground granulated blast furnace (GGBFS) which is lower than specific gravity of cement as a substitute for cement has been studied. When only using fly ash for cementless binder alkali-activation of fly ash is most important chemical process in which the original fly ash is dissolved by a strong alkaline medium in steam curing with high-temperature condition. Because curing condition is similar with environment of precast member production, additional process is not needed. Na-based chloride generally used as a strong alkali activator has a practical problem such as high pH toxicity and high manufacturing cost. Instead of Na-based alkali activator calcium hydroxide [Ca(OH)2] and sodium hydroxide [Na2CO3] might be used because it has a lower pH and less expensive than Na-based alkali activator. This study explored the influences on Ca(OH)2-Na2CO3-activated fly ash system in its microstructural aspects and strength and permeability using powder X-ray analysis (XRD), thermogravimetry (TGA), mercury intrusion porosimetry (MIP). On the basis of microstructural analysis, the conclusions are made as follows. Increase of Ca(OH)2/FA wt.% did not affect improvement of compressive strength. Also, Ca(OH)2/FA wt.% and Na2CO3/FA wt.% had little effect on specific gravity of saturated surface dry (SSD) and absolute dry (AD) condition to calculate water absorption. Especially, the binder is appropriate for structural lightweight concrete because specific gravity of the hardened paste has no difference with that of lightweight aggregate. The XRD and TGA/DTG results did not present considerable difference for the types and quantities of hydration products depending on w/b ratio, Ca(OH)2 wt.%, and Na2CO3 wt.%. In the case of higher molar quantity of Ca(OH)2 to Na2CO3, XRD peak indicated unreacted Ca(OH)2 while DTG peak was not presented because of small quantity. Thus, presence of unreacted Ca(OH)2 is too small quantity to effect on mechanical performance. As a result of MIP, the porosity volume related to capillary pore depends on the w/b ratio. In the same condition of w/b ratio, quantities of Ca(OH)2 and Na2CO3 have more influence on pore size distribution rather than total porosity. While average pore size decreased as Na2CO3/FA w.t% increased, the average pore size increased over 20 nm as Ca(OH)2/FA wt.% increased which has inverse proportional relationship between pore size and mechanical properties such as compressive strength and water permeability.

Keywords: Ca(OH)2, compressive strength, microstructure, fly ash, Na2CO3, water absorption

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29 Temporal Variation of Surface Runoff and Interrill Erosion in Different Soil Textures of a Semi-arid Region, Iran

Authors: Ali Reza Vaezi, Naser Fakori Ivand, Fereshteh Azarifam

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Interrill erosion is the detachment and transfer of soil particles between the rills due to the impact of raindrops and the shear stress of shallow surface runoff. This erosion can be affected by some soil properties such as texture, amount of organic matter and stability of soil aggregates. Information on the temporal variation of interrill erosion during a rainfall event and the effect soil properties have on it can help in understanding the process of runoff production and soil loss between the rills in hillslopes. The importance of this study is especially grate in semi-arid regions, where the soil is weakly aggregated and vegetation cover is mostly poor. Therefore, this research was conducted to investigate the temporal variation of surface flow and interrill erosion and the effect of soil properties on it in some semi-arid soils. A field experiment was done in eight different soil textures under simulated rainfalls with uniform intensity. A total of twenty four plots were installed for eight study soils with three replicates in the form of a random complete block design along the land. The plots were 1.2 m (length) × 1 m (width) in dimensions which designed with a distance of 3 m from each other across the slope. Then, soil samples were purred into the plots. The plots were surrounded by a galvanized sheet, and runoff and soil erosion equipment were placed at their outlets. Rainfall simulation experiments were done using a designed portable simulator with an intensity of 60 mm per hour for 60 minutes. A plastic cover was used around the rainfall simulator frame to prevent the impact of the wind on the free fall of water drops. Runoff production and soil loss were measured during 1 hour time with 5-min intervals. In order to study soil properties, such as particle size distribution, aggregate stability, bulk density, ESP and Ks were determined in the laboratory. Correlation and regression analysis was done to determine the effect of soil properties on runoff and interrill erosion. Results indicated that the study soils have lower booth organic matter content and aggregate stability. The soils, except for coarse textured textures, are calcareous and with relatively higher exchangeable sodium percentages (ESP). Runoff production and soil loss didn’t occur in sand, which was associated with higher infiltration and drainage rates. In other study soils, interrill erosion occurred simultaneously with the generation of runoff. A strong relationship was found between interrill erosion and surface runoff (R2 = 0.75, p< 0.01). The correlation analysis showed that surface runoff was significantly affected by some soil properties consisting of sand, silt, clay, bulk density, gravel, hydraulic conductivity (Ks), lime (calcium carbonate), and ESP. The soils with lower Ks such as fine-textured soils, produced higher surface runoff and more interrill erosion. In the soils, Surface runoff production temporally increased during rainfall and finally reached a peak after about 25-35 min. Time to peak was very short (30 min) in fine-textured soils, especially clay, which was related to their lower infiltration rate.

Keywords: erosion plot, rainfall simulator, soil properties, surface flow

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28 Exploring Bio-Inspired Catecholamine Chemistry to Design Durable Anti-Fungal Wound Dressings

Authors: Chetna Dhand, Venkatesh Mayandi, Silvia Marrero Diaz, Roger W. Beuerman, Seeram Ramakrishna, Rajamani Lakshminarayanan

Abstract:

Sturdy Insect Cuticle Sclerotization, Incredible Substrate independent Mussel’s bioadhesion, Tanning of Leather are some of catechol(amine)s mediated natural processes. Chemical contemplation spots toward a mechanism instigated with the formation of the quinone moieties from the respective catechol(amine)s, via oxidation, followed by the nucleophilic addition of the amino acids/proteins/peptides to this quinone leads to the development of highly strong, cross-linked and water-resistant proteinacious structures. Inspired with this remarkable catechol(amine)s chemistry towards amino acids/proteins/peptides, we attempted to design highly stable and water-resistant antifungal wound dressing mats with exceptional durability using collagen (protein), dopamine (catecholamine) and antifungal drugs (Amphotericin B and Caspofungin) as the key materials. Electrospinning technique has been used to fabricate desired nanofibrous mat including Collagen (COLL), COLL/Dopamine (COLL/DP) and calcium incorporated COLL/DP (COLL-DP-Ca2+). The prepared protein-based scaffolds have been studied for their microscopic investigations (SEM, TEM, and AFM), structural analysis (FT-IR), mechanical properties, water wettability characteristics and aqueous stability. Biocompatibility of these scaffolds has been analyzed for dermal fibroblast cells using MTS assay, Cell TrackerTM Green CMFDA and confocal imaging. Being the winner sample, COLL-DP-Ca2+ scaffold has been selected for incorporating two antifungal drugs namely Caspofungin (Peptide based) and Amphotericin B (Non-Peptide based). Antifungal efficiency of the designed mats has been evaluated for eight diverse fungal strains employing different microbial assays including disc diffusion, cell-viability assay, time kill kinetics etc. To confirm the durability of these mats, in term of their antifungal activity, drug leaching studies has been performed and monitored using disc diffusion assay each day. Ex-vivo fungal infection model has also been developed and utilized to validate the antifungal efficacy of the designed wound dressings. Results clearly reveal dopamine mediated crosslinking within COLL-antifungal scaffolds that leads to the generation of highly stable, mechanical tough, biocompatible wound dressings having the zone of inhabitation of ≥ 2 cm for almost all the investigated fungal strains. Leaching studies and Ex-vivo model has confirmed the durability of these wound dressing for more than 3 weeks and certified their suitability for commercialization. A model has also been proposed to enlighten the chemical mechanism involved for the development of these antifungal wound dressings with exceptional robustness.

Keywords: catecholamine chemistry, electrospinning technique, antifungals, wound dressings, collagen

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27 Biomaterials Solutions to Medical Problems: A Technical Review

Authors: Ashish Thakur

Abstract:

This technical paper was written in view of focusing the biomaterials and its various applications in modern industries. Author tires to elaborate not only the medical, infect plenty of application in other industries. The scope of the research area covers the wide range of physical, biological and chemical sciences that underpin the design of biomaterials and the clinical disciplines in which they are used. A biomaterial is now defined as a substance that has been engineered to take a form which, alone or as part of a complex system, is used to direct, by control of interactions with components of living systems, the course of any therapeutic or diagnostic procedure. Biomaterials are invariably in contact with living tissues. Thus, interactions between the surface of a synthetic material and biological environment must be well understood. This paper reviews the benefits and challenges associated with surface modification of the metals in biomedical applications. The paper also elaborates how the surface characteristics of metallic biomaterials, such as surface chemistry, topography, surface charge, and wettability, influence the protein adsorption and subsequent cell behavior in terms of adhesion, proliferation, and differentiation at the biomaterial–tissue interface. The chapter also highlights various techniques required for surface modification and coating of metallic biomaterials, including physicochemical and biochemical surface treatments and calcium phosphate and oxide coatings. In this review, the attention is focused on the biomaterial-associated infections, from which the need for anti-infective biomaterials originates. Biomaterial-associated infections differ markedly for epidemiology, aetiology and severity, depending mainly on the anatomic site, on the time of biomaterial application, and on the depth of the tissues harbouring the prosthesis. Here, the diversity and complexity of the different scenarios where medical devices are currently utilised are explored, providing an overview of the emblematic applicative fields and of the requirements for anti-infective biomaterials. In addition to this, chapter introduces nanomedicine and the use of both natural and synthetic polymeric biomaterials, focuses on specific current polymeric nanomedicine applications and research, and concludes with the challenges of nanomedicine research. Infection is currently regarded as the most severe and devastating complication associated to the use of biomaterials. Osteoporosis is a worldwide disease with a very high prevalence in humans older than 50. The main clinical consequences are bone fractures, which often lead to patient disability or even death. A number of commercial biomaterials are currently used to treat osteoporotic bone fractures, but most of these have not been specifically designed for that purpose. Many drug- or cell-loaded biomaterials have been proposed in research laboratories, but very few have received approval for commercial use. Polymeric nanomaterial-based therapeutics plays a key role in the field of medicine in treatment areas such as drug delivery, tissue engineering, cancer, diabetes, and neurodegenerative diseases. Advantages in the use of polymers over other materials for nanomedicine include increased functionality, design flexibility, improved processability, and, in some cases, biocompatibility.

Keywords: nanomedicine, tissue, infections, biomaterials

Procedia PDF Downloads 264
26 Accelerated Carbonation of Construction Materials by Using Slag from Steel and Metal Production as Substitute for Conventional Raw Materials

Authors: Karen Fuchs, Michael Prokein, Nils Mölders, Manfred Renner, Eckhard Weidner

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Due to the high CO₂ emissions, the energy consumption for the production of sand-lime bricks is of great concern. Especially the production of quicklime from limestone and the energy consumption for hydrothermal curing contribute to high CO₂ emissions. Hydrothermal curing is carried out under a saturated steam atmosphere at about 15 bar and 200°C for 12 hours. Therefore, we are investigating the opportunity to replace quicklime and sand in the production of building materials with different types of slag as calcium-rich waste from steel production. We are also investigating the possibility of substituting conventional hydrothermal curing with CO₂ curing. Six different slags (Linz-Donawitz (LD), ferrochrome (FeCr), ladle (LS), stainless steel (SS), ladle furnace (LF), electric arc furnace (EAF)) provided by "thyssenkrupp MillServices & Systems GmbH" were ground at "Loesche GmbH". Cylindrical blocks with a diameter of 100 mm were pressed at 12 MPa. The composition of the blocks varied between pure slag and mixtures of slag and sand. The effects of pressure, temperature, and time on the CO₂ curing process were studied in a 2-liter high-pressure autoclave. Pressures between 0.1 and 5 MPa, temperatures between 25 and 140°C, and curing times between 1 and 100 hours were considered. The quality of the CO₂-cured blocks was determined by measuring the compressive strength by "Ruhrbaustoffwerke GmbH & Co. KG." The degree of carbonation was determined by total inorganic carbon (TIC) and X-ray diffraction (XRD) measurements. The pH trends in the cross-section of the blocks were monitored using phenolphthalein as a liquid pH indicator. The parameter set that yielded the best performing material was tested on all slag types. In addition, the method was scaled to steel slag-based building blocks (240 mm x 115 mm x 60 mm) provided by "Ruhrbaustoffwerke GmbH & Co. KG" and CO₂-cured in a 20-liter high-pressure autoclave. The results show that CO₂ curing of building blocks consisting of pure wetted LD slag leads to severe cracking of the cylindrical specimens. The high CO₂ uptake leads to an expansion of the specimens. However, if LD slag is used only proportionally to replace quicklime completely and sand proportionally, dimensionally stable bricks with high compressive strength are produced. The tests to determine the optimum pressure and temperature show 2 MPa and 50°C as promising parameters for the CO₂ curing process. At these parameters and after 3 h, the compressive strength of LD slag blocks reaches the highest average value of almost 50 N/mm². This is more than double that of conventional sand-lime bricks. Longer CO₂ curing times do not result in higher compressive strengths. XRD and TIC measurements confirmed the formation of carbonates. All tested slag-based bricks show higher compressive strengths compared to conventional sand-lime bricks. However, the type of slag has a significant influence on the compressive strength values. The results of the tests in the 20-liter plant agreed well with the results of the 2-liter tests. With its comparatively moderate operating conditions, the CO₂ curing process has a high potential for saving CO₂ emissions.

Keywords: CO₂ curing, carbonation, CCU, steel slag

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25 All-In-One Universal Cartridge Based Truly Modular Electrolyte Analyzer

Authors: S. Dalvi, N. Sane, V. Patil, D. Bansode, A. Tharakan, V. Mathur

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Measurement of routine clinical electrolyte tests is common in labs worldwide for screening of illness or diseases. All the analyzers for the measurement of electrolyte parameters have sensors, reagents, sampler, pump tubing, valve, other tubing’s separate that are either expensive, require heavy maintenance and have a short shelf-life. Moreover, the costs required to maintain such Lab instrumentation is high and this limits the use of the device to only highly specialized personnel and sophisticated labs. In order to provide Healthcare Diagnostics to ALL at affordable costs, there is a need for an All-in-one Universal Modular Cartridge that contains sensors, reagents, sampler, valve, pump tubing, and other tubing’s in one single integrated module-in-module cartridge that is affordable, reliable, easy-to-use, requires very low sample volume and is truly modular and maintenance-free. DiaSys India has developed a World’s first, Patent Pending, Versatile All-in-one Universal Module-in-Module Cartridge based Electrolyte Analyzer (QDx InstaLyte) that can perform sodium, potassium, chloride, calcium, pH, lithium tests. QDx InstaLyte incorporates High Performance, Inexpensive All-in-one Universal Cartridge for rapid quantitative measurement of electrolytes in body fluids. Our proposed methodology utilizes Advanced & Improved long life ISE sensors to provide a sensitive and accurate result in 120 sec with just 100 µl of sample volume. The All-in-One Universal Cartridge has a very low reagent consumption capable of maximum of 1000 tests with a Use-life of 3-4 months and a long Shelf life of 12-18 months at 4-25°C making it very cost-effective. Methods: QDx InstaLyte analyzers with All-in-one Universal Modular Cartridges were independently evaluated with three R&D lots for Method Performance (Linearity, Precision, Method Comparison, Cartridge Stability) to measure Sodium, Potassium, Chloride. Method Comparison was done against Medica EasyLyte Plus Na/K/Cl Electrolyte Analyzer, a mid-size lab based clinical chemistry analyzer with N = 100 samples run over 10 days. Within-run precision study was done using modified CLSI guidelines with N = 20 samples and day-to-day precision study was done for 7 consecutive days using Trulab N & P Quality Control Samples. Accelerated stability testing was done at 45oC for 4 weeks with Production Lots. Results: Data analysis indicates that the CV for within-run precision for Na is ≤ 1%, for K is ≤2%, and for Cl is ≤2% and with R2 ≥ 0.95 for Method Comparison. Further, the All-in-One Universal Cartridge is stable up to 12-18 months at 4-25oC storage temperature based on preliminary extrapolated data. Conclusion: The Developed Technology Platform of All-in-One Universal Module-in-Module Cartridge based QDx InstaLyte is Reliable and meets all the performance specifications of the lab and is Truly Modular and Maintenance-Free. Hence, it can be easily adapted for low cost, sensitive and rapid measurement of electrolyte tests in low resource settings such as in urban, semi-urban and rural areas in the developing countries and can be used as a Point-of-care testing system for worldwide applications.

Keywords: all-in-one modular catridge, electrolytes, maintenance free, QDx instalyte

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24 Altered Proteostasis Contributes to Skeletal Muscle Atrophy during Chronic Hypobaric Hypoxia: An Insight into Signaling Mechanisms

Authors: Akanksha Agrawal, Richa Rathor, Geetha Suryakumar

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Muscle represents about ¾ of the body mass, and a healthy muscular system is required for human performance. A healthy muscular system is dynamically balanced via the catabolic and anabolic process. High altitude associated hypoxia altered this redox balance via producing reactive oxygen and nitrogen species that ultimately modulates protein structure and function, hence, disrupts proteostasis or protein homeostasis. The mechanism by which proteostasis is clinched includes regulated protein translation, protein folding, and protein degradation machinery. Perturbation in any of these mechanisms could increase proteome imbalance in the cellular processes. Altered proteostasis in skeletal muscle is likely to be responsible for contributing muscular atrophy in response to hypoxia. Therefore, we planned to elucidate the mechanism involving altered proteostasis leading to skeletal muscle atrophy under chronic hypobaric hypoxia. Material and Methods-Male Sprague Dawley rats weighing about 200-220 were divided into five groups - Control (Normoxic animals), 1d, 3d, 7d and 14d hypobaric hypoxia exposed animals. The animals were exposed to simulated hypoxia equivalent to 282 torr pressure (equivalent to an altitude of 7620m, 8% oxygen) at 25°C. On completion of chronic hypobaric hypoxia (CHH) exposure, rats were sacrificed, muscle was excised and biochemical, histopathological and protein synthesis signaling were studied. Results-A number of changes were observed with the CHH exposure time period. ROS was increased significantly on 07 and 14 days which were attributed to protein oxidation via damaging muscle protein structure by oxidation of amino acids moiety. The oxidative damage to the protein further enhanced the various protein degradation pathways. Calcium activated cysteine proteases and other intracellular proteases participate in protein turnover in muscles. Therefore, we analysed calpain and 20S proteosome activity which were noticeably increased at CHH exposure as compared to control group representing enhanced muscle protein catabolism. Since inflammatory markers (myokines) affect protein synthesis and triggers degradation machinery. So, we determined inflammatory pathway regulated under hypoxic environment. Other striking finding of the study was upregulation of Akt/PKB translational machinery that was increased on CHH exposure. Akt, p-Akt, p70 S6kinase, and GSK- 3β expression were upregulated till 7d of CHH exposure. Apoptosis related markers, caspase-3, caspase-9 and annexin V was also increased on CHH exposure. Conclusion: The present study provides evidence of disrupted proteostasis under chronic hypobaric hypoxia. A profound loss of muscle mass is accompanied by the muscle damage leading to apoptosis and cell death under CHH. These cellular stress response pathways may play a pivotal role in hypobaric hypoxia induced skeletal muscle atrophy. Further research in these signaling pathways will lead to development of therapeutic interventions for amelioration of hypoxia induced muscle atrophy.

Keywords: Akt/PKB translational machinery, chronic hypobaric hypoxia, muscle atrophy, protein degradation

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23 Pva-bg58s-cl-based Barrier Membranes For Guided Tissue/bone Regeneration Therapy

Authors: Isabela S. Gonçalves, Vitor G. P. Lima, Tiago M. B. Campos, Marcos Jacobovitz, Luana M. R. Vasconcellos, Ivone R. Oliveira

Abstract:

Periodontitis is an infectious disease of multifactorial origin, which originates from a periodontogenic bacterial biofilm that colonizes the surfaces of the teeth, resulting in an inflammatory reaction to microbial aggression. In the absence of adequate treatment, it can lead to the gradual destruction of the periodontal ligaments, cementum and alveolar bone. In guided tissue/bone regeneration therapy (GTR/GBR), a barrier membrane is placed between the fibrous tissues and the bone defect to prevent unwanted incursions of fibrous tissues into the bone defect, thus allowing the regeneration of quality bone. Currently, there are a significant number of biodegradable barrier membranes available on the market. However, a very common problem is that the membranes are not bioactive/osteogenic, that is, they are incapable of inducing a favorable osteogenic response and integration with the host tissue, resulting in many cases in displacement/expulsion of the membrane, requiring a new surgical procedure and replacement of the implant. Aiming to improve the bioactive and osteogenic properties of the membrane, this work evaluated the production of membranes that integrate the biocompatibility of the hydrophilic synthetic polymer (polyvinyl alcohol - PVA) with the osteogenic effects of chlorinated bioactive glasses (BG58S-Cl), using the electrospinning equipment (AeroSpinner L1.0 from Areka) which allows the execution of spinning by high voltage and/or blowing in solution and with a high production rate, enabling development on an industrial scale. In the formulation of bioactive glasses, the replacement of nitrates by chlorinated molecules has shown to be a promising alternative, since the chloride ion is naturally present in the body and, with its presence in the bioactive glass, the biocompatibility of the material increases. Thus, in this work, chlorinated bioactive glasses were synthesized by the sol-gel route using the compounds tetraethyl orthosilicate (TEOS), calcium chloride dihydrate and monobasic ammonium phosphate with pH adjustments with 37% HCl (1.5 or 2.5) and different calcination temperatures (500, 600 and 700 °C) were evaluated. The BG-58S-Cl powders obtained were characterized by pH, conductivity and zeta potential x time curves and by SEM/FEG, FTIR-ATR and Raman tests. The material produced under the selected conditions was evaluated in relation to the milling procedure, obtaining particles suitable for incorporation into PVA polymer solutions to be electrospun (D50 = 22 µm). Membranes were produced and evaluated regarding the influence of the crosslinking agent content as well as the crosslinking treatment temperature (3, 5 and 10 wt% citric acid) and (130 or 175 oC) and were characterized by SEM/FEG, FTIR, TG and DSC. From the optimization of the crosslinking conditions, membranes were prepared by adding BG58S-Cl powder (5 and 10 wt%) to the PVA solutions and were characterized by SEM-FEG, DSC, bioactivity in SBF and behavior in cell culture (cell viability, total protein content, alkaline phosphatase, mineralization nodules). The micrographs showed homogeneity of the distribution of BG58S-Cl particles throughout the sample, favoring cell differentiation.

Keywords: barrier membranes, chlorinated bioactive glasses, polyvinyl alcohol, tissue regeneration.

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22 Advanced Bio-Fuels for Biorefineries: Incorporation of Waste Tires and Calcium-Based Catalysts to the Pyrolysis of Biomass

Authors: Alberto Veses, Olga Sanhauja, María Soledad Callén, Tomás García

Abstract:

The appropriate use of renewable sources emerges as a decisive point to minimize the environmental impact caused by fossil fuels use. Particularly, the use of lignocellulosic biomass becomes one of the best promising alternatives since it is the only carbon-containing renewable source that can produce bioproducts similar to fossil fuels and it does not compete with food market. Among all the processes that can valorize lignocellulosic biomass, pyrolysis is an attractive alternative because it is the only thermochemical process that can produce a liquid biofuel (bio-oil) in a simple way and solid and gas fractions that can be used as energy sources to support the process. However, in order to incorporate bio-oils in current infrastructures and further process in future biorefineries, their quality needs to be improved. Introducing different low-cost catalysts and/or incorporating different polymer residues to the process are some of the new, simple and low-cost strategies that allow the user to directly obtain advanced bio-oils to be used in future biorefineries in an economic way. In this manner, from previous thermogravimetric analyses, local agricultural wastes such as grape seeds (GS) were selected as lignocellulosic biomass while, waste tires (WT) were selected as polymer residue. On the other hand, CaO was selected as low-cost catalyst based on previous experiences by the group. To reach this aim, a specially-designed fixed bed reactor using N₂ as a carrier gas was used. This reactor has the peculiarity to incorporate a vertical mobile liner that allows the user to introduce the feedstock in the oven once the selected temperature (550 ºC) is reached, ensuring higher heating rates needed for the process. Obtaining a well-defined phase distribution in the resulting bio-oil is crucial to ensure the viability to the process. Thus, once experiments were carried out, not only a well-defined two layers was observed introducing several mixtures (reaching values up to 40 wt.% of WT) but also, an upgraded organic phase, which is the one considered to be processed in further biorefineries. Radical interactions between GS and WT released during the pyrolysis process and dehydration reactions enhanced by CaO can promote the formation of better-quality bio-oils. The latter was reflected in a reduction of water and oxygen content of bio-oil and hence, a substantial increase of its heating value and its stability. Moreover, not only sulphur content was reduced from solely WT pyrolysis but also potential and negative issues related to a strong acidic environment of conventional bio-oils were minimized due to its basic pH and lower total acid numbers. Therefore, acidic compounds obtained in the pyrolysis such as CO₂-like substances can react with the CaO and minimize acidic problems related to lignocellulosic bio-oils. Moreover, this CO₂ capture promotes H₂ production from water gas shift reaction favoring hydrogen-transfer reactions, improving the final quality of the bio-oil. These results show the great potential of grapes seeds to carry out the catalytic co-pyrolysis process with different plastic residues in order to produce a liquid bio-oil that can be considered as a high-quality renewable vector.

Keywords: advanced bio-oils, biorefinery, catalytic co-pyrolysis of biomass and waste tires, lignocellulosic biomass

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21 Evaluation of Biological and Confinement Properties of a Bone Substitute to in Situ Preparation Based on Demineralized Bone Matrix for Bone Tissue Regeneration

Authors: Aura Maria Lopera Echavarria, Angela Maria Lema Perez, Daniela Medrano David, Pedronel Araque Marin, Marta Elena Londoño Lopez

Abstract:

Bone regeneration is the process by which the formation of new bone is stimulated. Bone fractures can originate at any time due to trauma, infections, tumors, congenital malformations or skeletal diseases. Currently there are different strategies to treat bone defects that in some cases, regeneration does not occur on its own. That is why they are treated with bone substitutes, which provide a necessary environment for the cells to synthesize new bone. The Demineralized Bone Matrix (DBM) is widely used as a bone implant due to its good properties, such as osteoinduction and bioactivity. However, the use of DBM is limited, because its presentation is powder, which is difficult to implant with precision and is susceptible to migrating to other sites through blood flow. That is why the DBM is commonly incorporated into a variety of vehicles or carriers. The objective of this project is to evaluate the bioactive and confinement properties of a bone substitute based on demineralized bone matrix (DBM). Also, structural and morphological properties were evaluated. Bone substitute was obtained from EIA Biomaterials Laboratory of EIA University and the DBM was facilitated by Tissue Bank Foundation. Morphological and structural properties were evaluated by scanning electron microscopy (SEM), X-ray diffraction (DRX) and Fourier transform infrared spectroscopy with total attenuated reflection (FTIR-ATR). Water absorption capacity and degradation were also evaluated during three months. The cytotoxicity was evaluated by the MTT test. The bioactivity of the bone substitute was evaluated through immersion of the samples in simulated body fluid during four weeks. Confinement tests were performed on tibial fragments of a human donor with bone defects of determined size, to ensure that the substitute remains in the defect despite the continuous flow of fluid. According of the knowledge of the authors, the methodology for evaluating samples in a confined environment has not been evaluated before in real human bones. The morphology of the samples showed irregular surface and presented some porosity. DRX confirmed a semi-crystalline structure. The FTIR-ATR determined the organic and inorganic phase of the sample. The degradation and absorption measurements stablished a loss of 3% and 150% in one month respectively. The MTT showed that the system is not cytotoxic. Apatite clusters formed from the first week were visualized by SEM and confirmed by EDS. These calcium phosphates are necessary to stimulate bone regeneration and thanks to the porosity of the developed material, osteinduction and osteoconduction are possible. The results of the in vitro evaluation of the confinement of the material showed that the migration of the bone filling to other sites is negligible, although the samples were subjected to the passage of simulated body fluid. The bone substitute, putty type, showed stability, is bioactive, non-cytotoxic and has handling properties for specialists at the time of implantation. The obtained system allows to maintain the osteoinductive properties of DBM and it can fill completely fractures in any way; however, it does not provide a structural support, that is, it should only be used to treat fractures without requiring a mechanical load.

Keywords: bone regeneration, cytotoxicity, demineralized bone matrix, hydrogel

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20 Index and Mechanical Geotechnical Properties and Their Control on the Strength and Durability of the Cainozoic Calcarenites in KwaZulu-Natal, South Africa

Authors: Luvuno N. Jele, Warwick W. Hastie, Andrew Green

Abstract:

Calcarenite is a clastic sedimentary beach rock composed of more than 50% sand sized (0.0625 – 2 mm) carbonate grains. In South Africa, these rocks occur as a narrow belt along most of the coast of KwaZulu-Natal and sporadically along the coast of the Eastern Cape. Calcarenites contain a high percentage of calcium carbonate, and due to a number of its physical and structural features, like porosity, cementing material, sedimentary structures, grain shape, and grain size; they are more prone to chemical and mechanical weathering. The objective of the research is to study the strength and compressibility characteristics of the calcarenites along the coast of KwaZulu-Natal to be able to better understand the geotechnical behaviour of these rocks, which may help to predict areas along the coast which may be potentially susceptible to failure/differential settling resulting in damage to property. A total of 148 cores were prepared and analyzed. Cores were analyzed perpendicular and parallel to bedding. Tests were carried out in accordance with the relevant codes and recommendations of the International Society for Rock Mechanics, American Standard Testing Methods, and Committee of Land and Transport Standard Specifications for Road and Bridge Works for State Road Authorities. Test carried out included: x-ray diffraction, petrography, shape preferred orientation (SPO), 3-D Tomography, rock porosity, rock permeability, ethylene glycol, slake durability, rock water absorption, Duncan swelling index, triaxial compressive strength, Brazilian tensile strength and uniaxial compression test with elastic modulus. The beach-rocks have a uniaxial compressive strength (UCS) ranging from 17,84Mpa to 287,35Mpa and exhibit three types of failure; (1) single sliding shear failure, (2) complete cone development, and (3) splitting failure. Brazilian tensile strength of the rocks ranges from 2.56 Mpa to 12,40 Ma, with those tested perpendicular to bedding showing lower tensile strength. Triaxial compressive tests indicate calcarenites have strength ranging from 86,10 Mpa to 371,85 Mpa. Common failure mode in the triaxial test is a single sliding shear failure. Porosity of the rocks varies from 1.25 % to 26.52 %. Rock tests indicate that the direction of loading, whether it be parallel to bedding or perpendicular to bedding, plays no significantrole in the strength and durability of the calcarenites. Porosity, cement type, and grain texture play major roles.UCS results indicate that saturated cores are weaker in strength compared to dry samples. Thus, water or moisture content plays a significant role in the strength and durability of the beach-rock. Loosely packed, highly porous and low magnesian-calcite bearing calcarenites show a decrease in strength compared to the densely packed, low porosity and high magnesian-calcite bearing calcarenites.

Keywords: beach-rock, calcarenite, cement, compressive, failure, porosity, strength, tensile, grains

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19 Testicular Differential MicroRNA Expression Derived Occupational Risk Factor Assessment in Idiopathic Non-obstructive Azoospermia Cases

Authors: Nisha Sharma, Mili Kaur, Ashutosh Halder, Seema Kaushal, Manoj Kumar, Manish Jain

Abstract:

Purpose: To investigate microRNAs (miRNA) as an epigenomic etiological factor in idiopathic non-obstructive azoospermia (NOA). In order to achieve the same, an association was seen between occupational exposure to radiation, thermal, and chemical factors and idiopathic cases of non-obstructive azoospermia, and later, testicular differential miRNA expression profiling was done in exposure group NOA cases. Method: It is a prospective study in which 200 apparent idiopathic male factor infertility cases, who have been advised to undergo testicular fine needle aspiration (FNA) evaluation, are recruited. A detailed occupational history was taken to understand the possible type of exposure due to the nature and duration of work. A total of 26 patients were excluded upon XY-FISH and Yq microdeletion tests due to the presence of genetic causes of infertility, 6 hypospermatogeneis (HS), six Sertoli cell-only syndrome (SCOS), and six normospermatogeneis patients testicular FNA samples were used for RNA isolation followed by small RNA sequencing and nCounter miRNA expression analysis. Differential miRNA expression profile of HS and SCOS patients was done. A web-based tool, miRNet, was used to predict the interacting compounds or chemicals using the shortlisted miRNAs with high fold change. The major limitation encountered in this study was the insufficient quantity of testicular FNA sample used for total RNA isolation, which resulted in a low yield and RNA integrity number (RIN) value. Therefore, the number of RNA samples admissible for differential miRNA expression analysis was very small in comparison to the total number of patients recruited. Results: Differential expression analysis revealed 69 down-regulated and 40 up-regulated miRNAs in HS and 66 down-regulated and 33 up-regulated miRNAs in SCOS in comparison to normospermatogenesis controls. The miRNA interaction analysis using the miRNet tool showed that the differential expression profiles of HS and SCOS patients were associated with arsenic trioxide, bisphenol-A, calcium sulphate, lithium, and cadmium. These compounds are reproductive toxins and might be responsible for miRNA-mediated epigenetic deregulation leading to NOA. The association between occupational risk factor exposure and the non-exposure group of NOA patients was not statistically significant, with ꭓ2 (3, N= 178) = 6.70, p= 0.082. The association between individual exposure groups (radiation, thermal, and chemical) and various sub-types of NOA is also not significant, with ꭓ2 (9, N= 178) = 15.06, p= 0.089. Functional analysis of HS and SCOS patients' miRNA profiles revealed some important miR-family members in terms of male fertility. The miR-181 family plays a role in the differentiation of spermatogonia and spermatocytes, as well as the transcriptional regulation of haploid germ cells. The miR-34 family is expressed in spermatocytes and round spermatids and is involved in the regulation of SSCs differentiation. Conclusion: The reproductive toxins might adopt the miRNA-mediated mechanism of disease development in idiopathic cases of NOA. Chemical compound induced; miRNA-mediated epigenetic deregulation can give a future perspective on the etiopathogenesis of the disease.

Keywords: microRNA, non-obstructive azoospermia (NOA), occupational exposure, hypospermatogenesis (HS), Sertoli cell only syndrome (SCOS)

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18 Genome-Wide Analysis Identifies Locus Associated with Parathyroid Hormone Levels

Authors: Antonela Matana, Dubravka Brdar, Vesela Torlak, Marijana Popovic, Ivana Gunjaca, Ozren Polasek, Vesna Boraska Perica, Maja Barbalic, Ante Punda, Caroline Hayward, Tatijana Zemunik

Abstract:

Parathyroid hormone (PTH) plays a critical role in the regulation of bone mineral metabolism and calcium homeostasis. Higher PTH levels are associated with heart failure, hypertension, coronary artery disease, cardiovascular mortality and poorer bone health. A twin study estimated that 60% of the variation in PTH concentrations is genetically determined. Only one GWAS of PTH concentration has been reported to date. Identified loci explained 4.5% of the variance in circulating PTH, suggesting that additional genetic variants remain undiscovered. Therefore, the aim of this study was to identify novel genetic variants associated with PTH levels in a general population. We have performed a GWAS meta-analysis on 2596 individuals originating from three Croatian cohorts: City of Split and the Islands of Korčula and Vis, within a large-scale project of “10,001 Dalmatians”. A total of 7 411 206 variants, imputed using the 1000 Genomes reference panel, with minor allele frequency ≥ 1% and Rsq ≥ 0.5 were analyzed for the association. GWAS within each data set was performed under an additive model, controlling for age, gender and relatedness. Meta-analysis was conducted using the inverse-variance fixed-effects method. Furthermore, to identify sex-specific effects, we have conducted GWAS meta-analyses analyzing males and females separately. In addition, we have performed biological pathway analysis. Four SNPs, representing one locus, reached genome-wide significance. The most significant SNP was rs11099476 on chromosome 4 (P=1.15x10-8), which explained 1.14 % of the variance in PTH. The SNP is located near the protein-coding gene RASGEF1B. Additionally, we detected suggestive association with SNPs, rs77178854 located on chromosome 2 in the DPP10 gene (P=2.46x10-7) and rs481121 located on chromosome 1 (P=3.58x10-7) near the GRIK1 gene. One of the top hits detected in the main meta-analysis, intron variant rs77178854 located within DPP10 gene, reached genome-wide significance in females (P=2.21x10-9). No single locus was identified in the meta-analysis in males. Fifteen biological pathways were functionally enriched at a P<0.01, including muscle contraction, ion homeostasis and cardiac conduction as the most significant pathways. RASGEF1B is the guanine nucleotide exchange factor, known to be associated with height, bone density, and hip. DPP10 encodes a membrane protein that is a member of the serine proteases family, which binds specific voltage-gated potassium channels and alters their expression and biophysical properties. In conclusion, we identified 2 novel loci associated with PTH levels in a general population, providing us with further insights into the genetics of this complex trait.

Keywords: general population, genome-wide association analysis, parathyroid hormone, single nucleotide polymorphisms.

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17 The 10,000 Fold Effect of Retrograde Neurotransmission, a New Concept for Stroke Revival: Use of Intracarotid Sodium Nitroprusside

Authors: Vinod Kumar

Abstract:

Background: Tissue Plasminogen Activator (tPA) showed a level 1 benefit in acute stroke (within 3-6 hrs). Intracarotid sodium nitroprusside (ICSNP) has been studied in this context with a wide treatment window, fast recovery and affordability. This work proposes two mechanisms for acute cases and one mechanism for chronic cases, which are interrelated, for physiological recovery. a)Retrograde Neurotransmission (acute cases): 1)Normal excitatory impulse: at the synaptic level, glutamate activates NMDA receptors, with nitric oxide synthetase (NOS) on the postsynaptic membrane, for further propagation by the calcium-calmodulin complex. Nitric oxide (NO, produced by NOS) travels backward across the chemical synapse and binds the axon-terminal NO receptor/sGC of a presynaptic neuron, regulating anterograde neurotransmission (ANT) via retrograde neurotransmission (RNT). Heme is the ligand-binding site of the NO receptor/sGC. Heme exhibits > 10,000-fold higher affinity for NO than for oxygen (the 10,000-fold effect) and is completed in 20 msec. 2)Pathological conditions: normal synaptic activity, including both ANT and RNT, is absent. A NO donor (SNP) releases NO from NOS in the postsynaptic region. NO travels backward across a chemical synapse to bind to the heme of a NO receptor in the axon terminal of a presynaptic neuron, generating an impulse, as under normal conditions. b)Vasospasm: (acute cases) Perforators show vasospastic activity. NO vasodilates the perforators via the NO-cAMP pathway. c)Long-Term Potentıatıon (LTP): (chronic cases) The NO–cGMP-pathway plays a role in LTP at many synapses throughout the CNS and at the neuromuscular junction. LTP has been reviewed both generally and with respect to brain regions specific for memory/learning. Aims/Study Des’gn: The principles of “generation of impulses from the presynaptic region to the postsynaptic region by very potent RNT (10,000-fold effect)” and “vasodilation of arteriolar perforators” are the basis of the authors’ hypothesis to treat stroke cases. Case-control prospective study. Mater’als And Methods: The experimental population included 82 stroke patients (10 patients were given control treatments without superfusion or with 5% dextrose superfusion, and 72 patients comprised the ICSNP group). The mean time for superfusion was 9.5 days post-stroke. Pre- and post-ICSNP status was monitored by NIHSS, MRI and TCD. Results: After 90 seconds in the ICSNP group, the mean change in the NIHSS score was a decrease of 1.44 points, or 6.55%; after 2 h, there was a decrease of 1.16 points; after 24 h, there was an increase of 0.66 points, 2.25%, compared to the control-group increase of 0.7 points, or 3.53%; at 7 days, there was an 8.61-point decrease, 44.58%, compared to the control-group increase of 2.55 points, or 22.37%; at 2 months in ICSNP, there was a 6.94-points decrease, 62.80%, compared to the control-group decrease of 2.77 points, or 8.78%. TCD was documented and improvements were noted. Conclusions: ICSNP is a swift-acting drug in the treatment of stroke, acting within 90 seconds on day 9.5 post-stroke with a small decrease after 24 hours. The drug recovers from this decrease quickly.

Keywords: brain infarcts, intracarotid sodium nitroprusside, perforators, vasodilatıons, retrograde transmission, the 10, 000-fold effect

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16 Structural Molecular Dynamics Modelling of FH2 Domain of Formin DAAM

Authors: Rauan Sakenov, Peter Bukovics, Peter Gaszler, Veronika Tokacs-Kollar, Beata Bugyi

Abstract:

FH2 (formin homology-2) domains of several proteins, collectively known as formins, including DAAM, DAAM1 and mDia1, promote G-actin nucleation and elongation. FH2 domains of these formins exist as oligomers. Chain dimerization by ring structure formation serves as a structural basis for actin polymerization function of FH2 domain. Proper single chain configuration and specific interactions between its various regions are necessary for individual chains to form a dimer functional in G-actin nucleation and elongation. FH1 and WH2 domain-containing formins were shown to behave as intrinsically disordered proteins. Thus, the aim of this research was to study structural dynamics of FH2 domain of DAAM. To investigate structural features of FH2 domain of DAAM, molecular dynamics simulation of chain A of FH2 domain of DAAM solvated in water box in 50 mM NaCl was conducted at temperatures from 293.15 to 353.15K, with VMD 1.9.2, NAMD 2.14 and Amber Tools 21 using 2z6e and 1v9d PDB structures of DAAM was obtained on I-TASSER webserver. Calcium and ATP bound G-actin 3hbt PDB structure was used as a reference protein with well-described structural dynamics of denaturation. Topology and parameter information of CHARMM 2012 additive all-atom force fields for proteins, carbohydrate derivatives, water and ions were used in NAMD 2.14 and ff19SB force field for proteins in Amber Tools 21. The systems were energy minimized for the first 1000 steps, equilibrated and produced in NPT ensemble for 1ns using stochastic Langevin dynamics and the particle mesh Ewald method. Our root-mean square deviation (RMSD) analysis of molecular dynamics of chain A of FH2 domains of DAAM revealed similar insignificant changes of total molecular average RMSD values of FH2 domain of these formins at temperatures from 293.15 to 353.15K. In contrast, total molecular average RMSD values of G-actin showed considerable increase at 328K, which corresponds to the denaturation of G-actin molecule at this temperature and its transition from native, ordered, to denatured, disordered, state which is well-described in the literature. RMSD values of lasso and tail regions of chain A of FH2 domain of DAAM exhibited higher than total molecular average RMSD at temperatures from 293.15 to 353.15K. These regions are functional in intra- and interchain interactions and contain highly conserved tryptophan residues of lasso region, highly conserved GNYMN sequence of post region and amino acids of the shell of hydrophobic pocket of the salt bridge between Arg171 and Asp321, which are important for structural stability and ordered state of FH2 domain of DAAM and its functions in FH2 domain dimerization. In conclusion, higher than total molecular average RMSD values of lasso and post regions of chain A of FH2 domain of DAAM may explain disordered state of FH2 domain of DAAM at temperatures from 293.15 to 353.15K. Finally, absence of marked transition, in terms of significant changes in average molecular RMSD values between native and denatured states of FH2 domain of DAAM at temperatures from 293.15 to 353.15K, can make it possible to attribute these formins to the group of intrinsically disordered proteins rather than to the group of intrinsically ordered proteins such as G-actin.

Keywords: FH2 domain, DAAM, formins, molecular modelling, computational biophysics

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15 The 10,000 Fold Effect of Retrograde Neurotransmission: A New Concept for Cerebral Palsy Revival by the Use of Nitric Oxide Donars

Authors: V. K. Tewari, M. Hussain, H. K. D. Gupta

Abstract:

Background: Nitric Oxide Donars (NODs) (intrathecal sodium nitroprusside (ITSNP) and oral tadalafil 20mg post ITSNP) has been studied in this context in cerebral palsy patients for fast recovery. This work proposes two mechanisms for acute cases and one mechanism for chronic cases, which are interrelated, for physiological recovery. a) Retrograde Neurotransmission (acute cases): 1) Normal excitatory impulse: at the synaptic level, glutamate activates NMDA receptors, with nitric oxide synthetase (NOS) on the postsynaptic membrane, for further propagation by the calcium-calmodulin complex. Nitric oxide (NO, produced by NOS) travels backward across the chemical synapse and binds the axon-terminal NO receptor/sGC of a presynaptic neuron, regulating anterograde neurotransmission (ANT) via retrograde neurotransmission (RNT). Heme is the ligand-binding site of the NO receptor/sGC. Heme exhibits > 10,000-fold higher affinity for NO than for oxygen (the 10,000-fold effect) and is completed in 20 msec. 2) Pathological conditions: normal synaptic activity, including both ANT and RNT, is absent. A NO donor (SNP) releases NO from NOS in the postsynaptic region. NO travels backward across a chemical synapse to bind to the heme of a NO receptor in the axon terminal of a presynaptic neuron, generating an impulse, as under normal conditions. b) Vasopasm: (acute cases) Perforators show vasospastic activity. NO vasodilates the perforators via the NO-cAMP pathway. c) Long-Term Potentiation (LTP): (chronic cases) The NO–cGMP-pathway plays a role in LTP at many synapses throughout the CNS and at the neuromuscular junction. LTP has been reviewed both generally and with respect to brain regions specific for memory/learning. Aims/Study Design: The principles of “generation of impulses from the presynaptic region to the postsynaptic region by very potent RNT (10,000-fold effect)” and “vasodilation of arteriolar perforators” are the basis of the authors’ hypothesis to treat cerebral palsy cases. Case-control prospective study. Materials and Methods: The experimental population included 82 cerebral palsy patients (10 patients were given control treatments without NOD or with 5% dextrose superfusion, and 72 patients comprised the NOD group). The mean time for superfusion was 5 months post-cerebral palsy. Pre- and post-NOD status was monitored by Gross Motor Function Classification System for Cerebral Palsy (GMFCS), MRI, and TCD studies. Results: After 7 days in the NOD group, the mean change in the GMFCS score was an increase of 1.2 points mean; after 3 months, there was an increase of 3.4 points mean, compared to the control-group increase of 0.1 points at 3 months. MRI and TCD documented the improvements. Conclusions: NOD (ITSNP boosts up the recovery and oral tadalafil maintains the recovery to a well-desired level) acts swiftly in the treatment of CP, acting within 7 days on 5 months post-cerebral palsy either of the three mechanisms.

Keywords: cerebral palsy, intrathecal sodium nitroprusside, oral tadalafil, perforators, vasodilations, retrograde transmission, the 10, 000-fold effect, long-term potantiation

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14 Regulation Effect of Intestinal Microbiota by Fermented Processing Wastewater of Yuba

Authors: Ting Wu, Feiting Hu, Xinyue Zhang, Shuxin Tang, Xiaoyun Xu

Abstract:

As a by-product of yuba, processing wastewater of Yuba (PWY) contains many bioactive components such as soybean isoflavones, soybean polysaccharides and soybean oligosaccharides, which is a good source of prebiotics and has a potential of high value utilization. The use of Lactobacillus plantarum to ferment PWY can be considered as a potential biogenic element, which can regulate the balance of intestinal microbiota. In this study, firstly, Lactobacillus plantarum was used to ferment PWY to improve its content of active components and antioxidant activity. Then, the health effect of fermented processing wastewater of yuba (FPWY) was measured in vitro. Finally, microencapsulation technology was used applied to improve the sustained release of FPWY and reduce the loss of active components in the digestion process, as well as to improving the activity of FPWY. The main results are as follows: (1) FPWY presented a good antioxidant capacity with DPPH free radical scavenging ability (0.83 ± 0.01 mmol Trolox/L), ABTS free radical scavenging ability (7.47 ± 0.35 mmol Trolox/L) and iron ion reducing ability (1.11 ± 0.07 mmol Trolox/L). Compared with non-fermented processing wastewater of yuba (NFPWY), there was no significant difference in the content of total soybean isoflavones, but the content of glucoside soybean isoflavones decreased, and aglyconic soybean isoflavones increased significantly. After fermentation, PWY can effectively reduce the soluble monosaccharides, disaccharides and oligosaccharides, such as glucose, fructose, galactose, trehalose, stachyose, maltose, raffinose and sucrose. (2) FPWY can significantly enhance the growth of beneficial bacteria such as Bifidobacterium, Ruminococcus and Akkermansia, significantly inhibit the growth of harmful bacteria E.coli, regulate the structure of intestinal microbiota, and significantly increase the content of short-chain fatty acids such as acetic acid, propionic acid, butyric acid, isovaleric acid. Higher amount of lactic acid in the gut can be further broken down into short chain fatty acids. (3) In order to improve the stability of soybean isoflavones in FPWY during digestion, sodium alginate and chitosan were used as wall materials for embedding. The FPWY freeze-dried powder was embedded by the method of acute-coagulation bath. The results show that when the core wall ratio is 3:1, the concentration of chitosan is 1.5%, the concentration of sodium alginate is 2.0%, and the concentration of calcium is 3%, the embossing rate is 53.20%. In the simulated in vitro digestion stage, the release rate of microcapsules reached 59.36% at the end of gastric digestion and 82.90% at the end of intestinal digestion. Therefore, the core materials with good sustained-release performance of microcapsules were almost all released. The structural analysis results of FPWY microcapsules show that the microcapsules have good mechanical properties. Its hardness, springness, cohesiveness, gumminess, chewiness and resilience were 117.75± 0.21 g, 0.76±0.02, 0.54±0.01, 63.28±0.71 g·sec, 48.03±1.37 g·sec, 0.31±0.01, respectively. Compared with the unembedded FPWY, the infrared spectrum results showed that the microcapsules had embedded effect on the FPWY freeze-dried powder.

Keywords: processing wastewater of yuba, lactobacillus plantarum, intestinal microbiota, microcapsule

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13 The Use of STIMULAN Resorbable Antibiotic Beads in Conjunction with Autologous Tissue Transfer to Treat Recalcitrant Infections and Osteomyelitis in Diabetic Foot Wounds

Authors: Hayden R Schott, John M Felder III

Abstract:

Introduction: Chronic lower extremity wounds in the diabetic and vasculopathic populations are associated with a high degree of morbidity.When wounds require more extensive treatment than can be offered by wound care centers, more aggressive solutions involve local tissue transfer and microsurgical free tissue transfer for achieving definitive soft tissue coverage. These procedures of autologous tissue transfer (ATT) offer resilient, soft tissue coverage of limb-threatening wounds and confer promising limb salvage rates. However, chronic osteomyelitis and recalcitrant soft tissue infections are common in severe diabetic foot wounds and serve to significantly complicate ATT procedures. Stimulan is a resorbable calcium sulfate antibiotic carrier. The use of stimulan antibiotic beads to treat chronic osteomyelitis is well established in the orthopedic and plastic surgery literature. In these procedures, the beads are placed beneath the skin flap to directly deliver antibiotics to the infection site. The purpose of this study was to quantify the success of Stimulan antibiotic beads in treating recalcitrant infections in patients with diabetic foot wounds receiving ATT. Methods: A retrospective review of clinical and demographic information was performed on patients who underwent ATT with the placement of Stimulan antibiotic beads for attempted limb salvage from 2018-21. Patients were analyzed for preoperative wound characteristics, demographics, infection recurrence, and adverse outcomes as a result of product use. The primary endpoint was 90 day infection recurrence, with secondary endpoints including 90 day complications. Outcomes were compared using basic statistics and Fisher’s exact tests. Results: In this time span, 14 patients were identified. At the time of surgery, all patients exhibited clinical signs of active infection, including positive cultures and erythema. 57% of patients (n=8) exhibited chronic osteomyelitis prior to surgery, and 71% (n=10) had exposed bone at the wound base. In 57% of patients (n=8), Stimulan beads were placed beneath a free tissue flap and beneath a pedicle tissue flap in 42% of patients (n=6). In all patients, Stimulan beads were only applied once. Recurrent infections were observed in 28% of patients (n=4) at 90 days post-op, and flap nonadherence was observed in 7% (n=1). These were the only Stimulan related complications observed. Ultimately, lower limb salvage was successful in 85% of patients (n=12). Notably, there was no significant association between the preoperative presence of osteomyelitis and recurrent infections. Conclusions: The use of Stimulanantiobiotic beads to treat recalcitrant infections in patients receiving definitive skin coverage of diabetic foot wounds does not appear to demonstrate unnecessary risk. Furthermore, the lack of significance between the preoperative presence of osteomyelitis and recurrent infections indicates the successful use of Stimulan to dampen infection in patients with osteomyelitis, as is consistent with the literature. Further research is needed to identify Stimulan as the significant contributor to infection treatment using future cohort and case control studies with more patients. Nonetheless, the use of Stimulan antibiotic beads in patients with diabetic foot wounds demonstrates successful infection suppression and maintenance of definitive soft tissue coverage.

Keywords: wound care, stimulan antibiotic beads, free tissue transfer, plastic surgery, wound, infection

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