Search results for: phosphate buffer

Authors: Rym Saidi, Pape Alioune Ndiaye, Ibnyasser Ammar, Zineb Rchiad, Khalid Daoui, Issam Kadmiri Meftahi, Adnane Bargaz

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Chickpea (Cicer arietinum L.) is an important leguminous crop grown worldwide and plays a significant role in humans’ dietary consumption. Alongside nitrogen (N), low phosphorus (P) availability within agricultural soils is one of the major factors limiting chickpea growth and productivity. The combined application of beneficial bacterial inoculants and Rock P-fertilizer could boost chickpea performance and productivity, increasing P-utilization efficiency and minimizing nutrient losses under P-deficiency conditions. A greenhouse experiment was conducted to evaluate the response of chickpeas to two P-fertilizer forms (RP and TSP) under N2-fixer and P-solubilizer consortium inoculation to improve biological N fixation and P nutrition under P-deficient conditions. Under inoculation, chickpea chlorophyll content and chlorophyll fluorescence (RP+I and TSP+I) were increased compared to uninoculated treatments. The RP+I treatment increased both shoot and root dry weights by 48,80% and 72,68%, respectively, compared to the uninoculated RP fertilized control. Indeed, the bacterial consortium contributed to enhancing root morphological traits (e.g., root volume, surface area, and diameter) of all inoculated treatments versus the uninoculated treatments. Furthermore, soil available P and root inorganic P were significantly improved in RP+I by 162,84% and 73,24%, respectively, compared to uninoculated RP control. Our research outcomes suggest that the co-inoculation of chickpeas with N2-fixing, and P-solubilizing bacteria improves biomass yield and nutrient uptake. Eventually, enhancing chickpea agrophysiological performance, especially in restricted P-availability conditions.

Keywords: chickpea, consortium, beneficial bacterial inoculants, phosphorus deficiency, rock p-fertilizer, nutrient uptake

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Authors: G. Balaji, R. Balasundaraprabhu, S. Prasanna, M. D. Kannan, K. Sivakumaran, David Mcilroy

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Copper Zin tin sulphide (Cu2ZnSnS4 or CZTS) is found to be better alternative to Copper Indium gallium diselenide as absorber layers in thin film based solar cells due to the utilisation of earth-abundant materials in the midst of lower toxicity. In the present study, Cu2ZnSnS4 thin films were prepared on soda lime glass using (CuS, ZnS, SnS) targets and were deposited by three different stacking orders, using RF Magnetron sputtering. The substrate temperature was fixed at 300 °C during the depositions. CZTS thin films were characterized using X-ray diffraction, X-ray photoelectron spectroscopy, Raman spectroscopy and UV-Vis-NIR spectroscopy. All the samples exhibited X-ray peaks pertaining to (112) kesterite phase of CZTS, along with the presence of a predominant wurtzite CZTS phase. X-ray photoelectron spectroscopy revealed the presence of all the elements in all the samples. The change in stacking order clearly shows that it affects the structural and phase properties of the films. Relative atomic concentrations of Zn, Cu, Sn and S, which are determined by high-resolution XPS core level spectra integrated peak areas revealed that the CZTS films exhibit inhomogeneity in both stoichiometry and elemental composition. Raman spectroscopy studies on the film showed the presence of CZTS phase. The energy band gap of the CZTS thin films was found to be in the range of 1.5 eV to 1.6 eV. The films were then annealed at 450 °C for 5 hrs and it was found that the predominant nature of the X-ray peaks has transformed from Wurtzite to Kesterite phase which is highly desirable for absorber layers in thin film solar cells. The optimized CZTS layer was used as an absorber layer in thin film solar cells. ZnS and CdS were used as buffer layers which in turn prepared by Hot wall epitaxy technique. Gallium doped Zinc oxide was used as a transparent conducting oxide. The solar cell structure Glass/Mo/CZTS/CdS or ZnS/GZO has been fabricated, and solar cell parameters were measured.

Keywords: earth-abundant, Kesterite, RF sputtering, thin film solar cells

Procedia PDF Downloads 264

Authors: Yi-Shuan ShaoShao, Yen-An Tsai, Chia-Huang Chang, Kai-Wei Liao, Ming-Song Tsai, Mei-Lien Chen

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Organophosphate pesticides (OPs) is an environmental hormone with proven endocrine-disrupting effects that may affect the growth and development in human. A large amount of organophosphate pesticides (OPs) is used throughout Taiwan, and human may be exposed through dietary intake or residential use. During pregnancy, OPs can be transferred to the blood stream reaching the fetus through the placenta. The aim of this study was to explore the association between maternal OPs exposure levels and fetal developments and birth outcomes. A birth cohort was follow-up. Maternal urine sample were collected at the first, second, and third gestational trimester. Fetal growth characteristics were measured by ultrasonic scan and birth outcomes were assessed by pediatrician. Urinary metabolite of organophosphate pesticides were assessed using gas chromatography-mass spectrometry. The analytes included dimethylphosphate (DMP), dimethylthiophosphate (DMTP), dimethyldithiophosphates (DMDTP), diethylphosphate (DEP), diethylthiophosphate (DETP), and diethyldithiophosphate (DEDTP). We found that all of urine samples in each trimester were detected at least one metabolite for dialkyl phosphate (DAP). The detection rate range of OP urinary metabolites were from the lowest 22% DEDTP to the highest 100% DMP and DMTP. And to compared geometric means (GM) of urinary metabolites with three trimesters, that third trimester had the highest concentration for DMPs, DEPs, and DAPs in pregnant women were 368.01, 169.85 and 543.75 nmol/g creatinine, respectively. We observed that DAPs concentration in first and second trimester were significantly negative association with head circumference. DMPs in first trimester was significantly negative association with thoracic circumference (p=0.05) by spearman correlation. Our results support associations with prenatal OPs exposure with fetal head circumference and thoracic circumference. It provided that maternal OPs exposure might affect birth outcomes. Thus, prenatal exposure to OPs and health risk worthy of attention and concern.

Keywords: DAPs, birth outcomes, organophosphate pesticides, prenatal

Procedia PDF Downloads 317

Authors: Rajeev Kumar, Angad Singh Kushwaha, Amritanshu Pandey, S. K. Srivastava

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Photonic crystals (PCs) have attracted much attention due to its electromagnetic properties and potential applications. In PCs, there is certain range of wavelength where electromagnetic waves are not allowed to pass are called photonic band gap (PBG). A localized defect mode will appear within PBG, due to change in the interference behavior of light, when we create a defect in the periodic structure. We can also create different types of defect structures by inserting or removing a layer from the periodic layered structure in two and three-dimensional PCs. We can design microcavity, waveguide, and perfect mirror by creating a point defect, line defect, and palanar defect in two and three- dimensional PC structure. One-dimensional and two-dimensional PCs with defects were reported theoretically and experimentally by Smith et al.. in conventional photonic band gap structure. In the present paper, we have presented the defect mode tunability in tilted non-graded photonic crystal (NGPC) and linearly graded photonic crystal (LGPC) using lead sulphide (PbS) and titanium dioxide (TiO2) in the infrared region. A birefringent defect layer is created in NGPC and LGPC using potassium titany phosphate (KTP). With the help of transfer matrix method, the transmission properties of proposed structure is investigated for transverse electric (TE) and transverse magnetic (TM) polarization. NGPC and LGPC without defect layer is also investigated. We have found that a photonic band gap (PBG) arises in the infrared region. An additional defect layer of KTP is created in NGPC and LGPC structure. We have seen that an additional transmission mode appers in PBG region. It is due to the addition of defect layer. We have also seen the effect, linear gradation in thickness, angle of incidence, tilt angle, and thickness of defect layer, on PBG and additional transmission mode. We have observed that the additional transmission mode and PBG can be tuned by changing the above parameters. The proposed structure may be used as channeled filter, optical switches, monochromator, and broadband optical reflector.

Keywords: defect modes, graded photonic crystal, photonic crystal, tilt angle

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Authors: Huang Bingling, Jia Yuhong, Liu Yanhui

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As an important part of aircraft, landing gears are responsible for taking-off and landing function. In recent years, big airplane's structural quality increases a lot. As a result, landing gears have stricter technical requirements than ever before such as structure strength and etc. If the structural strength of the landing gear is enhanced through traditional methods like increasing structural quality, the negative impacts on the landing gear's function would be very serious and even counteract the positive effects. Thus, in order to solve this problem, the impact of pre-rotating of landing gears on performance of landing gears is studied from the theoretical and experimental verification in this paper. By increasing the pre-rotating speed of the wheel, it can improve the performance of the landing gear and reduce the structural quality, the force of joint parts and other properties. In addition, the pre-rotating of the wheels also has other advantages, such as reduce the friction between wheels and ground and extend the life of the wheel. In this paper, the impact of the pre-rotating speed on landing gears and the connecting between landing gears performance and pre-rotating speed would be researched in detail. This paper is divided into three parts. In the first part, large airplane landing gear model is built by CATIA and LMS. As most general landing gear type in big plane, four-wheel landing gear is picked as model. The second part is to simulate the process of landing in LMS motion, and study the impact of pre-rotating of wheels on the aircraft`s properties, including the buffer stroke, efficiency, power; friction, displacement and relative speed between piston and sleeve; force and load distribution of tires. The simulation results show that the characteristics of the different pre-rotation speed are understood. The third part is conclusion. Through the data of the previous simulation and the relationship between the pre-rotation speed of the aircraft wheels and the performance of the aircraft, recommended speed interval is proposed. This paper is of great theoretical value to improve the performance of large airplane. It is a very effective method to improve the performance of aircraft by setting wheel pre-rotating speed. Do not need to increase the structural quality too much, eliminating the negative effects of traditional methods.

Keywords: large airplane, landing gear, pre-rotating, simulation

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Authors: Prateek Singh, Dilshad Ahmad

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Growing demand for high-quality steel products has inspired researchers to investigate the unit operations involved in the manufacturing of these products (slabs, rods, sheets, etc.). One such operation is tundish operation, in which a vessel (tundish) acts as a buffer of molten steel for the solidification operation in mold. It is observed that tundish also plays a crucial role in the quality and cleanliness of the steel produced, besides merely acting as a reservoir for the mold. It facilitates removal of dissolved oxygen (inclusions) from the molten steel thus improving its cleanliness. Inclusion removal can be enhanced by increasing the residence time of molten steel in the tundish by incorporation of flow modifiers like dams, weirs, turbo-pad, etc. These flow modifiers also help in reducing the dead or short circuit zones within the tundish which is significant for maintaining thermal and chemical homogeneity of molten steel. Thus, it becomes important to analyze the flow of molten steel in the tundish for different configuration of flow modifiers. In the present work, effect of varying positions and heights/depths of dam and weir on the dead volume in tundish is studied. Steady state thermal and flow profiles of molten steel within the tundish are obtained using OpenFOAM. Subsequently, Residence Time Distribution analysis is performed to obtain the percentage of dead volume in the tundish. Design of Experiment method is then used to configure different tundish geometries for varying positions and heights/depths of dam and weir, and dead volume for each tundish design is obtained. A second-degree polynomial with two-term interactions of independent variables to predict the dead volume in the tundish with positions and heights/depths of dam and weir as variables are computed using Multiple Linear Regression model. This polynomial is then used in an optimization framework to obtain the optimal tundish geometry for minimizing dead volume using Sequential Quadratic Programming optimization.

Keywords: design of experiments, multiple linear regression, OpenFOAM, residence time distribution, sequential quadratic programming optimization, steel, tundish

Procedia PDF Downloads 180

Authors: Imam Muthohar, Budi Hartono, Sigit Priyanto, Hardiansyah Hardiansyah

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The eruption of Merapi Volcano in Yogyakarta, Indonesia in 2010 caused many casualties due to minimum preparedness in facing disaster. Increasing population capacity and evacuating to safe places become very important to minimize casualties. Regional government through the Regional Disaster Management Agency has divided disaster-prone areas into three parts, namely ring 1 at a distance of 10 km, ring 2 at a distance of 15 km and ring 3 at a distance of 20 km from the center of Mount Merapi. The success of the evacuation is fully supported by road network infrastructure as a way to rescue in an emergency. This research attempts to model evacuation process based on the rise of refugees in ring 1, expanded to ring 2 and finally expanded to ring 3. The model was developed using SATURN (Simulation and Assignment of Traffic to Urban Road Networks) program version 11.3. 12W, involving 140 centroid, 449 buffer nodes, and 851 links across Yogyakarta Special Region, which was aimed at making a preliminary identification of road networks considered vulnerable to disaster. An assumption made to identify vulnerability was the improvement of road network performance in the form of flow and travel times on the coverage of ring 1, ring 2, ring 3, Sleman outside the ring, Yogyakarta City, Bantul, Kulon Progo, and Gunung Kidul. The research results indicated that the performance increase in the road networks existing in the area of ring 2, ring 3, and Sleman outside the ring. The road network in ring 1 started to increase when the evacuation was expanded to ring 2 and ring 3. Meanwhile, the performance of road networks in Yogyakarta City, Bantul, Kulon Progo, and Gunung Kidul during the evacuation period simultaneously decreased in when the evacuation areas were expanded. The results of preliminary identification of the vulnerability have determined that the road networks existing in ring 1, ring 2, ring 3 and Sleman outside the ring were considered vulnerable to the evacuation of Mount Merapi eruption. Therefore, it is necessary to pay a great deal of attention in order to face the disasters that potentially occur at anytime.

Keywords: model, evacuation, SATURN, vulnerability

Procedia PDF Downloads 143

Authors: Akinsola Oluwatosin, Udofia Samuel, Odofin Mayowa

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The study is aimed at assessing the Geographic Information System (GIS)-based spatial analysis of Survival Pattern and Treatment Outcomes of Multi-Drug Resistant Tuberculosis (MDR-TB) cases for Lagos, Nigeria, with an objective to inform priority areas for public health planning and resource allocation. Multi-drug resistant tuberculosis (MDR-TB) develops due to problems such as irregular drug supply, poor drug quality, inappropriate prescription, and poor adherence to treatment. The shapefile(s) for this study were already georeferenced to Minna datum. The patient’s information was acquired on MS Excel and later converted to . CSV file for easy processing to ArcMap from various hospitals. To superimpose the patient’s information the spatial data, the addresses was geocoded to generate the longitude and latitude of the patients. The database was used for the SQL query to the various pattern of the treatment. To show the pattern of disease spread, spatial autocorrelation analysis was used. The result was displayed in a graphical format showing the areas of dispersing, random and clustered of patients in the study area. Hot and cold spot analysis was analyzed to show high-density areas. The distance between these patients and the closest health facility was examined using the buffer analysis. The result shows that 22% of the points were successfully matched, while 15% were tied. However, the result table shows that a greater percentage of it was unmatched; this is evident in the fact that most of the streets within the State are unnamed, and then again, most of the patients are likely to supply the wrong addresses. MDR-TB patients of all age groups are concentrated within Lagos-Mainland, Shomolu, Mushin, Surulere, Oshodi-Isolo, and Ifelodun LGAs. MDR-TB patients between the age group of 30-47 years had the highest number and were identified to be about 184 in number. The outcome of patients on ART treatment revealed that a high number of patients (300) were not ART treatment while a paltry 45 patients were on ART treatment. The result shows the Z-score of the distribution is greater than 1 (>2.58), which means that the distribution is highly clustered at a significance level of 0.01.

Keywords: tuberculosis, patients, treatment, GIS, MDR-TB

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Authors: Tanjil Sowgat, Shamim Kobir

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World is now experiencing the consequences of climate change such as increased heat stress due to high temperature rise. In the context of changing climate, this study intends to find out the planning interventions necessary to adapt to the current heat stress in the planned residential areas of Khulna city. To carry out the study Sonadanga residential area (phase I) of Khulna city has been taken as the study site. This residential neighbourhood covering an area of 30 acres has 206 residential plots. The study area comprises twelve access roads, one park, one playfield, one water body and two street furniture’s. This study conducts visual analysis covering green, open space, water body, footpath, drainage and street trees and furniture and questionnaire survey deals with socio-economic, housing tenancy, experience of heat stress and urban design interventions. It finds that the current state that accelerates the heat stress condition such as lack of street trees and inadequate shading, maximum uses are not within ten minutes walking distance, no footpath for the pedestrians and lack of well-maintained street furniture. It proposes that to adapt to the heat stress pedestrian facilities, buffer sidewalk with landscaping, street trees and open spaces, soft scape, natural and man-made water bodies, green roofing could be effective urban design interventions. There are evidences of limited number of heat stress adaptive planned residential area. Since current sub-division planning practice focuses on rigid land use allocation, it partly addresses the climatic concerns through creating open space and street trees. To better respond to adapt to the heat stress, urban design considerations in the context of sub-division practice would bring more benefits.

Keywords: climate change, urban design, adaptation, heat stress, water-logging

Procedia PDF Downloads 272

Authors: S. Junaid S. Qazi, Denice C. Bay, Raymond Chew, Raymond J. Turner

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EmrE, a member of the small multidrug resistance protein family in bacteria is considered to be the archetypical member of its family. It confers host resistance to a wide variety of quaternary cation compounds (QCCs) driven by proton motive force. Generally, purification yield is a challenge in all membrane proteins because of the difficulties in their expression, isolation and solubilization. EmrE is extremely hydrophobic which make the purification yield challenging. We have purified EmrE protein using two different approaches: organic solvent membrane extraction and hexahistidine (his6) tagged Ni-affinity chromatographic methods. We have characterized changes present between ligand affinity of untagged and his6-tagged EmrE proteins in similar membrane mimetic environments using biophysical experimental techniques. Purified proteins were solubilized in a buffer containing n-dodecyl-β-D-maltopyranoside (DDM) and the conformations in the proteins were explored in the presence of four QCCs, methyl viologen (MV), ethidium bromide (EB), cetylpyridinium chloride (CTP) and tetraphenyl phosphonium (TPP). SDS-Tricine PAGE and dynamic light scattering (DLS) analysis revealed that the addition of QCCs did not induce higher multimeric forms of either proteins at all QCC:EmrE molar ratios examined under the solubilization conditions applied. QCC binding curves obtained from the Trp fluorescence quenching spectra, gave the values of dissociation constant (Kd) and maximum specific one-site binding (Bmax). Lower Bmax values to QCCs for his6-tagged EmrE shows that the binding sites remained unoccupied. This lower saturation suggests that the his6-tagged versions provide a conformation that prevents saturated binding. Our data demonstrate that tagging an integral membrane protein can significantly influence the protein.

Keywords: small multidrug resistance (SMR) protein, EmrE, integral membrane protein folding, quaternary ammonium compounds (QAC), quaternary cation compounds (QCC), nickel affinity chromatography, hexahistidine (His6) tag

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Authors: Michelle R. Sullivan

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Positive romantic relationships and marriages offer a buffer against a host of physical and emotional difficulties. Conversely, poor relationship quality and marital discord can have deleterious consequences for individuals and families. Research has described non-monogamy, infidelity, and consensual non-monogamy, as both consequential and causal of relationship difficulty, or as a unique way a couple strives to make a relationship work. Much research on consensual non-monogamy has built on feminist theory and critique. To the author’s best knowledge, to date, no studies have examined the predictive relationship between individual and relationship characteristics and expectations of non-monogamy. The current longitudinal study: 1) estimated the prevalence of expectations of partner non-monogamy and 2) evaluated whether gender, sexual identity, age, education, how a couple met, and relationship quality were predictive expectations of partner non-monogamy. This study utilized the publically available longitudinal dataset, How Couples Meet and Stay Together. Adults aged 18- to 98-years old (n=4002) were surveyed by phone over 5 waves from 2009-2014. Demographics and how a couple met were gathered through self-report in Wave 1, and relationship quality and expectations of partner non-monogamy were gathered through self-report in Waves 4 and 5 (n=1047). The prevalence of expectations of partner non-monogamy (encompassing both infidelity and consensual non-monogamy) was 4.8%. Logistic regression models indicated that sexual identity, gender, education, and relationship quality were significantly predictive of expectations of partner non-monogamy. Specifically, male gender, lower education, identifying as lesbian, gay, or bisexual, and a lower relationship quality scores were predictive of expectations of partner non-monogamy. Male gender was not predictive of expectations of partner non-monogamy in the follow up logistic regression model. Age and whether a couple met online were not associated with expectations of partner non-monogamy. Clinical implications include awareness of the increased likelihood of lesbian, gay, and bisexual individuals to have an expectation of non-monogamy and the sequelae of relationship dissatisfaction that may be related. Future research directions could differentiate between non-monogamy subtypes and the person and relationship variables that lead to the likelihood of consensual non-monogamy and infidelity as separate constructs, as well as explore the relationship between predicting partner behavior and actual partner behavioral outcomes.

Keywords: open relationship, polyamory, infidelity, relationship satisfaction

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Authors: Ali Ayatic, Emad Mozaffari, Bahareh Tanhaei, Maryam Khajenoori, Saeedeh Movaghar Khoshkho, Ali Ayati

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The abuse of antibiotics, such as tetracycline (TC), is a great global threat to people and the use of topical antibiotics is a promising tact that can help to solve this problem. Antibiotic therapy is often appropriate and necessary for acute wound infections, while topical tetracycline can be highly efficient in improving the wound healing process in diabetics. Due to the advantages of drug-loaded hydrogels as wound dressing, such as ease of handling, high moisture resistance, excellent biocompatibility, and the ability to activate immune cells to speed wound healing, it was found as an ideal wound treatment. In this work, the tetracycline-loaded hydrogels combining agar (AG) and κ-carrageenan (k-CAR) as polymer materials were prepared, in which span60 surfactant was introduced inside as a drug carrier. The Field Emission Scanning Electron Microscopes (FESEM) and Fourier-transform infrared spectroscopy (FTIR) techniques were employed to provide detailed information on the morphology, composition, and structure of fabricated drug-loaded hydrogels and their mechanical properties, and hydrogel permeability to water vapor was investigated as well. Two types of gram-negative and gram-positive bacteria were used to explore the antibacterial properties of prepared tetracycline-contained hydrogels. Their swelling and drug release behavior was studied using the changing factors such as the ratio of polysaccharides (MAG/MCAR), the span60 surfactant concentration, potassium chloride (KCl) concentration and different release media (deionized water (DW), phosphate-buffered saline (PBS), and simulated wound fluid (SWF)) at different times. Finally, the kinetic behavior of hydrogel swelling was studied. Also, the experimental data of TC release to DW, PBS, and SWF using various mathematical models such as Higuchi, Korsmeyer-Peppas, zero-order, and first-order in the linear and nonlinear modes were evaluated.

Keywords: drug release, hydrogel, tetracycline, wound healing

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Authors: Bruna Lima Gomes, Vanessa Caroline De Barros Bonato, Luciano Freschi, Eduardo Purgatto

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Plant hormones are long known to be tightly associated with fruit development and are involved in controlling various aspects of fruit ripening. For fleshy fruits, ripening is characterized for changes in texture, color, aroma and other parameters that markedly contribute to its quality. Ethylene is one of the major players regulating the ripening-related processes, but emerging evidences suggest that auxin is also part of this dynamic control. Thus, the aim of this study was providing new insights into the auxin role during ripening and the hormonal interplay between auxin and ethylene. For that, tomato fruits (Micro-Tom) were collected at mature green stage and separated in four groups: one for indole-3-acetic acid (IAA) treatment, one for ethylene, one for a combination of IAA and ethylene, and one for control. Hormone solution was injected through the stylar apex, while mock samples were injected with buffer only. For ethylene treatments, fruits were exposed to gaseous hormone. Then, fruits were left to ripen under standard conditions and to assess ripening development, hue angle was reported as color indicator and ethylene production was measured by gas chromatography. The transcript levels of three ripening-related ethylene receptors (LeETR3, LeETR4 and LeETR6) were evaluated by RT-qPCR. Results showed that ethylene treatment induced ripening, stimulated ethylene production, accelerated color changes and induced receptor expression, as expected. Nonetheless, auxin treatment showed the opposite effect once fruits remained green for longer time than control group and ethylene perception has changed, taking account the reduced levels of receptor transcripts. Further, treatment with both hormones revealed that auxin effect in delaying ripening was predominant, even with higher levels of ethylene. Altogether, the data suggest that auxin modulates several aspects of the tomato fruit ripening modifying the ethylene perception. The knowledge about hormonal control of fruit development will help design new strategies for effective manipulation of ripening regarding fruit quality and brings a new level of complexity on fruit ripening regulation.

Keywords: ethylene, auxin, fruit ripening, hormonal crosstalk

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Authors: Yun-Chin Chung, Nileema Divate, Gen-Hung Chen, Pei-Ru Huang, Rupesh Divate

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Many environmental factors, such as glucose concentration, ethanol, temperature, osmotic pressure and pH, decrease the production rate of ethanol using yeast as a starter. Fermentation starters with high tolerance to various stresses are always demanded for brewing industry. Trehalose, a storage carbohydrate in cell wall of yeast, plays an important role in tolerance of environmental stress by preserving integrity of plasma membrane and stabilizing proteins. Furan aldehydes are toxic to yeast and the growth rate of yeast is significantly reduced if furan aldehydes were present in the fermentation medium. In yeast, aldehyde reductase is involved in the detoxification of reactive aldehydes and consequently the growth of yeast is improved. The aims of this study were to construct a genetic recombinant Saccharomyces cerevisiae or Pichia pastoris with furfural and HMF degrading and high ethanol tolerance capacities. Yeast strains were engineered by genetic recombination for overexpression of trehalose-6-phosphate synthase gene (tps1) and aldehyde reductase gene (ari1). TPS1 gene was cloned from S. cerevisiae by reverse transcription-polymerase chain reaction (RT-PCR) and then ligated with pGAPZαC vector. The constructed vector, pGAPZC-tps1, was transformed to recombinant yeasts strain with overexpression of ari1. The transformants with pGAPZC-tps1-ari1 were generated called STA (S. cerevisiae) and PTA (P. pastoris) with overexpression of tps1, ari1. PCR with tps1-specific primers and western blot with his-tag confirmed the gene insertion and protein expression of tps1 in the transformants, respectively. The neutral trehalase gene (nth1) of STA was successfully deleted and the novel strain STAΔN will be used for further study, including the measurement of trehalose concentration and ethanol, furfural tolerance assay.

Keywords: genetic recombinant, yeast, ethanol tolerance, trehalase, aldehyde reductase

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Authors: Zahra Soleimani Rad, Fariborz Masoudi, Shirin Tondkar

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Turquoise is one of the most well-known gemstones in Iran. The mineralogy, crystallography, and gemology of Shahr-e-Babak turquoise in Kerman were investigated and the results are presented in this research. The Miduk porphyry copper deposit is positioned in the Shahr-Babak area in Kerman province, Iran. This deposit is located 85 km NW of the Sar-Cheshmeh porphyry copper deposit. Preliminary mineral exploration was carried out from 1967 to 1970. So far, more than fifty diamond drill holes, each reaching a maximum depth of 1013 meters, have provided evidence supporting the presence of significant and promising porphyry copper mineralization at the Miduk deposit. The mineral deposit harbors a quantity of 170 million metric tons of ore, characterized by a mean composition of 0.86% copper (Cu), 0.007% molybdenum (Mo), 82 parts-per-billion gold (Au), and 1.8 parts-per-million silver (Ag). The Supergene enrichment layer, which constitutes the predominant source of copper ore, exhibits an approximate thickness of 50 meters. Petrography shows that the texture is homogeneous. In terms of a gemstone, greasy luster and blue color are seen, and samples are similar to what is commonly known as turquoise. The geometric minerals were detected in XRD analysis by analyzing the data using the x-pert software. From the mineralogical point of view; the turquoise gemstones of Miduk of Kerman consist of turquoise, quartz, mica, and hübnerite. In this article, to our best knowledge, we are stating the hübnerite mineral identified and seen in the Persian turquoise. Based on the obtained spectra, the main mineral of the Miduk samples from the six members of the turquoise family is the turquoise type with identical peaks that can be used as a reference for identification of the Miduk turquoise. This mineral is structurally composed of phosphate units, units of Al, Cu, water, and hydroxyl units, and does not include a Fe unit. In terms of gemology, the quality of a gemstone depends on the quantity of the turquoise phase and the amount of Cu in it according to SEM and XRD analysis.

Keywords: turquoise, hübnerite, XRD analysis, Miduk, Kerman, Iran

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Authors: Abhishek N. Srivastava, Rahul Singh, Sumedha Chakma

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The rapid urbanization in the developing countries is generating an enormous amount of waste leading to the creation of unregulated landfill sites at various places at its disposal. The liquid waste, known as leachate, produced from these landfills sites is severely affecting the surrounding water quality. The water quality in the proximity areas of the landfill is found affected by various physico-chemical parameters of leachate such as pH, alkalinity, total hardness, conductivity, chloride, total dissolved solids (TDS), total suspended solids (TSS), sulphate, nitrate, phosphate, fluoride, sodium and potassium, biological parameters such as biochemical oxygen demand (BOD), chemical oxygen demand (COD), Faecal coliform, and heavy metals such as cadmium (Cd), lead (Pb), iron (Fe), mercury (Hg), arsenic (As), cobalt (Co), manganese (Mn), zinc (Zn), copper (Cu), chromium (Cr), nickel (Ni). However, all these parameters are distributive in leachate that produced according to the nature of waste being dumped at various landfill sites, therefore, it becomes very difficult to predict the main responsible parameter of leachate for water quality contamination. The present study is endeavour the comparative analysis of the physical, chemical and biological parameters of various landfills in India viz. Okhla landfill, Ghazipur landfill, Bhalswa ladfill in NCR Delhi, Deonar landfill in Mumbai, Dhapa landfill in Kolkata and Kodungayaiyur landfill, Perungudi landfill in Chennai. The statistical analysis of the parameters was carried out using the Statistical Packages for the Social Sciences (SPSS) and LandSim 2.5 model to simulate the long term effect of various parameters on different time scale. Further, the uncertainties characterization of various input parameters has also been analysed using fuzzy alpha cut (FAC) technique to check the sensitivity of various water quality parameters at the proximity of numerous landfill sites. Finally, the study would help to suggest the best method for the prevention of pollution migration from the landfill sites on priority basis.

Keywords: landfill leachate, water quality, LandSim, fuzzy alpha cut

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Authors: Constanza Monterrubio-Solis, Jose Tomas Ibarra

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The fertile interdependence of social-ecological systems reveals itself in the interactions between native forests and seeds, home gardens, kitchens, foraging activities, local knowledge, and food practices, creating particular flavors and food meanings as part of cultural identities within territories. Resilience in local-food systems, from a relational perspective, can be understood as the balance between persistence and adaptability to change. Food growing, preparation, and consumption are constantly changing and adapting as expressions of agency of female and male indigenous peoples and peasants. This paper explores local food systems’ expressions of resilience in the la Araucanía region of Chile, namely: diversity, redundancy, buffer capacity, modularity, self-organization, governance, learning, equity, and decision-making. Applying ethnographic research methods (participant observation, focus groups, and semi-structured interviews), this work reflects on the experience developed through work with Mapuche women cultivating home gardens in the region since 2012; it looks to material and symbolic elements of resilience in the local indigenous food systems. Local food systems show indeed indicators of social-ecological resilience. The biocultural memory is expressed in affection to particular flavors and recipes, the cultural importance of seeds and reciprocity networks, as well as an accurate knowledge about the indicators of the seasons and weather, which have allowed local food systems to thrive with a strong cultural foundation. Furthermore, these elements turn into biocultural resistance in the face of the current institutional pressures for rural specialization, processes of cultural assimilation such as agroecosystems and diet homogenization, as well as structural threats towards the diversity and freedom of native seeds. Thus, the resilience-resistance dynamic shown by the social-ecological systems of the southern Andes is daily expressed in the local food systems and flavors and is key for diverse and culturally sound social-ecological health.

Keywords: biocultural heritage, indigenous food systems, social-ecological resilience, southern Andes

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Authors: Grzegorz Swiderski, Agata Jablonska-Trypuc, Natalia Popow, Renata Swislocka, Wlodzimierz Lewandowski

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Doxorubicin belongs to the group of anthracycline antitumor antibiotics. Because of the wide spectrum of actions, it is one of the most widely used anthracycline antibiotics, including the treatment of breast, ovary, bladder, lung cancers as well as neuroblastoma, lymphoma, leukemia and myeloid leukemia. Antitumor activity of doxorubicin is based on the same mechanisms as for most anthracyclines. Like the metal ions affect the nucleic acids on many biological processes, so the environment of the metal chelates of antibiotics can have a significant effect on the pharmacological properties of drugs. Complexation of anthracyclines with metal ions may contribute to the production of less toxic compounds. In the framework of this study, the composition of complexes obtained in aqueous solutions of doxorubicin with metal ions (Cu2+ and Zn2+). Complexation was analyzed by spectrophotometric titration in aqueous solution at pH 7.0. The pH was adjusted with 0.02M Tris-HCl buffer. The composition of the complexes found was Cu: doxorubicin (1: 2) and a Zn: doxorubicin (1: 1). The effect of Dox, Dox-Cu and Dox-Zn was examined in MCF-7 breast cancer cell line, which were obtained from American Type Culture Collection (ATCC). The compounds were added to the cultured cells for a final concentration in the range of 0,01µM to 0,5µM. The number of MCF-7 cells with division into living and dead, was determined by direct counts of cells with the use of trypan blue dye using LUNA Logos Biosystems cell counter. ApoTox-Glo Triplex Assay (Promega, Madison, Wisconsin, USA) was used according to the manufacturer’s instructions to measure the MCF-7 cells’ viability, cytotoxicity and apoptosis. We observed a decrease in cells proliferation in a dose-dependent manner. An increase in cytotoxicity and decrease in viability in the ApoTox Triplex assay was also showed for all tested compounds. Apoptosis, showed as caspase 3/7 activation, was observed only in Dox treatment. In Dox-Zn and Dox-Cu caspase 3/7 activation was not observed. This work was financially supported by National Science Centre, Poland, under the research project number 2014/13/B/NZ7/02 352.

Keywords: anticancer properties, anthracycline antibiotic, doxorubicine, metal complexes

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Authors: Mark White

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The best practices for owners and urban planners to manage tsunami risk have evolved during the last fifty years, and related technical advances have created opportunities for them to obtain better performance than in earlier cataclysmic tsunami inundations. This basic pattern is illustrated at Hilo Bay, the waterfront area of Hilo, Hawaii, an urban seaport which faces the most severe tsunami hazard of the Hawaiian archipelago. Since April 1, 1946, Hilo Bay has endured tsunami waves with a maximum water height exceeding 2.5 meters following four severe earthquakes: Unimak Island (Mw 8.6, 6.1 m) in 1946; Valdiva (Mw 9.5, the largest earthquake of the 20th century, 10.6 m) in 1960; William Prince Sound (Mw 9.2, 3.8 m) in 1964; and Kalapana (Mw 7.7, the largest earthquake in Hawaii since 1868, 2.6 m) in 1975. Ignoring numerous smaller tsunamis during the same time frame, these four cataclysmic tsunamis have caused property losses in Hilo to exceed $1.25 billion and more than 150 deaths. It is reasonable to foresee another cataclysmic tsunami inundating the urban core of Hilo in the next 50 years, which, if unchecked, could cause additional deaths and losses in the hundreds of millions of dollars. Urban planners and individual owners are now in a position to reduce these losses in the next foreseeable tsunami that generates maximum water heights between 2.5 and 10 meters in Hilo Bay. Since 1946, Hilo planners and individual owners have already created buffer zones between the shoreline and its historic downtown area. As these stakeholders make inevitable improvements to the built environment along and adjacent to the shoreline, they should incorporate new methods for better managing the obvious tsunami risk at Hilo. At the planning level, new manmade land forms, such as tsunami parks and inundation reservoirs, should be developed. Individual owners should require their design professionals to include sacrificial seismic and tsunami fuses that will perform well in foreseeable severe events and that can be easily repaired in the immediate aftermath. These investments before the next cataclysmic tsunami at Hilo will yield substantial reductions in property losses and fatalities.

Keywords: hilo, tsunami parks, reservoirs, fuse systems, risk managment

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Authors: Soledad Carinelli, Iñigo Fernández, José Luis González-Mora, Pedro A. Salazar-Carballo

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Mastitis is the most relevant inflammatory disease in cattle, affecting the animal health and causing important economic losses on dairy farms. This disease takes place in the mammary gland or udder when some opportunistic microorganisms, such as Staphylococcus aureus, Streptococcus agalactiae, Corynebacterium bovis, etc., invade the teat canal. According to the severity of the inflammation, mastitis can be classified as sub-clinical, clinical and chronic. Standard methods for mastitis detection include counts of somatic cells, cell culture, electrical conductivity of the milk, and California test (evaluation of “gel-like” matrix consistency after cell lysed with detergents). However, these assays present some limitations for accurate detection of subclinical mastitis. Currently, haptoglobin, an acute phase protein, has been proposed as novel and effective biomarker for mastitis detection. In this work, an electrochemical biosensor based on polydopamine-modified magnetic nanoparticles (MNPs@pDA) for haptoglobin detection is reported. Thus, MNPs@pDA has been synthesized by our group and functionalized with hemoglobin due to its high affinity to haptoglobin protein. The protein was labeled with specific antibodies modified with alkaline phosphatase enzyme for its electrochemical detection using an electroactive substrate (1-naphthyl phosphate) by differential pulse voltammetry. After the optimization of assay parameters, the haptoglobin determination was evaluated in milk. The strategy presented in this work shows a wide range of detection, achieving a limit of detection of 43 ng/mL. The accuracy of the strategy was determined by recovery assays, being of 84 and 94.5% for two Hp levels around the cut off value. Milk real samples were tested and the prediction capacity of the electrochemical biosensor was compared with a Haptoglobin commercial ELISA kit. The performance of the assay has demonstrated this strategy is an excellent and real alternative as screen method for sub-clinical bovine mastitis detection.

Keywords: bovine mastitis, haptoglobin, electrochemistry, magnetic nanoparticles, polydopamine

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Authors: Rafay Ahmed, Condon Lau

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Bone healing is a complex and sequential process involving changes at the molecular level. Raman spectroscopy is a promising technique to study bone mineral and matrix environments simultaneously. In this study, subcritical calvarial defects are used to study bone composition during healing without discomposing the fracture. The model allowed to monitor the natural healing of bone avoiding mechanical harm to the callus. Calvarial defects were created using 1mm burr drill in the parietal bones of Sprague-Dawley rats (n=8) that served in vivo defects. After 7 days, their skulls were harvested after euthanizing. One additional defect per sample was created on the opposite parietal bone using same calvarial defect procedure to serve as control defect. Raman spectroscopy (785 nm) was established to investigate bone parameters of three different skull surfaces; in vivo defects, control defects and normal surface. Principal component analysis (PCA) was utilized for the data analysis and interpretation of Raman spectra and helped in the classification of groups. PCA was able to distinguish in vivo defects from normal surface and control defects. PC1 shows that the major variation at 958 cm⁻¹, which corresponds to ʋ1 phosphate mineral band. PC2 shows the major variation at 1448 cm⁻¹ which is the characteristic band of CH2 deformation and corresponds to collagens. Raman parameters, namely, mineral to matrix ratio and crystallinity was found significantly decreased in the in vivo defects compared to surface and controls. Scanning electron microscope and optical microscope images show the formation of newly generated matrix by means of bony bridges of collagens. Optical profiler shows that surface roughness increased by 30% from controls to in vivo defects after 7 days. These results agree with Raman assessment parameters and confirm the new collagen formation during healing.

Keywords: Raman spectroscopy, principal component analysis, calvarial defects, tissue characterization

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Authors: Rim Ben Amor, Myriam Abidi, Moncef Gueddari

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The purpose of the present study is to assess the level and the distribution of CaO, SO3, Cd, Cu, Pb and Zn incore sediments of Ghannouch-Gabes coast, Gulf of Gabes, Tunisian Mediterranean coast. The XRD analyses indicate that the sediments of Ghannouch-Gabes coast are mainly composed of quartz, calcite, gypsum and fluorine reflecting the impact of the phosphate fertilizer industrial waste. The vertical distribution of surface sediments shows for all the elements analyzed, that the area located between the commercial and the fishing port of Gabes, is the most polluted zone, where the two harbors acted as barriers and limited the dispersion of phosphogypsum discharge. The abundance order of metals was found to be Zn > Cd > Cu >Pb and that the highest levels of heavy metals were found in the uppermost segment of the sediment core compared to lower depth subsurface due to a continuous input of PG release and showed that the area between the two harbor suffered from several types of pollutants compared to reference core C1, collected from non-industrialized area. The level of pollution was evaluated using contamination factor (Cf), pollution load index (PLI) and the geoaccumulation index (Igeo). The obtained results of Igeo allowed us to distinguish that the area between the commercial harbor of Ghannouch and the fishing harbor of Gabes is the most polluted where sediments are strongly contaminated for Pb, Cu and Cd. The pollution load index (PLI) of all sediments collected classified them as "polluted". According to contamination factor (Cf), the sediments can be considered as ‘considerable’ to ‘very high’ contaminated for Pb, ‘very high to moderate’ for Cd, ‘ moderate’ for Zn, between ‘moderate’ and ‘considerable’ for Cu. Statistical analyses show that heavy metals, fluoride, calcium and sulphate are resulting from the same anthropogenic origin. The metallic pollution status of sediments of Ghanouch -Gabes coast is worrying and requires a serious intervention.

Keywords: trace metals, phosphogypsum, core sediments, accumulation factor, contamination factor

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Authors: Ebrahim Izadi-Darbandi, Masoud Azad, Masumeh Dehghan

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In order to study the effects of nitrogen and phosphoruse management and wheat sowing method on wheat yield, two experiments was performed as factorial, based on completely randomized design with three replications at Research Farm, Faculty of Agriculture, Ferdowsi University of Mashhad, Iran in 2009. In the first experiment nitrogen application rates (100kg ha-1, 200 kg ha-1, 300 kg ha-1), phosphorus application rates (100 kg ha-1, 200 kg ha-1) and two levels of their application methods (Broadcast and Band) were studied. The second experiment treatments included of wheat sowing methods (single-row with 30 cm distance and twine row on 60 cm width ridges), as main plots and nitrogen and phosphorus application methods (Broadcast and Band) as sub plots (150 kg ha-1). Phosphorus and nitrogen sources for fertilization at both experiment were respectively super phosphate, applied before wheat sowing and incorporated with soil and urea, applied in two phases (50% pre plant) and (50%) near wheat shooting. Results from first experiment showed that the effect of fertilizers application methods were significant (p≤0.01) on wheat yield increasing. Band application of phosphorus and nitrogen were increased biomass and seed yield of wheat with nine and 15% respectively compared to their broadcast application. The interaction between the effects of nitrogen and phosphorus application rate with phosphorus and nitrogen application methods, showed that band application of fertilizers and the rate of application of 200kg/ha phosphorus and 300kg/ha nitrogen were the best methods in wheat yield improvement. The second experiment also showed that the effect of wheat sowing method and fertilizers application methods were significant (p≤0.01) on wheat seed and biomass yield improvement. Wheat twine row on 60 cm width ridges sowing method, increased its biomass and seed yield for 22% and 30% respectively compared to single-row with 30 cm. Wheat sowing method and fertilizers application methods interaction indicated that band application of fertilizers and wheat twine row on 60 cm width ridges sowing method was the best treatment on winter wheat yield improvement. In conclusion these results indicated that nitrogen and phosphorus management in wheat and modifying wheat sowing method have important role in increasing fertilizers use efficiency.

Keywords: band application, broadcast application, rate of fertilizer application, wheat seed yield, wheat biomass yield

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Authors: Zhala Ahmad, Zainab Lazim, Haider Hamzah

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Bacterial resistance is a pressing global health issue, with multidrug-resistant (MDR), extensively drug-resistant (XDR), and pandrug-resistant (PDR) strains to pose a serious threat. In this context, researchers are investigating effective, safe, and affordable metabolites to combat these pathogens. This study focuses on gum essential oil (GEO) extracted from Pistacia atlantica and its activity and the mechanism of action against XDR Gram-negative Acinetobacter baumannii. GEO was extracted by hydrodistillation and analyzed using GC-MS. Eleven A. baumannii isolates were collected from the ward environment of Burn and Plastic Surgery Hospital in Al Sulaymaniyah City, Iraq. They were identified using the VITEK 2 system, 16S rRNA gene, and confirmed with the blaₒₓₐ₋₅₁ gene; A. baumannii ATCC 19606 was used as a reference strain. The isolates were identified as resistant to twelve different antibiotics spanning six distinct antibiotic classes while showing susceptibility to tetracycline and trimethoprim. Over 40 chemical constituents were detected in the gum's essential oils, with α-pinene being the most abundant. GEO was found to inhibit the growth of A. baumannii isolates; the minimum inhibitory concentration (MIC) of GEO was 2.5 µl/ml. GEO induced protein leakage, phosphate, and potassium ion efflux, distorted cell morphology, and cell death in the tested bacteria. GEO exhibited bacterial clearance and anti-adhesion activity using Band-Aids. This study's findings suggest that GEO could be used as a potential alternative treatment for infectious diseases caused by XRD pathogens, shedding further light on the importance of GEO in biomedical applications. Future studies must focus on generating clinically feasible sources of GEO for testing in small animal models before proceeding to human trials, ensuring safe and effective translation from the laboratory to the clinic.

Keywords: antibiotic resistance, Acinetobacter baumannii, essential oils, Pistacia atlantica, alpha-pinene

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Authors: N. B. Ikenweiwe, A. A. Alimi, N. A. Bamidele, O. A. Ewumi, K. Fasina, S. O. Otubusin

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A study on the physical, chemical and biological parameters of the lower course of Ogun River, Isheri-Olofin was carried out between January and December 2014 in order to determine the effects of the anthropogenic activities of the Kara abattoir and domestic waste depositions on the quality of the water. Water samples were taken twice each month at three selected stations A, B and C (based on characteristic features or activity levels) along the water course. Samples were analysed using standard methods for chemical and biological parameters the same day in the laboratory while physical parameters were determined in-situ with water parameters kit. Generally, results of Transparency, Dissolved Oxygen, Nitrates, TDS and Alkalinity fall below the permissible limits of WHO and FEPA standards for drinking and fish production. Results of phosphates, lead and cadmium were also low but still within the permissible limit. Only Temperature and pH were within limit. Low plankton community, (phytoplankton, zooplankton), which ranges from 3, 5 to 40, 23 were as a result of low levels of DO, transparency and phosphate. The presence of coliform bacteria of public health importance like Escherichia coli, Proteus vulgaris, Aeromonas sp., Shigella sp, Enterobacter aerogenes as well as gram negative bacteria Proteus morganii are mainly indicators of faecal pollution. Fish and other resources obtained from this water stand the risk of being contaminated with these organisms and man is at the receiving end. The results of the physical, chemical and some biological parameters of Isheri, Ogun River, according to this study showed that the live forms of aquatic and fisheries resources there are dwelling under stress as a result of deposition of bones, horns, faecal components, slurry of suspended solids, fat and blood into the water. Government should therefore establish good monitoring system against illegal waste depositions and create education programmes that will enlighten the community on the social, ecological and economic values of the river.

Keywords: damage, ecosystem, human activities, Isheri ogun river

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Authors: Mohamed Trigui, Fatma Masmoudi, Imen Zouari

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Massive utilizations of chemical fertilizer and chemical pesticides in agriculture sector to improve the farming productivity have created increasing environmental damages. Then, agriculture must become sustainable, focusing on production systems that respect the environment and help to reduce climate change. Isolation and microbial identification of new bacterial strains from naturally saline habitats and compost extracts could be a prominent way in pest management and crop production under saline conditions. In this study, potential mechanisms involved in plant growth promotion and suppressive activity against fungal diseases of a compost extract produced from poultry manure/olive husk compost and halotolerant and halophilic bacterial strains under saline stress were investigated. On the basis of the antimicrobial tests, different strains isolated from Sfax solar saltern (Tunisia) and from compost extracts were selected and tested for their plant growth promoting traits, such as siderophores production, nitrogen fixation, phosphate solubilization and the production of extracellular hydrolytic enzymes (protease and lipase) under in-vitro conditions. Among 450 isolated bacterial strains, 16 isolates showed potent antifungal activity against the tested plant pathogenic fungi. Their identification based on 16S rRNA gene sequence revealed they belonged to different species. Some of these strains were also characterized for their plant growth promoting capacities. Obtained results showed the ability of four strains belonging to Bacillus genesis to ameliorate germination rate and root elongation compared to the untreated positive controls. Combinatorial capacity of halotolerant bacteria with antimicrobial activity and plant growth promoting traits could be promising sources of interesting bioactive substances under saline stress.

Keywords: abiotic stress, biofertilizer, biotic stress, compost extract, halobacteria, plant growth promoting (PGP), soil fertility

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Authors: Yuda Purwana Roswanjaya, Wouter Kohlen, Rene Geurts

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The Trema genus represents a group of fast-growing tropical tree species within the Cannabaceae. Interestingly, five species nested in this lineage -known as Parasponia- can establish rhizobium nitrogen-fixing root nodules, similar to those found in legumes. Parasponia and legumes use a conserved genetic network to control root nodule formation, among which are genes also essential for mycorrhizal symbiosis (the so-called common symbiotic pathway). However, Trema species lost several genes that function exclusively in nodulation, suggesting a loss-of the nodulation trait in Trema. Strikingly, in a Trema orientalis population found in Malaysian Borneo we identified a truncated SYMBIOSIS RECEPTOR KINASE (SYMRK) mutant allele lacking a large portion of the c-terminal kinase domain. In legumes this gene is essential for nodulation and mycorrhization. This raises the question whether Trema orientalis can still be mycorrhized. To answer this question, we established quantitative mycorrhization assay for Parasponia andersonii and Trema orientalis. Plants were grown in closed pots on half strength Hoagland medium containing 20 µM potassium phosphate in sterilized sand and inoculated with 125 spores of Rhizopagus irregularis (Agronutrion-DAOM197198). Mycorrhization efficiency was determined by analyzing the frequency of mycorrhiza (%F), the intensity of the mycorrhizal colonization (%M) and the arbuscule abundance (%A) in the root system. Trema orientalis RG33 can be mycorrhized, though with lower efficiency compared to Parasponia andersonii. From this we conclude that a functional SYMRK kinase domain is not essential for Trema orientalis mycorrhization. In ongoing experiments, we aim to investigate the role of SYMRK in Parasponia andersonii mycorrhization and nodulation. For this two Parasponia andersonii symrk CRISPR-Cas9 mutant alleles were created. One mimicking the TorSYMRKRG33 allele by deletion of exon 13-15, and a full Parasponia andersonii SYMRK knockout.

Keywords: endomycorrhization, Parasponia andersonii, symbiosis receptor kinase (SYMRK), Trema orientalis

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Authors: Sungmin Park, Gyungmok Nam, Seungpyo Woo, Young Choi, Sangheon Park, Sang-Hee Yoon

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Puncturing human skin with microneedles is critically important for microneedle-mediate drug delivery. Despite of extensive efforts in the past decades, the scale-up fabrication of sharp-tipped and high-aspect-ratio microneedles, especially made of biodegradable polymers, is still a long way off. Here, we present a UV dose insensitive and cost-effective microfabrication method for the biodegradable polymer microneedles with sharp tips and long lengths which can pierce human skin with low insertion force. The biodegradable polymer microneedles are fabricated with the polymer solution casting where a poly(lactic-co-glycolic acid) (PLGA, 50:50) solution is coated onto a SU-8 mold prepared with a reverse, ramped, and rotational (R3) UV exposure. The R3 UV exposure is modified from the multidirectional UV exposure both to suppress UV reflection from the bottom surface without anti-reflection layers and to optimize solvent concentration in the SU-8 photoresist, therefore achieving robust (i.e., highly insensitive to UV dose) and cost-effective fabrication of biodegradable polymer microneedles. An optical model for describing the spatial distribution of UV irradiation dose of the R3 UV exposure is also developed to theoretically predict the microneedle geometry fabricated with the R3 UV exposure and also to demonstrate the insensitiveness of microneedle geometry to UV dose. In the experimental characterization, the microneedles fabricated with the R3 UV exposure are compared with those fabricated with a conventional method (i.e., multidirectional UV exposure). The R3 UV exposure-based microfabrication reduces the end-tip radius by a factor of 5.8 and the deviation from ideal aspect ratio by 74.8%, compared with conventional method-based microfabrication. The PLGA microneedles fabricated with the R3 UV exposure pierce full-thickness porcine skins successfully and are demonstrated to completely dissolve in PBS (phosphate-buffered saline). The findings of this study will lead to an explosive growth of the microneedle-mediated drug delivery market.

Keywords: R³ UV exposure, optical model, UV dose, reflection, solvent concentration, biodegradable polymer microneedle

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Authors: Iraj Rezaei, Kamal Al Din Niknami

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Tapeh Zagheh is an important prehistoric site located in the central plateau of Iran, which has settlement layers of the Neolithic and Chalcolithic periods. For this research, 38 soil samples were collected from different parts of the site, as well as two samples from its outside as witnesses. Then the samples were analyzed by XRF. The purpose of this research was to identify some places with special usage for human activities in Tapeh Zagheh by measuring the amount of some special elements in the soil. The result of XRF analysis shows a significant amount of P and K in samples No.3 (fourth floor) and No.4 (third floor), probably due to certain activities such as food preparation and consumption. Samples No.9 and No.10 can be considered suitable examples of the hearths of the prehistoric period in the central plateau of Iran. The color of these samples was completely darkened due to the presence of ash, charcoal, and burnt materials. According to the XRF results, the soil of these hearths has very high amounts of elements such as P, Ca, Mn, S, K, and significant amounts of Ti, Fe, and Na. In addition, the elemental composition of sample No. 14, which was taken from a home waster, also has very high amounts of P, Mn, Mg, Ti, and Fe and high amounts of K and Ca. Sample No. 11, which is related to soil containing large amounts of waster of the kiln, along with a very strong increase in Cl and Na, the amount of elements such as K, Mg, and S has also increased significantly. It seems that the reason for the increase of elements such as Ti and Fe in some Tapeh Zagheh floors (for example, samples number 1, 2, 3, 4, 5) was the use of materials such as ocher mud or fire ash in the composition of these floors. Sample No. 13, which was taken from an oven located in the FIX trench, has very high amounts of Mn, Ti, and Fe and high amounts of P and Ca. Sample No. 15, which is related to House No. VII (probably related to a pen or a place where animals were kept) has much more phosphate compared to the control samples, which is probably due to the addition of animal excrement and urine to the soil. Sample No. 29 was taken from the north of the industrial area of Zagheh village (place of pottery kilns). The very low amount of index elements in sample No. 29 shows that the industrial activities did not extend to the mentioned point, and therefore, the range of this point can be considered as the boundary between the residential part of the Zagheh village and its industrial part.

Keywords: prehistory, multi-elemental analysis, Tapeh Zagheh, XRF

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Authors: S. Fröhlich, M. Herold, M. Allmer

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Early stage quantitative analysis of host cell protein (HCP) variations is challenging yet necessary for comprehensive bioprocess development. High resolution mass spectrometry (HRMS) provides a high-end technology for accurate identification alongside with quantitative information. Hereby we describe a flexible HRMS assay platform to quantify HCPs relevant in microbial expression systems such as E. Coli in both up and downstream development by means of MVDA tools. Cell pellets were lysed and proteins extracted, purified samples not further treated before applying the SMART tryptic digest kit. Peptides separation was optimized using an RP-UHPLC separation platform. HRMS-MSMS analysis was conducted on an Orbitrap Velos Elite applying CID. Quantification was performed label-free taking into account ionization properties and physicochemical peptide similarities. Results were analyzed using SIEVE 2.0 (Thermo Fisher Scientific) and SIMCA (Umetrics AG). The developed HRMS platform was applied to an E. Coli expression set with varying productivity and the corresponding downstream process. Selected HCPs were successfully quantified within the fmol range. Analysing HCP networks based on pattern analysis facilitated low level quantification and enhanced validity. This approach is of high relevance for high-throughput screening experiments during upstream development, e.g. for titer determination, dynamic HCP network analysis or product characterization. Considering the downstream purification process, physicochemical clustering of identified HCPs is of relevance to adjust buffer conditions accordingly. However, the technology provides an innovative approach for label-free MS based quantification relying on statistical pattern analysis and comparison. Absolute quantification based on physicochemical properties and peptide similarity score provides a technological approach without the need of sophisticated sample preparation strategies and is therefore proven to be straightforward, sensitive and highly reproducible in terms of product characterization.

Keywords: process analytical technology, mass spectrometry, process characterization, MVDA, pattern recognition

Procedia PDF Downloads 231