Search results for: tagged MRI

Authors: Mohammed Alenezy

Abstract:

The function of cardiac motion can be used as an indicator of the heart abnormality by evaluating longitudinal, circumferential, and Radial Strain of the left ventricle. In this paper, the Radial Strain and SR is studied using tagged MRI (tMRI) data during the cardiac cycle on the mid-ventricle level of the left ventricle. Materials and methods: The short-axis view of the left ventricle of five healthy human (three males and two females) and four healthy male rats were imaged using tagged magnetic resonance imaging (tMRI) technique covering the whole cardiac cycle on the mid-ventricle level. Images were processed using Image J software to calculate the left ventricle wall Radial Strain and radial SR. The left ventricle Radial Strain and radial SR were calculated at the mid-ventricular level during the cardiac cycle. The peak Radial Strain for the human and rat heart was 40.7±1.44, and 46.8±0.68 respectively, and it occurs at 40% of the cardiac cycle for both human and rat heart. The peak diastolic and systolic radial SR for human heart was -1.78 s-1 ± 0.02 s-1 and 1.10±0.08 s-1 respectively, while for rat heart it was -5.16± 0.23s-1 and 4.25±0.02 s-1 respectively. Conclusion: This results show the ability of the tMRI data to characterize the cardiac motion during the cardiac cycle including diastolic and systolic phases which can be used as an indicator of the cardiac dysfunction by estimating the left ventricle Radial Strain and radial SR at different locations of the cardiac tissue. This study approves the validity of the tagged MRI data to describe accurately the cardiac radial motion.

Keywords: left ventricle, radial strain, tagged MRI, cardiac cycle

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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: Han-Bin Kim, Sooim Shin, Moonsung Choi

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There is a growing interest in introducing a desired functional group on enzymes in the field of protein engineering. In here, various redox centers were newly created using histidine tag, which is widely used for protein purification, plus cobalt in one of cupredoxins, amicyanin. The coordination of Cobalt-His tag and reactivity of the Co²⁺ loaded His-tag also were characterized. 3xHis-tag, 6xHis-tag, and 9xHis-tag were introduced on amicyanin by site-directed mutagenesis, and then Co²⁺ was loaded on each His-tagged amicyanin. The spectral changes at 330 nm corresponding to cobalt binding on His-tag site indicated the binding ratio of 3xHis-tag, 6xHis-tag, and 9xHis-tag to cobalt as 1:1, 1:2, 1:3 respectively. Based on kinetic studies of binding cobalt to 3xHis-tag, 6xHis-tag, and 9xHis-tagged amicyanin, the nature of the sites was elucidated. In addition, internal electron transfer properties between Cu¹⁺ site and engineered site of amicyanin were determined. These results provide insight into improvement of metal coordination and alternation of the redox properties of metal as a new catalytic site on proteins.

Keywords: amicyanin, cobalt, histidine, protein engineering

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Authors: Zainul A. Khan, Malik Z. Abdin, Jawaid A. Khan

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Begomoviruses belonging to the family Geminiviridae, have single-stranded circular DNA genomes that are monopartite or bipartite. The large intergenic region (LIR) of the monopartite and common region (CR) of bipartite begomoviruses possess promoter activity in their genomes. In this study, we have characterized novel bidirectional promoters from Cotton leaf curl Burewala virus (CLCuBuV) genome using high-throughput software and analyzed with PlantCARE, PLACE, Cister and PlantPAN databases. The promoters (Rep and CP promoters) were assayed both in stable and transient expression systems in tobacco as well as cotton plants. Rep and CP-based promoters from the LIR sequence of CLCuBuV and 35S promoter of Cauliflower mosaic virus (CaMV) were tagged with β-glucuronidase (GUS) and green fluorescent protein (GFP) reporter genes to check the efficacy of the promoters. Histochemical staining of GUS in transformed tobacco (Nicotiana tabacum cv. Xanthi) leaves showed higher GUS expression driven by CLCuBuV Rep (complimentary sense) promoter as compared to conventional CaMV 35S promoter and CLCuBuV CP (virion sense) promoter, respectively. GUS activity in individual plant cells driven by CLCuBuV Rep, CLCuBuV CP, and CaMV 35S promoter were quantified through fluorometric GUS assay and reverse transcription quantitative real-time PCR (RT-qPCR). The expression level of GUS tagged with CLCuBuV Rep promoter in the transformed tobacco plants was obtained 2 to 4 fold higher than CaMV 35S promoter. When CLCuBuV CP promoter was used, lower expression level was monitored than that by CaMV 35S promoter. The expression of GFP-tagged with CLCuBuV promoters was also investigated through agroinfiltration. The CLCuBuV Rep promoters showed stronger consistent transient expression in the leaves of N. benthamiana, N. tabacum and Gossypium hirsutum plants when compared with CaMV 35S and CLCuBuV CP promoter.

Keywords: Begmovirus, bidirectional promoter, CaMV 35S promoter, GFP, GUS, qPCR

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Authors: Jae Bok Heo, Yun Mi Lee, Hee Rang Yun

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Several GTPases are required for ribosome biogenesis and assembly. We recently characterized rice (Oryza sativa) nuclear/nucleolar GTPase 2 (OsNug2), belonging to the YlqF/YawG family of GTPases, as playing a role in pre-60S ribosomal subunit maturation. To investigate the potential factors involved in regulating the function of OsNug2, yeast two-hybrid screens were carried out using OsNug2 as bait. Rice serine/threonine kinase 1 (OsSTK1) was identified as a potential interacting protein candidate. In vitro pull down and bimolecular fluorescence complementation assays confirmed the interaction between OsNug2 and OsSTK1, and like green fluorescent protein-tagged OsNug2, green fluorescent protein-tagged OsSTK1 was targeted to the nucleus of Arabidopsis protoplasts. OsSTK1 was not found to affect the GTP-binding activity of OsNug2; however, when recombinant OsSTK1 was included in OsNug2 assay reaction mixtures, OsSTK1 increased the GTPase activity of OsNug2. To test whether OsSTK1 phosphorylates OsNug2 in vitro, a kinase assay was performed. OsSTK1 was found to have weak autophosphorylation activity and strongly phosphorylated serine 209 of OsNug2. Yeast complementation testing resulted in a GAL::OsNug2(S209N) mutant-harboring yeast strain exhibiting a growth-defective phenotype on galactose medium at 39°C, divergent from that of a yeast strain harboring GAL::OsNug2. The intrinsic GTPase activity of mutant OsNug2(S209N) was found to be similar to that of OsNug2, was not fully enhanced upon weak binding of OsSTK1. Our findings reported here indicate that OsSTK1 functions as a positive regulator protein of OsNug2 by enhancing the GTPase activity of OsNug2, and that the phosphorylation of serine 209 of OsNug2 is essential for the complete function of OsNug2 in ribosome biogenesis.

Keywords: OsSTK1, OsNug2, GTPase activity, GTP binding activity, phosphorylation

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Authors: Gyorgy Deak, Marius C. Raischi, Lucian P. Georgescu, Tiberius M. Danalache, Elena Holban, Madalina G. Boboc, Monica Matei, Catalina Iticescu, Marius V. Olteanu, Stefan Zamfir, Gabriel Cornateanu

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At present, on the Lower Danube, different types of pressures have been identified that affect the anadromous sturgeons stocks with an impact that leads to their decline. This paper presents techniques and procedures used by Romanian experts in the tagging and monitoring of anadromous sturgeons, as well as unique results at international level obtained on the basis of an informational volume collected in over 7 years of monitoring on these species behavior (both for adults as well as for ultrasonically tagged juveniles) on the Lower Danube. The local impact of hydrotechnical constructions (bottom sill, maritime navigation channel), the global impact of the poaching phenomenon and the impact of the restocking programs with sturgeon juveniles were assessed. Thus, the bottom sill impact on the Bala branch, the Bastroe Channel (cross-border impact) and the poaching phenomenon at the level of the Lower Danube was analyzed on the basis of a unique informational volume obtained through the use of patented monitoring systems by the Romanian experts (DKTB respectively, DKMR-01T). At the same time, the results from the monitoring of ultrasonically tagged sturgeon juveniles from the 2015 repopulation program are presented. Conclusions resulting from research can ensure favorable premises for finding some conservation solutions for CITES-protected sturgeon species that have survived for millions of years, currently being 1 species on the brink of extinction - Russian sturgeon, 2 species in danger of extinction - Beluga sturgeon and Stellate sturgeon and 2 species already extinct from the Lower Danube, namely common sturgeon and ship sturgeon.

Keywords: Lower Danube, sturgeons monitoring (adults and juveniles), tagging, impact on conservation

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Authors: Hwang Ahreum, Kang Taejoon, Kim Bongsoo

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Nanosensors for high sensitive detection of diseases have been widely studied to improve the quality of life. Here, we suggest robust nano-plasmonic diagnostic sensor using cysteine tagged protein G (Cys3-protein G) and ultraflat, ultraclean and single-crystalline Au nanoplates. Protein G formed on an ultraflat Au surface provides ideal background for dense and uniform immobilization of antibodies. The Au is highly stable in diverse biochemical environment and can immobilize antibodies easily through Au-S bonding, having been widely used for various biosensing applications. Especially, atomically smooth single-crystalline Au nanomaterials synthesized using chemical vapor transport (CVT) method are very suitable to fabricate reproducible sensitive sensors. As the C-reactive protein (CRP) is a nonspecific biomarker of inflammation and infection, it can be used as a predictive or prognostic marker for various cardiovascular diseases. Cys3-protein G immobilized uniformly on the Au nanoplate enable CRP antibody (anti-CRP) to be ordered in a correct orientation, making their binding capacity be maximized for CRP detection. Immobilization condition for the Cys3-protein G and anti-CRP on the Au nanoplate is optimized visually by AFM analysis. Au nanoparticle - Au nanoplate (NPs-on-Au nanoplate) assembly fabricated from sandwich immunoassay for CRP can reduce zero-signal extremely caused by nonspecific bindings, providing a distinct surface-enhanced Raman scattering (SERS) enhancement still in 10-18 M of CRP concentration. Moreover, the NP-on-Au nanoplate sensor shows an excellent selectivity against non-target proteins with high concentration. In addition, comparing with control experiments employing a Au film fabricated by e-beam assisted deposition and linker molecule, we validate clearly contribution of the Au nanoplate for the attomolar sensitive detection of CRP. We expect that the devised platform employing the complex of single-crystalline Au nanoplates and Cys3-protein G can be applied for detection of many other cancer biomarkers.

Keywords: Au nanoplate, biomarker, diagnostic sensor, protein G, SERS

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Authors: Mohammad Zafar-ul Islam

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The breeding programme of Houbara bustard was started in Saudi Arabia in 1986 to undertake the restoration of native species such as Houbara through a programme of re-introduction, involving the release of captive-bred birds in the wild. Two sites were selected for houbara re-introduction, i.e., Mahazat as-Sayd and Saja Umm Ar-Rimth protected areas in 1988 and 1998 respectively. Both the areas are fenced fairly level, sandy plain with a few rock outcrops. Captive bred houbara have been released in Mahazat since 1992 by NWRC and those birds have been successfully breeding since then. The nesting season of the houbara at Mahazat recorded from February to May and on an average 20-25 nests are located each year but no nesting recorded in Saja. Houbara are monitored using radio transmitters through aerial tracking technique and also a vehicle for terrestrial tracking. Total population of houbara in Mahazat is roughly estimated around 300-400 birds, using the following: N = n1+n2+n3+n4+n5 (n1 = released or wild-born, radio, regularly monitored/checked; n2 = radio tagged missing; n3 = wild born chicks not recorded; n4 = wild born chicks, recorded but not tagged; n5 = immigrants). However, in Saja only 4-7 individuals of houbara have been survived since 2001 because most of the birds are predated immediately after the release. The mean annual home was also calculated using Kernel and Convex polygons methods with Range VII software. The minimum density of houbara was also calculated. In order to know the houbara movement or their migration to other regions, two captive-reared male houbara that were released into the wild and one wild born female were fitted with Platform Transmitter Terminals (PTT). The home range shows that wild-born female has larger movement than two males. More areas need to be selected for reintroduction programme to establish the network of sites to provide easy access to move these birds and mingle with the wild houbara. Some potential sites have been proposed which require more surveys to check the habitat suitability.

Keywords: re-introduction, survival rate, home range, Saudi Arabia

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Authors: Lorico D. S. Lapitan Jr., Yihan Xu, Yuan Guo, Dejian Zhou

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The ability of ultrasensitive detection of specific genes and discrimination of single nucleotide polymorphisms is important for clinical diagnosis and biomedical research. Herein, we report the development of a new ultrasensitive approach for label-free DNA detection using magnetic nanoparticle (MNP) assisted rapid target capture/separation in combination with signal amplification using poly-enzyme tagged polymer nanobead. The sensor uses an MNP linked capture DNA and a biotin modified signal DNA to sandwich bind the target followed by ligation to provide high single-nucleotide polymorphism discrimination. Only the presence of a perfect match target DNA yields a covalent linkage between the capture and signal DNAs for subsequent conjugation of a neutravidin-modified horseradish peroxidase (HRP) enzyme through the strong biotin-nuetravidin interaction. This converts each captured DNA target into an HRP which can convert millions of copies of a non-fluorescent substrate (amplex red) to a highly fluorescent product (resorufin), for great signal amplification. The use of polymer nanobead each tagged with thousands of copies of HRPs as the signal amplifier greatly improves the signal amplification power, leading to greatly improved sensitivity. We show our biosensing approach can specifically detect an unlabeled DNA target down to 10 aM with a wide dynamic range of 5 orders of magnitude (from 0.001 fM to 100.0 fM). Furthermore, our approach has a high discrimination between a perfectly matched gene and its cancer-related single-base mismatch targets (SNPs): It can positively detect the perfect match DNA target even in the presence of 100 fold excess of co-existing SNPs. This sensing approach also works robustly in clinical relevant media (e.g. 10% human serum) and gives almost the same SNP discrimination ratio as that in clean buffers. Therefore, this ultrasensitive SNP biosensor appears to be well-suited for potential diagnostic applications of genetic diseases.

Keywords: DNA detection, polymer beads, signal amplification, single nucleotide polymorphisms

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Authors: Zelalem Fantahun

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Part-of-speech tagging is the process of assigning a part-of-speech or other lexical class marker to each word into naturally occurring text. Part-of-speech tagging is the most fundamental and basic task almost in all natural language processing. In natural language processing, the problem of providing large amount of manually annotated data is a knowledge acquisition bottleneck. Since, Amharic is one of under-resourced language, the availability of tagged corpus is the bottleneck problem for natural language processing especially for POS tagging. A promising direction to tackle this problem is to provide a system that does not require manually tagged data. In unsupervised learning, the learner is not provided with classifications. Unsupervised algorithms seek out similarity between pieces of data in order to determine whether they can be characterized as forming a group. This paper explicates the development of unsupervised part-of-speech tagger using K-Means clustering for Amharic language since large amount of data is produced in day-to-day activities. In the development of the tagger, the following procedures are followed. First, the unlabeled data (raw text) is divided into 10 folds and tokenization phase takes place; at this level, the raw text is chunked at sentence level and then into words. The second phase is feature extraction which includes word frequency, syntactic and morphological features of a word. The third phase is clustering. Among different clustering algorithms, K-means is selected and implemented in this study that brings group of similar words together. The fourth phase is mapping, which deals with looking at each cluster carefully and the most common tag is assigned to a group. This study finds out two features that are capable of distinguishing one part-of-speech from others these are morphological feature and positional information and show that it is possible to use unsupervised learning for Amharic POS tagging. In order to increase performance of the unsupervised part-of-speech tagger, there is a need to incorporate other features that are not included in this study, such as semantic related information. Finally, based on experimental result, the performance of the system achieves a maximum of 81% accuracy.

Keywords: POS tagging, Amharic, unsupervised learning, k-means

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Authors: Enerit Sacdanaku, Idriz Haxhiu

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This study was conducted in the area of Vlora Bay, Albania. Data about Sea Turtles Caretta caretta and Chelonia mydas, belonging to two periods of time (1984–1991; 2008–2014) are given. All data gathered were analyzed using recent methodologies. For all turtles captured (as by catch), the Curve Carapace Length (CCL) and Curved Carapace Width (CCW) were measured. These data were statistically analyzed, where the mean was 67.11 cm for CCL and 57.57 cm for CCW of all individuals studied (n=13). All untagged individuals of marine turtles were tagged using metallic tags (Stockbrand’s titanium tag) with an Albanian address. Sex was determined and resulted that 45.4% of individuals were females, 27.3% males and 27.3% juveniles. All turtles were studied for the presence of the epibionts. The area of Vlora Bay is used from marine turtles (Caretta caretta) as a migratory corridor to pass from the Mediterranean to the northern part of the Adriatic Sea.

Keywords: Caretta caretta, Chelonia mydas, CCL, CCW, tagging, Vlora Bay

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Authors: Minami Ito, Akihiro Iijima

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On March 11, 2011, the Great East Japan Earthquake occurred off the coast of Sanriku, Japan. It is important to build a sustainable society through the reconstruction process rather than simply restoring the infrastructure. To compare the goals of reconstruction plans of quake-stricken municipalities, Japanese language morphological analysis was performed by using text mining techniques. Frequently-used nouns were sorted into four main categories of “life”, “disaster prevention”, “economy”, and “harmony with environment”. Because Soma City is affected by nuclear accident, sentences tagged to “harmony with environment” tended to be frequent compared to the other municipalities. Results from cluster analysis and principle component analysis clearly indicated that the local government reinforces the efforts to reduce risks from radiation exposure as a top priority.

Keywords: eco-friendly reconstruction, harmony with environment, decontamination, nuclear disaster

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Authors: A. R. Momeninezhad

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Erosion of Slurry Pumps in Related Industries, is one of the major costs in their production process. Many factories in extractive industries try to find ways to diminish this cost. In this paper, we consider the flow pattern modifications by geometric variations made of numerical simulation of flow inside pump casing, which is one of the most important parts analyzed for erosion. The mentioned pump is a cyclone centrifugal slurry pump, which is operating in Sarcheshmeh Copper Industries in Kerman-Iran, named and tagged as HM600 cyclone pump. Simulation shows many improvements in local wear information and situations for better and more qualified design of casing shape and impeller position, before and after geometric corrections. By theory of liquid-solid two-phase flow, the local wear defeats are analyzed and omitted.

Keywords: flow pattern, slurry pump, simulation, wear

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Authors: Yakup Ulusu, Faruk Ozel, Numan Eczacioglu, Abdurrahman Ozen, Sabriye Acikgoz

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GFP discovered in the mid-1970s, has been used as a marker after replicated genetic study by scientists. In biotechnology, cell, molecular biology, the GFP gene is frequently used as a reporter of expression. In modified forms, it has been used to make biosensors. Many animals have been created that express GFP as an evidence that a gene can be expressed throughout a given organism. Proteins labeled with GFP identified locations are determined. And so, cell connections can be monitored, gene expression can be reported, protein-protein interactions can be observed and signals that create events can be detected. Additionally, monitoring GFP is noninvasive; it can be detected by under UV-light because of simply generating fluorescence. Moreover, GFP is a relatively small and inert molecule, that does not seem to treat any biological processes of interest. The synthesis of GFP has some steps like, to construct the plasmid system, transformation in E. coli, production and purification of protein. GFP carrying plasmid vector pBAD–GFPuv was digested using two different restriction endonuclease enzymes (NheI and Eco RI) and DNA fragment of GFP was gel purified before cloning. The GFP-encoding DNA fragment was ligated into pET28a plasmid using NheI and Eco RI restriction sites. The final plasmid was named pETGFP and DNA sequencing of this plasmid indicated that the hexa histidine-tagged GFP was correctly inserted. Histidine-tagged GFP was expressed in an Escherichia coli BL21 DE3 (pLysE) strain. The strain was transformed with pETGFP plasmid and grown on LuiraBertoni (LB) plates with kanamycin and chloramphenicol selection. E. coli cells were grown up to an optical density (OD 600) of 0.8 and induced by the addition of a final concentration of 1mM isopropyl-thiogalactopyranoside (IPTG) and then grown for additional 4 h. The amino-terminal hexa-histidine-tag facilitated purification of the GFP by using a His Bind affinity chromatography resin (Novagen). Purity of GFP protein was analyzed by a 12 % sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). The concentration of protein was determined by UV absorption at 280 nm (Varian Cary 50 Scan UV/VIS spectrophotometer). Synthesis of GFP-Polymer composite nanofibers was produced by using GFP solution (10mg/mL) and polymer precursor Polyvinylpyrrolidone, (PVP, Mw=1300000) as starting materials and template, respectively. For the fabrication of nanofibers with the different fiber diameter; a sol–gel solution comprising of 0.40, 0.60 and 0.80 g PVP (depending upon the desired fiber diameter) and 100 mg GFP in 10 mL water: ethanol (3:2) mixtures were prepared and then the solution was covered on collecting plate via electro spinning at 10 kV with a feed-rate of 0.25 mL h-1 using Spellman electro spinning system. Results show that GFP-based nano-fiber can be used plenty of biomedical applications such as bio-imaging, bio-mechanic, bio-material and tissue engineering.

Keywords: biomaterial, GFP, nano-fibers, protein expression

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Authors: Nessrine Jebari, Elisabeth Dufour-Gergam, Mehdi Ammar

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This work outlines the development and characterization of a microfluidic immunosensor, designed for real-time monitoring of proinflammatory pathologies, within the domain of biomedical diagnostics. The key focus of this research is the microfabrication protocol developed in a cleanroom environment, emphasizing the intricate processes employed in the fabrication of this sophisticated biosensor. The principal contribution of this project is grounded in the application of COMSOL Multiphysics for detailed three-dimensional simulations, which are instrumental in the development of a patch-like device designed for non-invasive monitoring of biomarkers in sweat. The device integrates a distinctive combination of magnetofluidic manipulation and capacitive sensing approaches, thereby enhancing the standard in biomarker detection and quantification. A pivotal aspect of our device is its use of biomarker-tagged magnetic nanoparticles (MNPs), significantly enhancing sensitivity and multifunctionality. The immunosensor comprises two main units: the primary unit, featuring an array of serial microcoils for optimal MNP trapping and microfluidic mixing, and the secondary unit, a layered structure with a planar microcoil and two copper electrodes. This configuration forms a capacitor integral to the capacitive sensing capabilities of the device, allowing for precise quantification of biomarker-tagged MNPs. The microfabrication process, executed in a controlled cleanroom environment, involved the precise layering and structuring of microcoils and electrodes, along with the integration of a microfluidic platform. This meticulous process ensures high reproducibility and accuracy, critical for a reliable diagnostic tool. Experimental results exhibit the immunosensor's promoting sensing capabilities, with a sensitivity range of 60% to 75% at 70% MNP occupancy in the detection zone. This performance underscores the device's potential in overcoming the limitations of surface biochemical functionalization, a challenge in conventional biosensors. Beyond monitoring proinflammatory pathologies, the immunosensor's versatility extends to detecting a wide range of pathogens, including bacteria. Its compatibility with complementary screening techniques allows for the identification of multiple biomarkers, enhancing its utility in both clinical and research settings. Conclusion: In conclusion, the development of this microfluidic immunosensor marks a significant result in biomedical engineering and diagnostic technology. The microfabrication process, executed in a cleanroom environment, is crucial to the device's distinctive design and its multifunctional attributes. This technology not only progresses the realm of biomedical diagnostics but also paves the way for further exploration and development within the discipline. Its applications extend from early disease detection to the monitoring of treatment effectiveness.

Keywords: COMSOL multiphysics 3D simulation, microfluidic immunosensor, magnetofluidic manipulation, magnetic nanoparticle trapping, laboratory-on-patch technology

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Authors: Olusola Joseph Adesina, Adebayo Adeyinka Salako

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The study adopted a descriptive research design. Two hundred (200) in-school adolescents were purposely selected in Afijio Local Government Area of Oyo State. Two hypotheses were also raised to pilot the study. The researchers developed an instrument which was validated by psychological experts, the instrument tagged counseling psychology and sexual risk behavior questionnaire (CPSRBQ)(r = 0.78). The results were analysed using t-test at 0.05 level of significance. The result showed that there is a significant relationship between counseling psychology and sexual risk behavior of in-school adolescents. It was also noticed that there is a significant difference in the sexual risk behavior of male and female adolescents. Based on the findings, it was recommended that more counselors are still needed in Nigeria schools. There is need for restructuring Nigeria Curriculum most especially on sex education and related diseases. Lastly, adolescents should be more exposed to seminars on HIV/AIDS, sex education enlightenment programmes and marital counseling.

Keywords: counseling psychology, sexual behavior, risk and adolescent, cognitive sciences

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Authors: Sham Kidane, Fitsum Gaim, Ibrahim Abdella, Sirak Asmerom, Yoel Ghebrihiwot, Simon Mulugeta, Natnael Ambassager

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The lack of annotated datasets is a bottleneck to the progress of NLP in low-resourced languages. The work presented here consists of large-scale annotated datasets and models for the named entity recognition (NER) system for the Tigrinya language. Our manually constructed corpus comprises over 340K words tagged for NER, with over 118K of the tokens also having parts-of-speech (POS) tags, annotated with 12 distinct classes of entities, represented using several types of tagging schemes. We conducted extensive experiments covering convolutional neural networks and transformer models; the highest performance achieved is 88.8% weighted F1-score. These results are especially noteworthy given the unique challenges posed by Tigrinya’s distinct grammatical structure and complex word morphologies. The system can be an essential building block for the advancement of NLP systems in Tigrinya and other related low-resourced languages and serve as a bridge for cross-referencing against higher-resourced languages.

Keywords: Tigrinya NER corpus, TiBERT, TiRoBERTa, BiLSTM-CRF

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Authors: DEVASHISH KHULBE, MANU PATHAK

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The rise in popularity of major social media platforms have enabled people to share photos and textual information about their daily life. One of the popular topics about which information is shared is food. Since a lot of media about food are attributed to particular locations and restaurants, information like spatio-temporal popularity of various cuisines can be analyzed. Tracking the popularity of food types and retail locations across space and time can also be useful for business owners and restaurant investors. In this work, we present an approach using off-the shelf machine learning techniques to identify trends and popularity of cuisine types in an area using geo-tagged data from social media, Google images and Yelp. After adjusting for time, we use the Kernel Density Estimation to get hot spots across the location and model the dependencies among food cuisines popularity using Bayesian Networks. We consider the Manhattan borough of New York City as the location for our analyses but the approach can be used for any area with social media data and information about retail businesses.

Keywords: Web Mining, Geographic Information Systems, Business popularity, Spatial Data Analyses

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Authors: Omoniyi A. Olubunmi, Omoniyi Olayide M.

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This study was designed to assess the attitudes of Nigerian university teachers by their students in terms of teachers’ attitude to work, teaching and students. The study was a survey, and made use of the researcher’s designed questionnaire tagged Students’ Assessment Teachers Inventory (SATI), comprising 20 items, was used to collect data. The respondents were 300 students which were randomly selected from three universities in Ondo State. The SATI elicited information on different aspects of teachers’ attitude to work, teaching and students. The study was guided by two hypotheses. Data collected were analyzed using Pearson-r. The result showed that there was a significant relationship between teachers’ attitude to work (r = 0.343, p<0.01), teaching (r = 0.594, p<0.01) and students (r = 0.487, p<0.01). The study concluded that teachers’ attitudes to teaching profession in higher institutions in Ondo State were not favorable and this could be improved through capacity building for effective pedagogical skills, conducive environment, well equipped libraries and laboratories, and provision of incentives for university teachers.

Keywords: capacity building, pedagogical skills, teachers’ attitude, students’ assessment

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Authors: Khadijat T. Bamigbade, Olufade F. W. Onifade

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The field of automatic facial expression analysis has been an active research area in the last two decades. Its vast applicability in various domains has drawn so much attention into developing techniques and dataset that mirror real life scenarios. Many techniques such as Local Binary Patterns and its variants (CLBP, LBP-TOP) and lately, deep learning techniques, have been used for facial expression recognition. However, the problem of occlusion has not been sufficiently handled, making their results not applicable in real life situations. This paper develops a simple, yet highly efficient method tagged Local Binary Pattern-Histogram of Gradient (LBP-HOG) with occlusion detection in face image, using a multi-class SVM for Action Unit and in turn expression recognition. Our method was evaluated on three publicly available datasets which are JAFFE, CK, SFEW. Experimental results showed that our approach performed considerably well when compared with state-of-the-art algorithms and gave insight to occlusion detection as a key step to handling expression in wild.

Keywords: automatic facial expression analysis, local binary pattern, LBP-HOG, occlusion detection

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Authors: Iñaki Iturria, Pasquale Russo, Montserrat Nacher-Vázquez, Giuseppe Spano, Paloma López, Miguel Angel Pardo

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Introduction: It is known that some Lactic Acid Bacteria (LAB) present an interesting probiotic effect. Probiotic bacteria stimulate host resistance to microbial pathogens and thereby aid in immune response, and modulate the host's immune responses to antigens with a potential to down-regulate hypersensitivity reactions. Therefore, probiotic therapy is valuable against intestinal infections and may be beneficial in the treatment of Inflammatory Bowel Disease (IBD). Several in vitro tests are available to evaluate the probiotic potential of a LAB strain. However, an in vivo model is required to understand the interaction between the host immune system and the bacteria. During the last few years, zebrafish (Danio rerio) has gained interest as a promising vertebrate model in this field. This organism has been extensively used to study the interaction between the host and the microbiota, as well as the host immune response under several microbial infections. In this work, we report on the use of the zebrafish model to investigate in vivo the colonizing ability and the immunomodulatory effect of probiotic LAB. Methods: Lactobacillus strains belonging to different LAB species were fluorescently tagged and used to colonize germ-free zebrafish larvae gastrointestinal tract (GIT). Some of the strains had a well-documented probiotic effect (L. acidophilus LA5); while others presented an exopolysaccharide (EPS) producing phenotype, thus allowing evaluating the influence of EPS in the colonization and immunomodulatory effect. Bacteria colonization was monitored for 72 h by direct observation in real time using fluorescent microscopy. CFU count per larva was also evaluated at different times. The immunomodulatory effect was assessed analysing the differential expression of several innate immune system genes (MyD88, NF-κB, Tlr4, Il1β and Il10) by qRT- PCR. The anti-inflammatory effect was evaluated using a chemical enterocolitis zebrafish model. The protective effect against a pathogen was also studied. To that end, a challenge test was developed using a fluorescently tagged pathogen (Vibrio anguillarum-GFP+). The progression of the infection was monitored up to 3 days using a fluorescent stereomicroscope. Mortality rates and CFU counts were also registered. Results and conclusions: Larvae exposed to EPS-producing bacteria showed a higher fluorescence and CFU count than those colonized with no-EPS phenotype LAB. In the same way, qRT-PCR results revealed an immunomodulatory effect on the host after the administration of the strains with probiotic activity. A downregulation of proinflammatory cytoquines as well as other cellular mediators of inflammation was observed. The anti-inflammatory effect was found to be particularly marked following exposure to LA% strain, as well as EPS producing strains. Furthermore, the challenge test revealed a protective effect of probiotic administration. As a matter of fact, larvae fed with probiotics showed a decrease in the mortality rate ranging from 20 to 35%. Discussion: In this work, we developed a promising model, based on the use of gnotobiotic zebrafish coupled with a bacterial fluorescent tagging in order to evaluate the probiotic potential of different LAB strains. We have successfully used this system to monitor in real time the colonization and persistence of exogenous LAB within the gut of zebrafish larvae, to evaluate their immunomodulatory effect and for in vivo competition assays. This approach could bring further insights into the complex microbial-host interactions at intestinal level.

Keywords: gnotobiotic, immune system, lactic acid bacteria, probiotics, zebrafish

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Authors: J. Carlos Kuri, Varun Vij, Raymond J. Turner, Orly Yadid-Pecht

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Celiac disease (CD) is an immune system disorder that is triggered by ingesting gluten. As a gluten-free (GF) diet has become a concern of many people for health reasons, a gold standard had to be nominated. Enzyme-linked immunosorbent assay (ELISA) has taken the seat of this role. However, multiple limitations were discovered, and with that, the desire for an alternative method now exists. Nucleic acid-based aptamers have become of great interest due to their selectivity, specificity, simplicity, and rapid-testing advantages. However, fluorescence-based aptasensors have been tagged as unstable, but lifespan details are rarely stated. In this work, the lifespan stability of a fluorescence-based aptasensor is shown over an 8-week-long study displaying the accuracy of the sensor and false negatives. This study follows 22 different samples, including GF and gluten-rich (GR) and soy sauce products, off-the-shelf products, and reference material from laboratories, giving a total of 836 tests. The analysis shows an accuracy of correctly classifying GF and GR products of 96.30% and 100%, respectively when the protocol is augmented with molecular sieves. The overall accuracy remains around 94% within the first four weeks and then decays to 63%.

Keywords: aptasensor, PEG, rGO, FAM, RM, ELISA

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Authors: Vishal Kumar, Soumitra Satapathi

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Föster Resonance Energy Transfer (FRET) is a powerful technique used to probe close-range molecular interactions. Physically, the FRET phenomenon manifests as a dipole–dipole interaction between closely juxtaposed fluorescent molecules (10–100 Å). Our effort is to employ this FRET technique to make a prototype device for highly sensitive detection of environment pollutant. Among the most common environmental pollutants, nitroaromatic compounds (NACs) are of particular interest because of their durability and toxicity. That’s why, sensitive and selective detection of small amounts of nitroaromatic explosives, in particular, 2,4,6-trinitrophenol (TNP), 2,4-dinitrotoluene (DNT) and 2,4,6-trinitrotoluene (TNT) has been a critical challenge due to the increasing threat of explosive-based terrorism and the need of environmental monitoring of drinking and waste water. In addition, the excessive utilization of TNP in several other areas such as burn ointment, pesticides, glass and the leather industry resulted in environmental accumulation, and is eventually contaminating the soil and aquatic systems. To the date, high number of elegant methods, including fluorimetry, gas chromatography, mass, ion-mobility and Raman spectrometry have been successfully applied for explosive detection. Among these efforts, fluorescence-quenching methods based on the mechanism of FRET show good assembly flexibility, high selectivity and sensitivity. Here, we report a FRET-based sensor system for the highly selective detection of NACs, such as TNP, DNT and TNT. The sensor system is composed of a copolymer Poly [(N,N-dimethylacrylamide)-co-(Boc-Trp-EMA)] (RP) bearing tryptophan derivative in the side chain as donor and dansyl tagged copolymer P(MMA-co-Dansyl-Ala-HEMA) (DCP) as an acceptor. Initially, the inherent fluorescence of RP copolymer is quenched by non-radiative energy transfer to DCP which only happens once the two molecules are within Förster critical distance (R0). The excellent spectral overlap (Jλ= 6.08×10¹⁴ nm⁴M⁻¹cm⁻¹) between donors’ (RP) emission profile and acceptors’ (DCP) absorption profile makes them an exciting and efficient FRET pair i.e. further confirmed by the high rate of energy transfer from RP to DCP i.e. 0.87 ns⁻¹ and lifetime measurement by time correlated single photon counting (TCSPC) to validate the 64% FRET efficiency. This FRET pair exhibited a specific fluorescence response to NACs such as DNT, TNT and TNP with 5.4, 2.3 and 0.4 µM LODs, respectively. The detection of NACs occurs with high sensitivity by photoluminescence quenching of FRET signal induced by photo-induced electron transfer (PET) from electron-rich FRET pair to electron-deficient NAC molecules. The estimated stern-volmer constant (KSV) values for DNT, TNT and TNP are 6.9 × 10³, 7.0 × 10³ and 1.6 × 104 M⁻¹, respectively. The mechanistic details of molecular interactions are established by time-resolved fluorescence, steady-state fluorescence and absorption spectroscopy confirmed that the sensing process is of mixed type, i.e. both dynamic and static quenching as lifetime of FRET system (0.73 ns) is reduced to 0.55, 0.57 and 0.61 ns DNT, TNT and TNP, respectively. In summary, the simplicity and sensitivity of this novel FRET sensor opens up the possibility of designing optical sensor of various NACs in one single platform for developing multimodal sensor for environmental monitoring and future field based study.

Keywords: FRET, nitroaromatic, stern-Volmer constant, tryptophan and dansyl tagged copolymer

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Authors: A. Ballard

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The ability to track goods and products en route in the delivery system, in the warehouse, and on the top floor is a huge advantage to shippers and retailers. Recently the emergence of radio frequency identification (RFID) technology has enabled this better than ever before. However, a significant problem exists in that RFID technology depends on the quality of the information stored for each tagged product. Because of the profusion of names for colours, it is very difficult to ascertain that stored values are recognised by all users who view the product visually. This paper reports the findings of a study in which 50 consumers and 50 logistics workers were shown colour swatches and asked to choose the name of the colour from a multiple choice list. They were then asked to match consumer products, including toasters, jumpers, and toothbrushes, with the identifying inventory information available for each one. The findings show that the ability to match colours was significantly stronger with the color swatches than with the consumer products and that while logistics professionals made more frequent correct identification than the consumers, their results were still unsatisfactorily low. Based on these findings, a proposed universal model of colour identification numbers has been developed.

Keywords: consumer preferences, supply chain logistics, radio frequency identification, RFID, colour preference

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Authors: J. Bernasovská, R. Lohajová Behulová, E. Petrejčiková, I. Boroňová, I. Bernasovský

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Y chromosome microdeletions are the most common genetic cause of male infertility and screening for these microdeletions in azoospermic or severely oligospermic men is now standard practice. Analysis of the Y chromosome in men with azoospermia or severe oligozoospermia has resulted in the identification of three regions in the euchromatic part of the long arm of the human Y chromosome (Yq11) that are frequently deleted in men with otherwise unexplained spermatogenic failure. PCR analysis of microdeletions in the AZFa, AZFb and AZFc regions of the human Y chromosome is an important screening tool. The aim of this study was to analyse the type of microdeletions in men with fertility disorders in Slovakia. We evaluated 227 patients with azoospermia and with normal karyotype. All patient samples were analyzed cytogenetically. For PCR amplification of sequence-tagged sites (STS) of the AZFa, AZFb and AZFc regions of the Y chromosome was used Devyser AZF set. Fluorescently labeled primers for all markers in one multiplex PCR reaction were used and for automated visualization and identification of the STS markers we used genetic analyzer ABi 3500xl (Life Technologies). We reported 13 cases of deletions in the AZF region 5,73%. Particular types of deletions were recorded in each region AZFa,b,c .The presence of microdeletions in the AZFc region was the most frequent. The study confirmed that percentage of microdeletions in the AZF region is low in Slovak azoospermic patients, but important from a prognostic view.

Keywords: AZF, male infertility, microdeletions, Y chromosome

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Authors: Shamm Kidane, Ibrahim Abdella, Fitsum Gaim, Simon Mulugeta, Sirak Asmerom, Natnael Ambasager, Yoel Ghebrihiwot

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The call for natural language processing (NLP) systems for low-resource languages has become more apparent than ever in the past few years, with the arduous challenges still present in preparing such systems. This paper presents an improved dataset version of the Nagaoka Tigrinya Corpus for Parts-of-Speech (POS) classification system in the Tigrinya language. The size of the initial Nagaoka dataset was incremented, totaling the new tagged corpus to 118K tokens, which comprised the 12 basic POS annotations used previously. The additional content was also annotated manually in a stringent manner, followed similar rules to the former dataset and was formatted in CONLL format. The system made use of the novel approach in NLP tasks and use of the monolingually pre-trained TiELECTRA, TiBERT and TiRoBERTa transformer models. The highest achieved score is an impressive weighted F1-score of 94.2%, which surpassed the previous systems by a significant measure. The system will prove useful in the progress of NLP-related tasks for Tigrinya and similarly related low-resource languages with room for cross-referencing higher-resource languages.

Keywords: Tigrinya POS corpus, TiBERT, TiRoBERTa, conditional random fields

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Authors: Achraf Al Faraj, Asma Sultana Shaik, Baraa Al Sayed

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Specific and effective targeting of drug delivery systems (DDS) to cancerous sites remains a major challenge for a better diagnostic and therapy. Recently, SWCNTs with their unique physicochemical properties and the ability to cross the cell membrane show promising in the biomedical field. The purpose of this study was first to develop a biocompatible iron oxide tagged SWCNTs as diagnostic nanoprobes to allow their noninvasive detection using MRI and their preferential targeting in a breast cancer murine model by placing an optimized flexible magnet over the tumor site. Magnetic targeting was associated to specific antibody-conjugated SWCNTs active targeting. The therapeutic efficacy of doxorubicin-conjugated SWCNTs was assessed, and the superiority of diffusion-weighted (DW-) MRI as sensitive imaging biomarker was investigated. Short Polyvinylpyrrolidone (PVP) stabilized water soluble SWCNTs were first developed, tagged with iron oxide nanoparticles and conjugated with Endoglin/CD105 monoclonal antibodies. They were then conjugated with doxorubicin drugs. SWCNTs conjugates were extensively characterized using TEM, UV-Vis spectrophotometer, dynamic light scattering (DLS) zeta potential analysis and electron spin resonance (ESR) spectroscopy. Their MR relaxivities (i.e. r1 and r2*) were measured at 4.7T and their iron content and metal impurities quantified using ICP-MS. SWCNTs biocompatibility and drug efficacy were then evaluated both in vitro and in vivo using a set of immunological assays. Luciferase enhanced bioluminescence 4T1 mouse mammary tumor cells (4T1-Luc2) were injected into the right inguinal mammary fat pad of Balb/c mice. Tumor bearing mice received either free doxorubicin (DOX) drug or SWCNTs with or without either DOX or iron oxide nanoparticles. A multi-pole 10x10mm high-energy flexible magnet was maintained over the tumor site during 2 hours post-injections and their properties and polarity were optimized to allow enhanced magnetic targeting of SWCNTs toward the primary tumor site. Tumor volume was quantified during the follow-up investigation study using a fast spin echo MRI sequence. In order to detect the homing of SWCNTs to the main tumor site, susceptibility-weighted multi-gradient echo (MGE) sequence was used to generate T2* maps. Apparent diffusion coefficient (ADC) measurements were also performed as a sensitive imaging biomarker providing early and better assessment of disease treatment. At several times post-SWCNT injection, histological analysis were performed on tumor extracts and iron-loaded SWCNT were quantified using ICP-MS in tumor sites, liver, spleen, kidneys, and lung. The optimized multi-poles magnet revealed an enhanced targeting of magnetic SWCNTs to the primary tumor site, which was found to be much higher than the active targeting achieved using antibody-conjugated SWCNTs. Iron-loading allowed their sensitive noninvasive tracking after intravenous administration using MRI. The active targeting of doxorubicin through magnetic antibody-conjugated SWCNTs nanoprobes was found to considerably decrease the primary tumor site and may have inhibited the development of metastasis in the tumor-bearing mice lung. ADC measurements in DW-MRI were found to significantly increase in a time-dependent manner after the injection of DOX-conjugated SWCNTs complexes.

Keywords: single-walled carbon nanotubes, nanomedicine, magnetic resonance imaging, cancer diagnosis and therapy

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Authors: Amit Kumar Jain, Surbhi Jain

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The urban freight in a city constitutes 10 to 18 per cent of all city road traffic, and 40 per cent of air pollution and noise emissions, are directly related to commercial transport. The policy measures implemented by urban planners have sought to restrict rather than assist goods-vehicle operations. This approach has temporarily controlled the urban transport demand during peak hours of traffic but has not effectively solved transport congestion. The solution discussed in the paper envisages the development of a comprehensive network of Urban Freight Stations (UFS) connected through underground conveyor belts in the city in line with baggage segregation and distribution in any of the major airports. The transportation of freight shall be done in standard size containers/cars through rail borne carts. The freight can be despatched or received from any of the UFS. Once freight is booked for a destination from any of the UFS, it would be stuffed in the container and digitally tagged for the destination. The container would reach the destination UFS through a network of rail borne carts. The container would be de-stuffed at the destination UFS and sent for further delivery, or the consignee may be asked to collect the consignment from urban freight station. The obvious benefits would be decongestion of roads, reduction in air and noise pollution, saving in manpower used for freight transportation.

Keywords: congestion, urban freight, intelligent transport system, pollution

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Authors: Christian Nwaoha

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This study investigates taboo expressions and classifications in Igbo discourse, their socio-cultural factors affecting their usage. The study classifies Linguistic taboo expressions by their discourse into five categories: morality-related taboo, veneration-related, decorum-related, religion-related and fear-related taboo expressions. This study argues that while religion-related and decorum-related taboos are unmentioned and have no euphemistic synonyms is because they are closely tied to various Igbo deities and objects, while morality, veneration, and fear-related have permissible alternatives. A descriptive research design was adopted and the data collection was by questionnaire and oral interview. The result of the research proves that aside of the categories of taboos in Igbo, socially, the styles of discourse have some levels of gender, age and class-connected taboos, which for instance, in gender-connected taboos, women in Igbo are forbidden to use style of discourse that are connected with genital organs in social gathering comprising men and women. The same has to do with class-connected where much younger men can use some certain expressions that are taboo, but in much older men gathering such expressions would be tagged forbidden in the context. The study further reveals that there are occasions in which these taboos can be used with reasons. The research concludes that using these taboos in literary text can enhance clear understanding of Igbo taboos to the users and learners of Igbo language.

Keywords: taboo expressions, classifications, Igbo, socio-cultural factors, discourse

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Authors: Mitakshara Sharma, Sonal Sharma

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Vesiculobullous diseases are heterogeneous group of dermatological disorders with protean manifestations. The most important technique for the patients with vesiculobullous diseases is conventional histopathology and confirmatory tests like direct immunofluorescence (DIF) and indirect immunofluorescence (IIF). DIF has been used for decades to investigate pathophysiology and in the diagnosis. It detects molecules such as immunoglobulins and complement components. It is done on the perilesional skin. Diagnosis of DIF test depends on features like primary site of the immune deposits, class of immunoglobulin, number of immune deposits and deposition at other sites. The aim of the study is to correlate DIF with clinical and histopathological findings and to analyze the utility of DIF in the diagnosis of these disorders. It is a retrospective descriptive study conducted for 2 years from 2015 to 2017 in Department of Pathology, GTB Hospital on perilesional punch biopsies of vesiculobullous lesions. Biopsies were sent in Michael’s medium. The specimens were washed, frozen and incubated with fluorescein isothiocyanate (FITC) tagged antihuman antibodies IgA, IgG, IgM, C3 & F and were viewed under fluorescent microscope. Out of 401 skin biopsies submitted for DIF, 285 were vesiculobullous diseases, in which the most common was Pemphigus vulgaris (34%) followed by Bullous pemphigoid (21.5%), Dermatitis herpetiformis (16%), Pemphigus foliaceus (11.9%), Linear IgA disease (11.9%), Epidermolysisbullosa (2.39%) and Pemphigus herpetiformis (1.7%). We will be presenting the DIF findings in the all these vesiculobullous diseases. DIF in conjugation with histopathology gives the best diagnostic yield in these lesions. It also helps in the diagnosis whenever there is a clinical and histopathological overlap.

Keywords: antibodies, direct immunofluorescence, pemphigus, vesiculobullous

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