Search results for: GFP-LC3 dot formation assay

Authors: Jongwoo Lee, Dae-Eun Kang, Sang-Young Park

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This study presents a precise relative navigational method for satellites flying in formation using laser-based intermittent measurement data. The measurement data for the relative navigation between two satellites consist of a relative distance measured by a laser instrument and relative attitude angles measured by attitude determination. The relative navigation solutions are estimated by both the Extended Kalman filter (EKF) and unscented Kalman filter (UKF). The solutions estimated by the EKF may become inaccurate or even diverge as measurement outage time gets longer because the EKF utilizes a linearization approach. However, this study shows that the UKF with the appropriate scaling parameters provides a stable and accurate relative navigation solutions despite the long measurement outage time and large initial error as compared to the relative navigation solutions of the EKF. Various navigation results have been analyzed by adjusting the scaling parameters of the UKF.

Keywords: satellite relative navigation, laser-based measurement, intermittent measurement, unscented Kalman filter

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Authors: Maria Lucia G. Lourenco, Viviane Y. Hibaru, Keylla H. N. P. Pereira, Fabiana F. Souza, Joao C. P. Ferreira, Simone B. Chiacchio, Luiz H. A. Machado

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Isosporiasis is an affliction caused by coccidial protozoa belonging to genera Isospora spp. or Cystoisospora spp., which may parasitize the small and large intestines of dogs, of which neonates and young animals present higher risk of infection. This study aims at reporting a case of isosporiasis in neonate Pitbull dogs, as well as the diagnosis and treatment. Seven Pitbull puppies were admitted to the São Paulo State University (UNESP) Veterinary Hospital, Botucatu, São Paulo, Brazil, with history of yellowish diarrhea without mucus or blood for the past two days. The animals were five days old. The history of the mother, a primiparous two-year-old, revealed that she was properly vaccinated, not de-wormed and did not present diarrhea. The clinical examination revealed that the neonates weighted between 308 and 360 grams, and presented normal reflexes, moderate dehydration, body temperatures between 36.8 and 37.2 ºC, blood sugar between 103 and 124 mg/dL and normal appetite. A full blood count and a parasitology assay were performed to aid in the diagnosis. The full blood count detected eosinophilia, without any other relevant alterations. The parasitology assay (Willis-Molly & Faust) revealed the presence of Cystoisospora spp. The treatment was instituted with heated fluid therapy with Ringer’s Lactate (4 mL/100 g, subcutaneous) and antibiotic therapy with sulfamethoxazole-trimethoprim (15 mg/kg, orally) every 12 hours for ten days. The mother and other dogs that came in contact with the newborns were also treated. The environment was disinfected for 10 minutes with 1.6% quaternary ammonium. After 10 days, the newborns presented normal clinical signs and no alterations in the full blood count. Isosporiasis is an affliction with high mortality rates in litters that should be diagnosed and treated as soon as possible to increase the survival rates in these patients.

Keywords: Cystoisospora spp., neonatal infection, puppies, diarrhea,

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Authors: Annapurna Basavaraju, Arianna Mastrodonato, Franz Heitmeir

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The Advisory Council for Aeronautics Research in Europe (ACARE) is creating awareness for the overall reduction of NOx emissions by 80% in its vision 2020. Hence this promotes the researchers to work on novel technologies, one such technology is the use of alternative fuels. Among these fuels hydrogen is of interest due to its one and only significant pollutant NOx. The influence of NOx formation due to hydrogen combustion depends on various parameters such as air pressure, inlet air temperature, air to fuel jet momentum ratio etc. Appropriately, this research is motivated to investigate the impact of the air to fuel jet momentum ratio onto the NOx formation in a hydrogen combustion chamber for aircraft engines. The air to jet fuel momentum is defined as the ratio of impulse/momentum of air with respect to the momentum of fuel. The experiments were performed in an existing combustion chamber that has been previously tested for methane. Premix of the reactants has not been considered due to the high reactivity of the hydrogen and high risk of a flashback. In order to create a less rich zone of reaction at the burner and to decrease the emissions, a forced internal recirculation flow has been achieved by integrating a plate similar to honeycomb structure, suitable to the geometry of the liner. The liner has been provided with an external cooling system to avoid the increase of local temperatures and in turn the reaction rate of the NOx formation. The injected air has been preheated to aim at so called flameless combustion. The air to fuel jet momentum ratio has been inspected by changing the area of fuel inlets and keeping the number of fuel inlets constant in order to alter the fuel jet momentum, thus maintaining the homogeneity of the flow. Within this analysis, promising results for a flameless combustion have been achieved. For a constant number of fuel inlets, it was seen that the reduction of the fuel inlet diameter resulted in decrease of air to fuel jet momentum ratio in turn lowering the NOx emissions.

Keywords: combustion chamber, hydrogen, jet momentum, NOx emission

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Authors: Aisha Musaazi Sebunya Nakitto, Anika E. Wagner, Yusuf B. Byaruhanga, John H. Muyonga

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The prevalence and burden of diabetes are rapidly increasing globally, stemming from changes in lifestyle and dietary habits. Although several drugs are available to treat type 2 diabetes mellitus (T2DM), many are accompanied by several side effects and are often costly. Solanum anguivi Lam. fruits (SALF) are bitter berries that commonly grow in the wild and are traditionally cultivated by many globally as a remedy for T2DM. This effect is likely attributable to the presence of bioactive compounds such as phenolics, flavonoids, saponins, alkaloids, and vitamin C in SALF. In this study, we investigated the morphological characteristics of different SALF accessions, and the effect of ripeness stages and thermal treatments on the bioactive compounds contents (BCC) and antioxidant activity (AA) of SALF accessions. Using the fruit fly, Drosophila melanogaster (D. melanogaster) model, we explored the potential impact of dietary SALF in preventing and treating T2DM phenotypes. Morphological characterization was conducted based on descriptors of Solanum species. The BCC and AA of SALF at different ripeness stages (unripe, yellow, orange, and red) and after thermal treatments were determined using spectrophotometry, HPLC, and gravimetry. Male and female fruit flies were fed a high-sugar diet (HSD) to induce a T2DM-like phenotype, while control flies were fed on SY10 medium upto 24 days. Experimental flies were exposed to HSD supplemented with 5 or 10 mg/ml SALF. The therapeutic and prevention effect of SALF in T2DM-like phenotype were investigated on weight, climbing activity, glucose and triglyceride contents, survival, and gene expression of PPARγ co-activator 1α fly homolog Srl and Drosophila insulin-like peptides. Methods in fly studies included Gustatory assay, Climbing assay, Glucose GOD-PAP assay, Triglyceride GPO-PAP assay, Roti-Quant®, and Real Time-PCR analysis. Ripeness stage significantly influenced SALF BCC and AA, and this was dependent on the accession. The unripe stage had the highest AA and total phenolics and flavonoids; orange stage was rich in saponins, while the red stage had the highest alkaloid contents. Boiling and steaming increased the total phenolics and AA upto 4-fold and 3-fold, respectively. Drying at low temperature resulted in higher phenolics and AA than the control. In the therapeutic model, the HSD-fed female flies exhibited elevated glucose levels, which exhibited a dose-dependent reduction upon exposure to SALF-supplemented diet. Female flies fed on a SALF+ HSD exhibited a significant increase in survival compared to HSD-fed and control diet-fed flies. SALF supplementation did not alter weights, fitness, and triglyceride levels of female flies in comparison with HSD-only-fed flies. The mRNA levels of Srl decreased in HSD-fed flies compared to the control-fed, with no effect observed in females exposed to HSD+SALF. Similarly, in the preventative model, SALF diet resulted in higher survival of supplemented flies compared to controls. Consumption of boiled unripe SALF may result in the highest health benefits due to high phenolic contents and antioxidant activity observed. Dietary intake of SALF significantly lowered glucose levels and increased survival of the D. melanogaster model. Additional studies in higher organisms are needed to explore the preventative and therapeutic potential of SALF in T2DM.

Keywords: solanum anguivi, type 2 diabetes mellitus, bioactive compounds, drosophila melanogaster, survival, antioxidant activity

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Authors: Marco Jan Rafael C. Sicam, Maria Daniella C. Yambao

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The Philippines is a tectonically active archipelagic country situated near the Circum-Pacific Belt. Hence, seismic hazard assessments are important in the nation-building. In 2014, the Philippines Institute of Volcanology and Seismology (PHIVOLCS) mapped a 12-km NW-trending unnamed active fault near Boso-Boso River, Rizal. Given the limited nature of their technical report, they would like to further consolidate relevant data about this fault. As such, this study aims to characterize the surface and subsurface expression of the fault along Boso-Boso River using rangefront morphology, structural criteria, and ground penetrating radar. This fault is subdivided into two segments: the first segment located in the city of Antipolo is mainly manifested in the upper Kinabuan Formation and terminating near Mt. Qutago, and the second segment in Baras, Pinugay, Rizal cuts through recent fluvial deposits and to the Guadalupe Formation. IfSAR-derived DTM data reveals the morphological expression of the fault defined by offset streams and ridges, linear sidehill valleys, and linear valleys. Fault gouges, fault breccia, transtentional flower structures, slickensides, and other shear sense markers observed in the units of the upper Cretaceous Kinabuan Formation indicate a sinistral sense of displacement. GPR radargram profiles revealed the presence of displacement in reflectors at 3-5 meters below the surface which may be suggestive of the fault within the area. Finally, the fault in Boso-Boso river may be a segment of the larger sinistral Montalban Fault in the north or largely affected by the movement from the Marikina Valley Fault System.

Keywords: NW unnamed fault, range-front morphology, shear sense markers, ground penetrating radar, boso-boso river, antipolo

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Authors: Rima Zakzouk, Yasushi Shimada, Yuan Zhou, Yasunori Sumi, Junji Tagami

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Background and Purpose: Swept source optical coherence tomography (OCT) is an interferometric imaging technique that has been recently used in cariology. In spite of progress made in adhesive dentistry, the composite restoration has been failing due to secondary caries which occur due to environmental factors in oral cavities. Therefore, a precise assessment to effective marginal sealing of restoration is highly required. The aim of this study was evaluating gap formation at composite/cavity walls interface with or without phosphoric acid etching using SS-OCT. Materials and Methods: Round tapered cavities (2×2 mm) were prepared in three locations, mid-coronal, cervical, and root of bovine incisors teeth in two groups (SE and PA Groups). While self-etching adhesive (Clearfil SE Bond) was applied for the both groups, Group PA had been already pretreated with phosphoric acid etching (K-Etchant gel). Subsequently, both groups were restored by Estelite Flow Quick Flowable Composite Resin. Following 5000 thermal cycles, three cross-sectionals were obtained from each cavity using OCT at 1310-nm wavelength at 0°, 60°, 120° degrees. Scanning was repeated after two months to monitor the gap progress. Then the average percentage of gap length was calculated using image analysis software, and the difference of mean between both groups was statistically analyzed by t-test. Subsequently, the results were confirmed by sectioning and observing representative specimens under Confocal Laser Scanning Microscope (CLSM). Results: The results showed that pretreatment with phosphoric acid etching, Group PA, led to significantly bigger gaps in mid-coronal and cervical compared to SE group, while in the root cavity no significant difference was observed between both groups. On the other hand, the gaps formed in root’s cavities were significantly bigger than those in mid-coronal and cervical within the same group. This study investigated the effect of phosphoric acid on gap length progress on the composite restorations. In conclusions, phosphoric acid etching treatment did not reduce the gap formation even in different regions of the tooth. Significance: The cervical region of tooth was more exposing to gap formation than mid-coronal region, especially when we added pre-etching treatment.

Keywords: image analysis, optical coherence tomography, phosphoric acid etching, self-etch adhesives

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Authors: Liaqat Ali, Hubert E. Blum, Türkân Sakinc

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Enterococcus faecium is an emerging multidrug-resistant nosocomial pathogen increased dramatically worldwide and causing bacteremia, endocarditis, urinary tract and surgical site infections in immunocomprised patients. The capsular polysaccharides that contribute to pathogenesis through evasion of the host innate immune system are also involved in hindering leukocyte killing of enterococci. The gene cluster (enterococcal polysaccharide antigen) of E. faecalis encoding homologues of many genes involved in polysaccharide biosynthesis. We identified two putative loci with 22 kb and 19 kb which contained 11 genes encoding for glycosyltransferases (GTFs); this was confirmed by using genome comparison of already sequenced strains that has no homology to known capsule genes and the epa-locus. The polysaccharide-conjugate vaccines have rapidly emerged as a suitable strategy to combat different pathogenic bacteria, therefore, we investigated a polysaccharide biosynthesis CapD protein in E. faecium contains 336 amino acids and had putative function for N-linked glycosylation. The deletion/knock-out capD mutant was constructed and complemented by homologues recombination method and confirmed by using PCR and sequencing. For further characterization and functional analysis, in-vitro cell culture and in-vivo a mouse infection models were used. Our ΔcapD mutant shows a strong hydrophobicity and all strains exhibited biofilm production. Subsequently, the opsonic activity was tested in an opsonophagocytic assay which shows increased in mutant compared complemented and wild type strains but more than two fold decreased in colonization and adherence was seen on surface of uroepithelial cells. However, a significant higher bacterial colonialization was observed in capD mutant during animal bacteremia infection. Unlike other polysaccharides biosynthesis proteins, CapD does not seems to be a major virulence factor in enterococci but further experiments and attention is needed to clarify its function, exact mechanism and involvement in pathogenesis of enteroccocal nosocomial infections eventually to develop a vaccine/ or targeted therapy.

Keywords: E. faecium, pathogenesis, polysaccharides, biofilm formation

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Authors: M. Sepahvand, M. Khorushy

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Gladiolus, being a cormous plant, it is principally propagated by the natural multiplication of new corms and cormels. In order to obtain callus from segmented corm which was obtained from in vitro culture, callus formation media were MS media supplemented with 4 levels of hormones such as 1.0 mg l-1 NAA + 0.5 mg l-1 BAP, 0.5 mg l-1 NAA + 0.25 mg l-1 BAP, 1.0 mg l-1 2, 4-D + 0.5 mg l-1 BAP, and 0.5 mg l-1 2, 4-D + 0.25 mg l-1 BAP. The results showed that the most weight of callus (2.28 g) was produced in MS callus formation media which were supplemented with 1.0 mg l-1 NAA + 0.5 mg l-1 BAP. This experiment was carried out in randomized completely design with 3 replications and each treatment with six jars. In second experiment for regeneration of callus, a factorial experiment in the form of randomized complete design with 12 treatments and 3 replications and each replication with six jars was carried out. The treatments consisted of callus culture media in 4 levels and regeneration culture media in 3 levels [control (no PGRs), MS with 0.2 mg l-1 BAP + 0.1 mg l-1 Kin + 0.01 mg l-1 NAA, and MS with 0.2 mg l-1 BAP + 0.05 mg l-1 Kin + 0.01 mg l-1 NAA]. The results showed that the best regeneration media were MS media which were supplemented with 0.2 mg l-1 BAP + 0.1 mg l-1 Kin. + 0.01 mg l-1 NAA that had the highest number of shoots (7/83 N), and shoot length (7/3 cm).

Keywords: regeneration, Segmented corm explant, callus, in vitro, gladiolus cv. white prosperity

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Authors: Venoos Iman, Suzita Mohd Noor, Syam Mohan, Mohamad Ibrahim Noordin

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Girinimbine, a carbazole alkaloid was isolated from the stem bark and root of Murraya koenigii and its structure and purity was identified by HPLC and LC-MS. Here we report that Girinimbine strongly inhibit angiogenesis activity both in vitro and in vivo. MTT result showed that girinimbine inhibits cell proliferation of the HUVECS cell line in vitro. Result of endothelial cell invasion, migration, tube formation and wound healing assays also demonstrated significant time and does dependent inhibition by girinimbine. Moreover, girinibine mediates its anti-angiogenic activity through up- and down-regulation of angiogenic and anti-aniogenic proteins. Furthermore, anti-angiogenic potential of girinimbine was evidenced in vivo on zebrafish model. Girinimbine inhibited neo-vessels formation in zebrafish embryos during 24 hours exposure time. Together, these results demonstrated for the first time that girinimbine could effectively suppress angiogenesis and strongly suggest that it might be a novel angiogenesis inhibitor.

Keywords: anti-angiogenic, carbazole alkaloid, girinimbine, zebrafish

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Authors: Liang-Jen Wang, Sung-Chou Li, Yuan-Ming Yeh, Sheng-Yu Lee, Ho-Chang Kuo, Chia-Yu Yang

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Background: Dysbiosis in the gut microbial community might be involved in the pathophysiology of attention deficit/hyperactivity disorder (ADHD). The fungal component of the gut microbiome, namely the mycobiota, is a hyperdiverse group of multicellular eukaryotes that can influence host intestinal permeability. This study therefore aimed to investigate the impact of fungal mycobiome dysbiosis and intestinal permeability on ADHD. Methods: Faecal samples were collected from 35 children with ADHD and from 35 healthy controls. Total DNA was extracted from the faecal samples, and the internal transcribed spacer (ITS) regions were sequenced using high-throughput next-generation sequencing (NGS). The fungal taxonomic classification was analysed using bioinformatics tools, and the differentially expressed fungal species between the ADHD and healthy control groups were identified. An in vitro permeability assay (Caco-2 cell layer) was used to evaluate the biological effects of fungal dysbiosis on intestinal epithelial barrier function. Results: The β-diversity (the species diversity between two communities), but not α-diversity (the species diversity within a community), reflected the differences in fungal community composition between ADHD and control groups. At the phylum level, the ADHD group displayed a significantly higher abundance of Ascomycota and significantly lower abundance of Basidiomycota than the healthy control group. At the genus level, the abundance of Candida (especially Candida albicans) was significantly increased in ADHD patients compared to the healthy controls. In addition, the in vitro cell assay revealed that C. albicans secretions significantly enhanced the permeability of Caco-2 cells. Conclusions: The current study is the first to explore altered gut mycobiome dysbiosis using the NGS platform in ADHD. The findings from this study indicated that dysbiosis of the fungal mycobiome and intestinal permeability might be associated with susceptibility to ADHD.

Keywords: ADHD, fungus, gut–brain axis, biomarker, child psychiatry

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Authors: Christian Fercher, Jiaul Islam, Simon R. Corrie

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Fluorescence-based immunodiagnostics are an emerging field in biosensor development and exhibit several advantages over traditional detection methods. While various affinity biosensors have been developed to generate a fluorescence signal upon sensing varying concentrations of analytes, reagentless, reversible, and continuous monitoring of complex biological samples remains challenging. Here, we aimed to genetically engineer biosensors based on single-chain antibody fragments (scFv) that are site-specifically labeled with environmentally sensitive fluorescent unnatural amino acids (UAA). A rational design approach resulted in quantifiable analyte-dependent changes in peak fluorescence emission wavelength and enabled antigen detection in vitro. Incorporation of a polarity indicator within the topological neighborhood of the antigen-binding interface generated a titratable wavelength blueshift with nanomolar detection limits. In order to ensure continuous analyte monitoring, scFv candidates with fast binding and dissociation kinetics were selected from a genetic library employing a high-throughput phage display and affinity screening approach. Initial rankings were further refined towards rapid dissociation kinetics using bio-layer interferometry (BLI) and surface plasmon resonance (SPR). The most promising candidates were expressed, purified to homogeneity, and tested for their potential to detect biomarkers in a continuous microfluidic-based assay. Variations of dissociation kinetics within an order of magnitude were achieved without compromising the specificity of the antibody fragments. This approach is generally applicable to numerous antibody/antigen combinations and currently awaits integration in a wide range of assay platforms for one-step protein quantification.

Keywords: antibody engineering, biosensor, phage display, unnatural amino acids

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Authors: Ahmed H. Abdullah, Pavel Kroupa

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Globular clusters (GC) are important objects for tracing the early evolution of a galaxy. We study the correlation between the cluster population and the global properties of the host galaxy. We found that the correlation between cluster population (NGC) and the baryonic mass (Mb) of the host galaxy are best described as 10 −5.6038Mb. In order to understand the origin of the U -shape relation between the GC specific frequency (SN) and Mb (caused by the high value of SN for dwarfs galaxies and giant ellipticals and a minimum SN for intermediate mass galaxies≈ 1010M), we derive a theoretical model for the specific frequency (SNth). The theoretical model for SNth is based on the slope of the power-law embedded cluster mass function (β) and different time scale (Δt) of the forming galaxy. Our results show a good agreement between the observation and the model at a certain β and Δt. The model seems able to reproduce higher value of SNth of β = 1.5 at the midst formation time scale.

Keywords: galaxies: dwarf, globular cluster: specific frequency, number of globular clusters, formation time scale

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Authors: Gulnara Galeeva

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The Analytic Hierarchy Process is widely used in the economic and financial studies, including the formation of investment portfolios. In this study, a generalized method of obtaining a vector of priorities for the case with separate pairwise comparisons of the expert opinion being presented as a set of several equal evaluations on a ratio scale is examined. The author claims that this method allows solving an important and up-to-date problem of excluding vagueness and ambiguity of the expert opinion in the decision making theory. The study describes the authentic wide pairwise comparison matrix. Its application in the formation of the efficient investment portfolio of intangible assets of a small business enterprise with limited funding is considered. The proposed method has been successfully approbated on the practical example of a functioning dental clinic. The result of the study confirms that the wide pairwise comparison matrix can be used as a simple and reliable method for forming the enterprise investment policy. Moreover, a comparison between the method based on the wide pairwise comparison matrix and the classical analytic hierarchy process was conducted. The results of the comparative analysis confirm the correctness of the method based on the wide matrix. The application of a wide pairwise comparison matrix also allows to widely use the statistical methods of experimental data processing for obtaining the vector of priorities. A new method is available for simple users. Its application gives about the same accuracy result as that of the classical hierarchy process. Financial directors of small and medium business enterprises get an opportunity to solve the problem of companies’ investments without resorting to services of analytical agencies specializing in such studies.

Keywords: analytic hierarchy process, decision processes, investment portfolio, intangible assets

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Authors: Nuo Xu, Kok Hun Goh, Jeyatharan Kumarasamy

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Pile load testing is significant during foundation construction due to its traditional role of design validation and routine quality control of the piling works. In order to verify whether piles can take loadings at specified settlements, piles will have to undergo working load test where the test load should normally up to 150% of the working load of a pile. Selection or sampling of piles for the working load test is done subject to the number specified in Singapore National Annex to Eurocode 7 SS EN 1997-1:2010. This paper presents an innovative way to rationalize the number of pile load test by adopting statistical analysis approach and looking at the coefficient of variance of pile elastic modulus using a case study at Singapore Tuas depot. Results are very promising and have shown that it is possible to reduce the number of working load test without influencing the reliability and confidence on the pile quality. Moving forward, it is suggested that more load test data from other geological formations to be examined to compare with the findings from this paper.

Keywords: elastic modulus of pile under soil interaction, jurong formation, kentledge test, pile load test

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Authors: Raudone Lina, Raudonis Raimondas, Gaivelyte Kristina, Pukalskas Audrius, Janulis Valdimaras, Viskelis Pranas

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Sorbus L. species are widely distributed in the Northern hemisphere and have been used for medicinal purposes in various traditional medicine systems and as food ingredients. Various Sorbus L. raw materials, fruits, leaves, inflorescences, barks, possess diuretic, anti-inflammatory, hypoglycemic, anti-diarrheal and vasoprotective activities. Phenolics, to whom main pharmacological activities are attributed, are compounds of interest due to their notable antioxidant activity. The aim of this study was to determine the antioxidant profiles of fruits and leaves of selected Sorbus L. species (S. anglica, S. aria f. latifolia, S. arranensis, S. aucuparia, S. austriaca, S. caucasica, S. commixta, S. discolor, S. gracilis, S. hostii, S. semi-incisa, S. tianschanica) and to identify the phenolic compounds with potent contribution to antioxidant activity. Twenty two constituents were identified in Sorbus L. species using ultra high performance liquid chromatography coupled to quadruple and time-of-flight mass spectrometers (UPLC–QTOF–MS). Reducing activity of individual constituents was determined using high performance liquid chromatography (HPLC) coupled to post-column FRAP assay. Signicantly greatest trolox equivalent values corresponding up to 45% of contribution to antioxidant activity were assessed for neochlorogenic and chlorogenic acids, which were determined as markers of antioxidant activity in samples of leaves and fruits. Characteristic patterns of antioxidant profiles obtained using HPLC post-column FRAP assay significantly depend on specific Sorbus L. species and raw materials and are suitable for equivalency research of Sorbus L. fruits and leaves. Selecting species and target plant organs with richest phenolic composition and strongly expressed antioxidant power is the first step in further research of standardized extracts.

Keywords: FRAP, antioxidant, phenolic, Sorbus L., chlorogenic acid, neochlorogenic acid

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Authors: Csilla Dudás, Éva Böszörményi, Bence Kutus, István Pálinkó, Pál Sipos

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In this study the behavior of alpha-D-isosaccharinate (2-hydroxymethyl-3-deoxy-D-erythro-pentonate, ISA−) in alkaline medium in the presence of calcium was studied. At first the Ca–ISA system was studied by Ca-ion selective electrode (Ca-ISE) in neutral medium at T = 25 °C and I = 1 M NaCl to determine the formation constant of the CaISA+ monocomplex, which was found to be logK = 1.01 ± 0.01 for the reaction of Ca2+ + ISA– = CaISA+. In alkaline medium pH potentiometric titrations were carried out to determine the composition and stability constant of the complex(es) formed. It was found that in these systems above pH = 12.5 the predominant species is the CaISAOH complex. Its formation constant was found to be logK = 3.04 ± 0.05 for the reaction of Ca2+ + ISA– + H2O = CaISAOH + H+ at T = 25 °C and I = 1 M NaCl. Solubility measurements resulted in data consistent with those of the potentiometric titrations. Temperature dependent NMR spectra showed that the slow exchange range between the complex and the free ligand is below 5 °C. It was also showed that ISA– acts as a multidentate ligand forming macrochelate Ca-complexes. The structure of the complexes was determined by using ab initio quantum chemical calculations.

Keywords: Ca-ISE potentiometry, calcium complexes, isosaccharinate ion, NMR spectroscopy, pH potentiometry

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Authors: M. R. Vijayakumar, Sanjay Kumar Singh, Lakshmi, Hithesh Dewangan, Sanjay Singh

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Trans resveratrol (RES) is a natural molecule proved for cancer preventive and therapeutic activities devoid of any potential side effects. However, the therapeutic application of RES in disease management is limited because of its rapid elimination from blood circulation thereby low biological half life in mammals. Therefore, the main objective of this study is to enhance the circulation as well as therapeutic efficacy using PEGylated liposomes. D-α-tocopheryl polyethylene glycol 1000 succinate (vitamin E TPGS) is applied as steric surface decorating agent to prepare RES liposomes by thin film hydration method. The prepared nanoparticles were evaluated by various state of the art techniques such as dynamic light scattering (DLS) technique for particle size and zeta potential, TEM for shape, differential scanning calorimetry (DSC) for interaction analysis and XRD for crystalline changes of drug. Encapsulation efficiency and invitro drug release were determined by dialysis bag method. Cancer cell viability studies were performed by MTT assay, respectively. Pharmacokinetic studies were performed in sprague dawley rats. The prepared liposomes were found to be spherical in shape. Particle size and zeta potential of prepared formulations varied from 64.5±3.16 to 262.3±7.45 nm and -2.1 to 1.76 mV, respectively. DSC study revealed absence of potential interaction. XRD study revealed presence of amorphous form in liposomes. Entrapment efficiency was found to be 87.45±2.14 % and the drug release was found to be controlled up to 24 hours. Minimized MEC in MTT assay and tremendous enhancement in circulation time of RES PEGylated liposomes than its pristine form revealed that the stearic stabilized PEGylated liposomes can be an alternative tool to commercialize this molecule for chemopreventive and therapeutic applications in cancer.

Keywords: trans resveratrol, cancer nanotechnology, long circulating liposomes, bioavailability enhancement, liposomes for cancer therapy, PEGylated liposomes

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Authors: D. P. Houhoula, J. Papaparaskevas, S. Konteles, A. Dargenta, A. Farka, C. Spyrou, M. Ziaka, S. Koussisis, E. Charvalos

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Objectives: Nanotechnology is providing revolutionary opportunities for the rapid and simple diagnosis of many infectious diseases. Staphylococcus aureus, Listeria monocytogenes and Salmonella enteritis are important human pathogens. Diagnostic assays for bacterial culture and identification are time consuming and laborious. There is an urgent need to develop rapid, sensitive, and inexpensive diagnostic tests. In this study, a gold nanoprobe strategy developed and relies on the colorimetric differentiation of specific DNA sequences based approach on differential aggregation profiles in the presence or absence of specific target hybridization. Method: Gold nanoparticles (AuNPs) were purchased from Nanopartz. They were conjugated with thiolated oligonucleotides specific for the femA gene for the identification of members of Staphylococcus aureus, the mecA gene for the differentiation of Staphylococcus aureus and MRSA Staphylococcus aureus, hly gene encoding the pore-forming cytolysin listeriolysin for the identification of Listeria monocytogenes and the invA sequence for the identification of Salmonella enteritis. DNA isolation from Staphylococcus aureus Listeria monocytogenes and Salmonella enteritis cultures was performed using the commercial kit Nucleospin Tissue (Macherey Nagel). Specifically 20μl of DNA was diluted in 10mMPBS (pH5). After the denaturation of 10min, 20μl of AuNPs was added followed by the annealing step at 58oC. The presence of a complementary target prevents aggregation with the addition of acid and the solution remains pink, whereas in the opposite event it turns to purple. The color could be detected visually and it was confirmed with an absorption spectrum. Results: Specifically, 0.123 μg/μl DNA of St. aureus, L.monocytogenes and Salmonella enteritis was serially diluted from 1:10 to 1:100. Blanks containing PBS buffer instead of DNA were used. The application of the proposed method on isolated bacteria produced positive results with all the species of St. aureus and L. monocytogenes and Salmonella enteritis using the femA, mecA, hly and invA genes respectively. The minimum detection limit of the assay was defined at 0.2 ng/μL of DNA. Below of 0.2 ng/μL of bacterial DNA the solution turned purple after addition of HCl, defining the minimum detection limit of the assay. None of the blank samples was positive. The specificity was 100%. The application of the proposed method produced exactly the same results every time (n = 4) the evaluation was repeated (100% repeatability) using the femA, hly and invA genes. Using the gene mecA for the differentiation of Staphylococcus aureus and MRSA Staphylococcus aureus the method had a repeatability 50%. Conclusion: The proposed method could be used as a highly specific and sensitive screening tool for the detection and differentiation of Staphylococcus aureus Listeria monocytogenes and Salmonella enteritis. The use AuNPs for the colorimetric detection of DNA targets represents an inexpensive and easy-to-perform alternative to common molecular assays. The technology described here, may develop into a platform that could accommodate detection of many bacterial species.

Keywords: gold nanoparticles, pathogens, nanotechnology, bacteria

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Authors: Lixin Pei, Zhilong Huang, Wenzhe Gang

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Organic geochemistry of source rocks and oils in the Nanpu Sag, Bohai Bay basin was studied on the basis of the results of Rock-Eval and biomarker. The possible source rocks consist of the third member (Es₃) and the first member (Es₁) of Shahejie formation and the third member of Dongying Formation (Ed₃) in the Nanpu Sag. The Es₃, Es₁, and Ed₃ source rock intervals in the Nanpu Sag all have high organic-matter richness and are at hydrocarbon generating stage, which are regarded as effective source rocks. The three possible source rock intervals have different biomarker associations and can be differentiated by gammacerane/αβ C₃₀ hopane, ETR ([C₂₈+C₂₉]/ [C₂₈+C₂₉+Ts]), C₂₇ diasterane/sterane and C₂₇/C₂₉ steranes, which suggests they deposited in different environments. Based on the oil-source rock correlation, the shallow oils mainly originated from the Es₃ and Es₁ source rocks in the Nanpu Sag. Through hydrocarbon generation and expulsion history of the source rocks, trap development history and accumulation history, the shallow oils mainly originated from paleo-reservoirs in the Es₃ and Es₁ during the period of Neotectonism, and the residual paleo-reservoirs in the Es₃ and Es₁ would be the focus targets in the Nanpu Sag; Bohai Bay Basin.

Keywords: source rock, biomarker association, Nanpu Sag, Bohai Bay Basin

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Authors: Duangkamol Thitivesa

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This paper focuses on the use of project work as a pretext for applying the conventions of writing, or the correctness of mechanics, usage, and sentence formation, in a content-based class in a Rajabhat University. Its aim was to explore to what extent the student teachers’ academic achievement of the basic writing features against the 70% attainment target after the use of project is. The organization of work around an agreed theme in which the students reproduce language provided by texts and instructors is expected to enhance students’ correct writing conventions. The sample of the study comprised of 38 fourth-year English major students. The data was collected by means of achievement test and student writing works. The scores in the summative achievement test were analyzed by mean score, standard deviation, and percentage. It was found that the student teachers do more achieve of practicing mechanics and usage, and less in sentence formation. The students benefited from the exposure to texts during conducting the project; however, their automaticity of how and when to form phrases and clauses into simple/complex sentences had room for improvement.

Keywords: project-based learning, project work, writing conventions, academic achievement

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Authors: Arda Bagus M, Yehezkiel Festian P, Budianto Santoso

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The Tourism Area of Southern Malang is administratively located in the Regency of Malang, East Java Province, Indonesia and geographically is in a position between 112o17' - 112o57' E dan 7o44' - 8o26' S. Southern Malang consists of several sub-districts that directly borders with the Indian Ocean, such as Donomulyo, Bantur, Gedangan, Sumbermanjing, Tirto Yudo, and Ampel Gading. This area has a high geotourism potential because of the existence of geodiversity such as beaches, waterfalls, caves, and karst area. However, to the best of the authors’ knowledge, there is still no systematic data that informs the geotourism potentials to the public. The aim of this research is to complete the lack of data and then arrange it systematically so it can be used for both tourism and research purposes. Research methods such as field observation, literature study, and depth interview to local people have been implemented. Aspects reviewed by visiting the field are accommodation, transportation, and the feasibility of a place to be geotourism object. The primary data was taken in Sumbermanjing, Gedangan, Bantur, and Donomulyo sub-district. A literature study is needed to determine the regional geology of Southern Malang and as a comparison to new data obtained in the field. The results of the literature study show that southern Malang consists of three formations: Wonosari Formation, Mandalaka Formation, and River-swamps Sediment Formation with the age range of Oligocene to Quaternary. Depth interviews have been conducted by involving local people with the aim of knowing cultural-history in the research area. From this research, the geotourism object distribution map has been made. The map also includes Geotrek and basic geological information of each object. The results of this research can support the development of geotourism in Southern Malang.

Keywords: geodiversity, geology, geotourism, geotrek, southern Malang

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Authors: E. A. Asabina, V. I. Pet'kov, P. A. Mayorov, A. V. Markin, N. N. Smirnova, A. M. Kovalskii, A. A. Usenko

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The purpose of this work was to synthesize and investigate phase formation, structure and thermophysical properties of the phosphates M_0.5+xM'_xZr_2–x(PO₄)₃ (M – Cd, Sr, Pb; M' – Mg, Co, Mn). The compounds were synthesized by sol-gel method. The results showed formation of limited solid solutions of NZP/NASICON type. The crystal structures of triple phosphates of the compositions MMg_0.5Zr_1.5(PO₄)₃ were refined by the Rietveld method using XRD data. Heat capacity (8–660 K) of the phosphates Pb_0.5+xMg_xZr_2-x(PO₄)₃ (x = 0, 0.5) was measured, and reversible polymorphic transitions were found at temperatures, close to the room temperature. The results of Rietveld structure refinement showed the polymorphism caused by disordering of lead cations in the cavities of NZP/NASICON structure. Thermal expansion (298−1073 K) of the phosphates MMg_0.5Zr_1.5(PO₄)₃ was studied by XRD method, and the compounds were found to belong to middle and low-expanding materials. Thermal diffusivity (298–573 K) of the ceramic samples of phosphates slightly decreased with temperature increasing. As was demonstrated, the studied phosphates are characterized by the better thermophysical characteristics than widespread fire-resistant materials, such as zirconia and etc.

Keywords: NASICON, NZP, phosphate, structure, synthesis, thermophysical properties

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Authors: Woei-Shyan Lee, Shuo-Ling Chang

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The nanoindentation behaviour and phase transformation of annealed single-crystal silicon wafers are examined. The silicon specimens are annealed at temperatures of 250, 350 and 450ºC, respectively, for 15 minutes and are then indented to maximum loads of 30, 50 and 70 mN. The phase changes induced in the indented specimens are observed using transmission electron microscopy (TEM) and micro-Raman scattering spectroscopy (RSS). For all annealing temperatures, an elbow feature is observed in the unloading curve following indentation to a maximum load of 30 mN. Under higher loads of 50 mN and 70 mN, respectively, the elbow feature is replaced by a pop-out event. The elbow feature reveals a complete amorphous phase transformation within the indented zone, whereas the pop-out event indicates the formation of Si XII and Si III phases. The experimental results show that the formation of these crystalline silicon phases increases with an increasing annealing temperature and indentation load. The hardness and Young’s modulus both decrease as the annealing temperature and indentation load are increased.

Keywords: nanoindentation, silicon, phase transformation, amorphous, annealing

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Authors: S. Reshma Reghu, Shiburaj Sugathan, T. G. Nandu, K. B. Ramesh Kumar, Mathew Dan

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Bacterial diseases are considered to be one of the most prevalent health hazards in the developing world and many bacteria are becoming resistant to existing antibiotics making the treatment ineffective. Thus, it is necessary to find novel targets and develop new antibacterial drugs with a novel mechanism of action. The process of bacterial cell division is a novel and attractive target for new antibacterial drug discovery. FtsZ, a homolog of eukaryotic tubulin, is the major protein of the bacterial cell division machinery and is considered as an important antibacterial drug target. Zingiberaceae, the Ginger family consists of aromatic herbs with creeping rhizomes. Many of these plants have antimicrobial properties.This study aimed to determine the anti-bacterial activity of selected Zingiberaceous plants by targeting bacterial cell division protein, FtsZ. Essential oils and methanol extracts of Amomum ghaticum, Alpinia galanga, Kaempferia galanga, K. rotunda, and Zingiber officinale were tested to find its antibacterial efficiency using disc diffusion method against authentic bacterial strains obtained from MTCC (India). Essential oil isolated from A.galanga and Z.officinale were further assayed for FtsZ inhibition assay following non-radioactive malachite green-phosphomolybdate assay using E. coli FtsZ protein obtained from Cytoskelton Inc., USA. Z.officinale essential oil possess FtsZ inhibitory property. A molecular docking study was conducted with the known bioactive compounds of Z. officinale as ligands with the E. coli FtsZ protein homology model. Some of the major constituents of this plant like catechin, epicatechin, and gingerol possess agreeable docking scores. The results of this study revealed that several chemical constituents in Ginger plants can be utilised as potential source of antibacterial activity and it can warrant further investigation through drug discovery studies.

Keywords: antibacterial, FtsZ, zingiberaceae, docking

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Authors: Samar Saadeldin Abdelmotalab Omer, Ikram Mohammed Eltayeb Elsiddig, Amna Beshir Medani Ahmed, Saad Mohammed Hussein Ayoub

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Many herbal medicinal products are considered as potential hypocholesterolemic agents with encouraging safety profiles, however, only a limited amount of clinical research exists to support their efficacy. The present study was designed to compare the antihypercholesterolaemic and antioxidant activities of the crude ethanolic extracts of Citrus reticulata peel, Zingiber officinale rhizome, and Sesamum indicum seeds. Forty-five rats were used throughout the experiment, which were divided into nine groups, five rats in each as follows; normal control group (normal rats fed with standard normal diet), rats fed hypercholesterolemic diet consisting of 1% cholesterol and 10% saturated animal fat, which were further divided into eight groups; hypercholesterolemic control group (rats only fed hypercholesterolemic diet), groups 3,4,5,6,7, and 8 were given Citrus reticulata, Zingiber officinale, and Sesamum indicum ethanolic extracts at doses of (250mg/kg and 500mg/kg, respectively) orally; and group 9 rats were given atorvastatin (0.18mg/kg) orally as a reference antihypercholesterolaemic drug. Blood samples were obtained four weeks following treatment from the retro-orbital venous plexus after fasting overnight from all groups and the lipid profile (serum total cholesterol (TC), high-density-lipoprotein-cholesterol (HDL-C), low-density lipoprotein-cholesterol (LDL-C), and triglycerides levels) was measured and the risk ratio (TC/HDL-C) was assessed. The antioxidant activity of the three plant extracts was determined using DPPH free-radical assay. Results of in vivo and in vitro antihypercholesterolaemic and antioxidant assay, respectively, revealed that the three extracts possess comparable antioxidant and anti-hypercholesterolaemic activities.

Keywords: anti hypercholesterolemic effects, antioxidant activity, HDL, LDL, TC, TGs, citrus reticulata, sesamum indicum, zingiber officinale

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Authors: Shama Ranjan Barua, Tofazzal M. Rakib, Mohammad Alamgir Hossain, Tania Ferdushy, Sharmin Chowdhury

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Rotavirus is one of the main etiologies of neonatal diarrhea in bovine calves that causes significant economic loss in Bangladesh. The present study was carried out to investigate the pathology of neonatal enteritis in calves due to bovine rotavirus infection in south-eastern part of Bangladesh. Rotavirus was identified by using ELISA, RT-PCR (Reverse Transcription Polymerase Chain Reaction), real-time RT-PCR. We examined 12 dead calves with history of diarrhea during necropsy. Among 12 dead calves, in gross examination, 6 were found with pathological changes in intestine, 5 calves had congestion of small intestine and rest one had no distinct pathological changes. Intestinal contents and/or faecal samples of all dead calves were collected and examined to confirm the presence of bovine rotavirus A using Enzyme linked immunosorbant assay (ELISA), RT-PCR and real-time RT-PCR. Out 12 samples, 5 (42%) samples revealed presence of bovine rotavirus A in three diagnostic tests. The histopathological changes were found almost exclusively limited in the small intestine. The lesions of rotaviral enteritis ranged from slight to moderate shortening (atrophy) of villi in the jejunum and ileum with necrotic crypts. The villi were blunt and covered by immature epithelial cells. Infected cells, stained with Haematoxylin and Eosin staining method, showed characteristic syncytia and eosinophilc intracytoplasmic inclusion body. The presence of intracytoplasmic inclusion bodies in enterocytes is the indication of viral etiology. The presence of rotavirus in the affected tissues and/or lesions was confirmed by three different immunological and molecular tests. The findings of histopathological changes will be helpful in future diagnosis of rotaviral infection in dead calves.

Keywords: calves, diarrhea, pathology, rotavirus

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Authors: S. Sulochana Singha

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Dhiyan or Dehan is the name of the community and language spoken by the Koch-Rajbangshi people of Barak Valley of Assam. Ethnically, they are Mongoloids, and their language belongs to the Indo-Aryan language family. However, Dhiyan is absent in any classification of Indo-Aryan languages. So the classification of Dhiyan language under the Indo-Aryan language family is completely based on the shared typological features of the other Indo-Aryan languages. Typologically, Dhiyan is an agglutinating language, and it shares many features of Indo-Aryan languages like presence of aspirated voiced stops, non-tonal, verb-person agreement, adjectives as different word class, prominent tense and subject object verb word order. Reduplication is a productive word-formation process in Dhiyan. Besides it also expresses plurality, intensification, and distributive. Generally, reduplication in Dhiyan can be at the morphological or lexical level. Morphological reduplication in Dhiyan involves expressives which includes onomatopoeias, sound symbolism, idiophones, and imitatives. Lexical reduplication in the language can be formed by echo formations and word reduplication. Echo formation in Dhiyan is formed by partial repetition from the base word which can be either consonant alternation or vowel alternation. The consonant alternation is basically found in onset position while the alternation of vowel is basically found in open syllable particularly in final syllable. Word reduplication involves reduplication of nouns, interrogatives, adjectives, and numerals which further can be class changing or class maintaining reduplication. The process of reduplication can be partial or complete whether it is lexical or morphological. The present paper is an attempt to describe some aspects of the formation, function, and usage of reduplications in Dhiyan which is mainly spoken in ten villages in the Eastern part of Barak River in the Cachar District of Assam.

Keywords: Barak-Valley, Dhiyan, Indo-Aryan, reduplication

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Authors: Noryani Natasha Yahaya, Anas Ibrahim, Juraidah Ahmad, Azura Ahmad, Mohd Ikmal Fazlan Rosli, Zailan Ramli, Muhd Sidek Muhd Norhasri

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The subgrade formation is an important element of the railway structure which holds overall track stability. Conventional track maintenance involves many substructure component replacements, as well as track re-ballasting on a regular basis is partially contributed to the embankment's long-term settlement problem. For subgrade long-term stability analysis, the geophysical method is commonly being used to diagnose those hidden sources/mechanisms of track deterioration problems that the normal visual method is unable to detect. Electrical resistivity tomography (ERT) is one of the applicable geophysical tools that are helpful in railway subgrade inspection/track monitoring due to its flexibility and reliability of the analysis. The ERT was conducted at KM 23.0 of Pinang Tunggal track to investigate the subgrade of railway track through the characterization/mapping on track formation profiling which was directly generated using 2D analysis of Res2dinv software. The profiles will allow examination of the presence and spatial extent of a significant subgrade layer and screening of any poor contact of soil boundary. Based on the finding, there is a mix/interpretation/intermixing of an interlayer between the sub-ballast and the sand. Although the embankment track considered here is at no immediate risk of settlement effect or any failure, the regular monitoring of track’s location will allow early correction maintenance if necessary. The developed data of track formation clearly shows the similarity of the side view with the assessed track. The data visualization in the 2D section of the track embankment agreed well with the initial assumption based on the main element structure general side view.

Keywords: ground track, assessment, resistivity, geophysical railway, method

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Authors: Tatiana S. Ogneva

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Aluminum and nickel-based intermetallic compounds have attracted the attention of scientific community as promising materials for heat-resistant and wear-resistant coatings in such manufacturing areas as microelectronics, aircraft and rocket building and chemical industries. Magnetron sputtering makes possible to coat materials without formation of liquid phase and improves the mechanical and functional properties of nickel aluminides due to the possibility of nanoscale structure formation. The purpose of the study is the investigation of structure and properties of intermetallic coatings produced by magnetron sputtering technique. The feature of this work is the using of composite targets for sputtering, which were consisted of two semicircular sectors of cp-Ni and cp-Al. Plates of alumina, silicon, titanium and steel alloys were used as substrates. To estimate sputtering conditions on structure of intermetallic coatings, a series of samples were produced and studied in detail using scanning and transition electron microcopy and X-Ray diffraction. Besides, nanohardness and scratching tests were carried out. The varying parameters were the distance from the substrate to the target, the duration and the power of the sputtering. The thickness of the obtained intermetallic coatings varied from 0.05 to 0.5 mm depending on the sputtering conditions. The X-ray diffraction data indicated that the formation of intermetallic compounds occurred after sputtering without additional heat treatment. Sputtering at a distance not closer than 120 mm led to the formation of NiAl phase. Increase in the power of magnetron from 300 to 900 W promoted the increase of heterogeneity of the phase composition and the appearance of intermetallic phases NiAl, Ni₂Al₃, NiAl₃, and Al under the aluminum side, and NiAl, Ni₃Al, and Ni under the nickel side of the target. A similar trend is observed with increasing the distance of sputtering from 100 to 60 mm. The change in the phase composition correlates with the changing of the atomic composition of the coatings. Scanning electron microscopy revealed that the coatings have a nanoscale grain structure. In this case, the substrate material and the distance from the substrate to the magnetron have a significant effect on the structure formation process. The size of nanograins differs from 10 to 83 nm and depends not only on the sputtering modes but also on material of a substrate. Nanostructure of the material influences the level of mechanical properties. The highest level of nanohardness of the coatings deposited during 30 minutes on metallic substrates at a distance of 100 mm reached 12 GPa. It was shown that nanohardness depends on the grain size of the intermetallic compound. Scratching tests of the coatings showed a high level of adhesion of the coating to substrate without any delamination and cracking. The results of the study showed that magnetron sputtering of composite targets consisting of nickel and aluminum semicircles makes it possible to form intermetallic coatings with good mechanical properties directly in the process of sputtering without additional heat treatment.

Keywords: intermetallic coatings, magnetron sputtering, mechanical properties, structure

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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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