Search results for: cytotoxicity test

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

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

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

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Authors: Hajar Hosseini Khorami

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Porphyrins are organic, aromatic compounds found in heme, cytochrome, cobalamin, chlorophyll , and many other natural products with essential roles in biological processes that their cationic forms have been used as groups of favorable non-viral vectors recently. Cationic porphyrins are self-chromogenic reagents with a high capacity for modifications, great interaction with DNA and protection of DNA from nuclease during delivery of it into a cell with low toxicity. In order to have high efficient gene transfection into the cell while causing low toxicity, genetically manipulations of the non-viral vector, cationic porphyrin, would be useful. In this study newly modified cationic porphyrin derivative, 5, 10-dihexyl-15, 20bis (N-methyl-4-pyridyl) porphyrin was applied. Cytotoxicity of synthesized cationic porphyrin on Chinese Hamster Ovarian (CHO) cells was evaluated by using MTT assay. This cationic derivative is dose-dependent, with low cytotoxicity at the ranges from 100 μM to 0.01μM. It was uptake by cells at high concentration. Using direct non-viral gene transfection method and different concentration of cationic porphyrin were tested on transfection of CHO cells by applying derived transfection reagent with X-tremeGENE HP DNA as a positive control. However, no transfection observed by porphyrin derivative and the parameters tested except for positive control. Results of this study suggested that applying different protocol, and also trying other concentration of cationic porphyrins and DNA for forming a strong complex would increase the possibility of efficient gene transfection by using cationic porphyrins.

Keywords: cationic porphyrins, gene delivery, non-viral vectors, transfection reagents

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Authors: Adriano Bessa Albuquerque, Francisco Jose Barreto Nunes

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Software vulnerabilities are increasing and not only impact services and processes availability as well as information confidentiality, integrity and privacy, but also cause changes that interfere in the development process. Security test could be a solution to reduce vulnerabilities. However, the variety of test techniques with the lack of real case studies of applying tests focusing on software development life cycle compromise its effective use. This paper offers an overview of how a Systematic Mapping Study (MS) about security verification, validation and test (VVT) was performed, besides presenting general results about this study.

Keywords: software test, software security verification validation and test, security test institutionalization, systematic mapping study

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Authors: Francisco J. Cedano, Laura M. Pinzón, Camila I. Castro, Felipe Salcedo, Juan P. Casas, Juan C. Briceño

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Complex fractures located in articular surfaces are challenging to treat and their reduction with conventional treatments could compromise the functionality of the affected limb. An adhesive material to treat those fractures is desirable for orthopedic surgeons. This adhesive must be biocompatible and have a high adhesion to bone surface in an aqueous environment. The proposed adhesive is based on chitosan, given its adhesive and biocompatibility properties. Chitosan is mixed with calcium carbonate and hydroxyapatite, which contribute to structural support and a gel like behavior, and glutaraldehyde is used as a cross-linking agent to keep the adhesive mechanical performance in aqueous environment. This work aims to evaluate the rheological, adhesion strength and biocompatibility properties of the proposed adhesive using in vitro tests. The gelification process of the adhesive was monitored by oscillatory rheometry in an ARG-2 TA Instruments rheometer, using a parallel plate geometry of 22 mm and a gap of 1 mm. Time sweep experiments were conducted at 1 Hz frequency, 1% strain and 37°C from 0 to 2400 s. Adhesion strength is measured using a butt joint test with bovine cancellous bone fragments as substrates. The test is conducted at 5 min, 20min and 24 hours after curing the adhesive under water at 37°C. Biocompatibility is evaluated by a cytotoxicity test in a fibroblast cell culture using MTT assay and SEM. Rheological results concluded that the average gelification time of the adhesive is 820±107 s, also it reaches storage modulus magnitudes up to 106 Pa; The adhesive show solid-like behavior. Butt joint test showed 28.6 ± 9.2 kPa of tensile bond strength for the adhesive cured for 24 hours. Also there was no significant difference in adhesion strength between 20 minutes and 24 hours. MTT showed 70 ± 23 % of active cells at sixth day of culture, this percentage is estimated respect to a positive control (only cells with culture medium and bovine serum). High vacuum SEM observation permitted to localize and study the morphology of fibroblasts presented in the adhesive. All captured fibroblasts presented in SEM typical flatted structure with filopodia growth attached to adhesive surface. This project reports an adhesive based on chitosan that is biocompatible due to high active cells presented in MTT test and these results were correlated using SEM. Also, it has adhesion properties in conditions that model the clinical application, and the adhesion strength do not decrease between 5 minutes and 24 hours.

Keywords: bioadhesive, bone adhesive, calcium carbonate, chitosan, hydroxyapatite, glutaraldehyde

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Authors: Seyed Ebrahim Hosseini, Shahbaz Pervez

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As people go through aging, maintenance and improvement of the quality of life become more important for them. To know the role of smartphone technology and communications applications on quality of life, a sample group of old people living in Dar-Iran was selected for a quasi-experimental study. The participants were registered senior inhabitants from public health centers in Dar, Iran in 2022. The number of participants was 39. Participants were randomly categorized into intervention and control groups. A validated Persian version of the Control, Autonomy, Self-realisation, Pleasure scale (CASP-19) scale questionnaire was provided for them which answers were used for the quality of life assessment. The paired t-test between pre-and post-test (t= -8.45, p<.00), post-and follow-up-test (t= -2.12, p=.01), and pre-test and the follow-up test (t= -8.27, p<.00) in the intervention group revealed a considerable mean difference. Based on the results of paired t-test for the control group, this was not significant between pre-test and post-test (t= 1.26, p=.15), post-test and follow-up test (t= .33, p=.67) and pre-test and follow-up test (t= 1.85, p=.08) for quality of life. Considering the educational training associated with it, this study aimed at helping families and aging field practitioners to understand the essentiality of modern communication technology teaching in promoting a greater life quality of the elderly’s community.

Keywords: Iranian elderly, quality of life, smartphone, social applications, CASP-19

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Authors: A. Zuchowska, A. Buta, M. Mazurkiewicz-Pawlicka, A. Malolepszy, L. Stobinski, Z. Brzozka

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In recent years, graphene-based materials are finding more and more applications in biological science. As a thin, tough, transparent and chemically resistant materials, they appear to be a very good material for the production of implants and biosensors. Interest in graphene derivatives also resulted at the beginning of research about the possibility of their application in cancer therapy. Currently, the analysis of their potential use in photothermal therapy and as a drug carrier is mostly performed. Moreover, the direct anticancer properties of graphene-based materials are also tested. Nowadays, cytotoxic studies are conducted on in vitro cell culture in standard culture vessels (macroscale). However, in this type of cell culture, the cells grow on the synthetic surface in static conditions. For this reason, cell culture in macroscale does not reflect in vivo environment. The microfluidic systems, called Lab-on-a-chip, are proposed as a solution for improvement of cytotoxicity analysis of new compounds. Here, we present the evaluation of cytotoxic properties of graphene oxide (GO) on breast, liver and colon cancer cell line in a microfluidic system in two spatial models (2D and 3D). Before cell introduction, the microchambers surface was modified by the fibronectin (2D, monolayer) and poly(vinyl alcohol) (3D, spheroids) covering. After spheroid creation (3D) and cell attachment (2D, monolayer) the selected concentration of GO was introduced into microsystems. Then monolayer and spheroids viability/proliferation using alamarBlue® assay and standard microplate reader was checked for three days. Moreover, in every day of the culture, the morphological changes of cells were determined using microscopic analysis. Additionally, on the last day of the culture differential staining using Calcein AM and Propidium iodide were performed. We were able to note that the GO has an influence on all tested cell line viability in both monolayer and spheroid arrangement. We showed that GO caused higher viability/proliferation decrease for spheroids than a monolayer (this was observed for all tested cell lines). Higher cytotoxicity of GO on spheroid culture can be caused by different geometry of the microchambers for 2D and 3D cell cultures. Probably, GO was removed from the flat microchambers for 2D culture. Those results were also confirmed by differential staining. Comparing our results with the studies conducted in the macroscale, we also proved that the cytotoxic properties of GO are changed depending on the cell culture conditions (static/ flow).

Keywords: cytotoxicity, graphene oxide, monolayer, spheroid

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Authors: Ahmed Emad Ahmed, Mostafa El Abd, Ahmed Wakeb, Moahmmed Eissa

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Soil modification and reinforcing aims to increase soil shear strength and stiffness. In this report, different amounts of cement were added to the soil to explore its effect on shear strength and penetration using 3 tests. The first test is proctor compaction test which was conducted to determine the optimal moisture content and maximum dry density. The second test was direct shear test which was conducted to measure shear strength of soil. The third experiment was California bearing ratio test which was done to measure the penetration in soil. Each test was done different amount of times using different amounts of cement. The results from every test show that cement improve soil shear strength properties and stiffness.

Keywords: soil stabilized, soil, mechanical properties of soil, soil stabilized with a portland cement

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Authors: Seon Yeong Byeon, Hwa Sung Shin

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Nanofibrous sheets are of interest in the beauty industries due to the properties of moisturizing, adhesion to skin and delivery of nutrient materials. The benefit and function of the cosmetic products should not be considered without safety thus a non-toxic manufacturing process is ideal when fabricating the products. In this study, we have developed cosmetic patches consisting of alginate and Spirulina extract, a marine resource which has antibacterial and antioxidant effects, without addition of harmful cross-linkers. The patches obtained their structural stabilities by layer-upon-layer electrospinning of an alginate layer on a formerly spread polycaprolactone (PCL) layer instead of crosslinking method. The morphological characteristics, release of Spirulina extract, water absorption, skin adhesiveness and cytotoxicity of the double-layered patches were assessed. The image of scanning electron microscopy (SEM) showed that the addition of Spirulina extract has made the fiber diameter of alginate layers thinner. Impregnation of Spirulina extract increased their hydrophilicity, moisture absorption ability and skin adhesive ability. In addition, wetting the pre-dried patches resulted in releasing the Spirulina extract within 30 min. The patches were detected to have no cytotoxicity in the human keratinocyte cell-based MTT assay, but rather showed increased cell viability. All the results indicate the bioactive and hydro-adhesive double-layered patches have an excellent applicability to bioproducts for personal skin care in the trend of ‘A mask pack a day’.

Keywords: alginate, cosmetic patch, electrospun nanofiber, polycaprolactone, Spirulina extract

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Authors: Hediye Saglam

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This research has been carried out to bring in the 6 acquisitions in the 2014 Preschool Teaching Programme of the Turkish Ministry of Education through the method of analogy. This research is practiced based on the experimental pattern with pre-test and final test controlling groups. The working group of the study covers the group between 5-6 ages. The study takes 5 weeks including the 2 weeks spent for pre-test and the final test. It is conducted with the preschool teacher who gives the lesson along with the researcher in the in-class and out-of-class rainbow experiments of the students for 5 weeks. 'One Sample T Test' is used for the evaluation of the pre-test and final test. SPSS 17 programme is applied for the analysis of the data. Results: As an outcome of the study it is observed that analogy method affects children’s learning of the rainbow. For this very reason teachers should receive inservice training for different methods and techniques like analogy. This method should be included in preschool education programme and should be applied by teachers more often.

Keywords: acquisitions of preschool education programme, analogy method, pre-test/final test, rainbow experiments

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Authors: Amit Verma, Simranjeet Kaur, Sandeep Kaur

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Test Case Prioritization (TCP) has gained wide spread acceptance as it often results in good quality software free from defects. Due to the increase in rate of faults in software traditional techniques for prioritization results in increased cost and time. Main challenge in TCP is difficulty in manually validate the priorities of different test cases due to large size of test suites and no more emphasis are made to make the TCP process automate. The objective of this paper is to detect the priorities of different test cases using an artificial neural network which helps to predict the correct priorities with the help of back propagation algorithm. In our proposed work one such method is implemented in which priorities are assigned to different test cases based on their frequency. After assigning the priorities ANN predicts whether correct priority is assigned to every test case or not otherwise it generates the interrupt when wrong priority is assigned. In order to classify the different priority test cases classifiers are used. Proposed algorithm is very effective as it reduces the complexity with robust efficiency and makes the process automated to prioritize the test cases.

Keywords: test case prioritization, classification, artificial neural networks, TF-IDF

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Authors: Shunpu Zhang

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We propose a new multiple test called the minPOP test for testing multiple hypotheses simultaneously. Under the assumption that the test statistics are independent, we show that the minPOP test has higher global power than the existing multiple testing methods. We further propose a stepwise multiple-testing procedure based on the minPOP test and two of its modified versions (the Double Truncated and Left Truncated minPOP tests). We show that these multiple tests have strong control of the family-wise error rate (FWER). A method for finding the p-values of the proposed tests after adjusting for multiplicity is also developed. Simulation results show that the Double Truncated and Left Truncated minPOP tests, in general, have a higher number of rejections than the existing multiple testing procedures.

Keywords: multiple test, single-step procedure, stepwise procedure, p-value for multiple testing

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Authors: Nthatisi I. Molefe-Nyembe, Keisuke Suganuma, Oriel M. M. Thekisoe, Xuan Xuenan, Noboru Inoue

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African trypanosomosis is a devastating disease of animals caused by parasites of the genus Trypanosoma negatively affecting the economic status of more than 36 African countries. Few available drugs for the treatment of trypanosomosis remain inaccessible in remote areas, are associated with severe toxicity and most importantly, resistance has widely developed against their usage. Therefore, safe, effective and easily administrable drugs are urgently in need. The objective of the current study was to determine efficacy of azithromycin (AZM), on T. congolense, T. brucei brucei in vitro and in vivo. A 96 well luciferase assay was conducted to determine the trypanocidal effect of AZM on T. congolense, T. b. brucei and T. evansi as well as the cytotoxicity effect on the MDBK and NIH 3T3 cells. Additionally, TEM analysis was conducted to determine the morphological alteration on the AZM treated samples. Mice were infected with T. congolense and T. b. brucei and orally treated with AZM for 7 and 28 days referred to as the short and the long-term treatment. The in vitro IC50 values of AZM on T. congolense, T. b. brucei and T. evansi was 0.19 ± 0.17; 3.69 ± 2.26 and 1.81 ± 1.82 μg/mL, respectively, while the cytotoxicity effects values were greater than 25 μg/mL. A vacuole-like structure was observed in the TEM imaging of AZM treated T. congolense, while the presence of glycosomes and acidocalcisome-like structured were detected in T. b. brucei samples. In vivo, AZM was more effective against T. congolense infected mice than T. b. brucei. In conclusion, AZM exhibited the trypanocidal effects on T. congolense and T. b. brucei infected mice. However, further studies are necessary to determine the metabolic pathway responsible for the observed efficacy.

Keywords: animal trypanosomosis, azithromycin, oral administration, trypanosoma congolense

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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: Chen Li-Ching

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The multinomial logistic regression model is used popularly for inferring the relationship of risk factors and disease with multiple categories. This study based on the discrepancy between the nonparametric maximum likelihood estimator and semiparametric maximum likelihood estimator of the cumulative distribution function to propose a Kolmogorov-Smirnov type test statistic to assess adequacy of the multinomial logistic regression model for case-control data. A bootstrap procedure is presented to calculate the critical value of the proposed test statistic. Empirical type I error rates and powers of the test are performed by simulation studies. Some examples will be illustrated the implementation of the test.

Keywords: case-control studies, goodness-of-fit test, Kolmogorov-Smirnov test, multinomial logistic regression

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Authors: Andualem Workie

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In this study, we examined the sinterability and bioactivity of MgO-SiO₂-P₂O₅-CaO-CaF₂ glass compositions created through spray pyrolysis. We evaluated the bioactivity of the materials by immersing them for varying periods of time in simulated bodily fluid (SBF) and found that bioactivity was related to the sintering temperature and soaking time. The material's pH value during immersion in SBF was within the range of 7.4-8.2, which is below 8.5 and improves compatibility and reduces toxicity in biological applications. We used X-ray diffraction and scanning electron microscopy to determine the phase compositions and morphologies of the samples and found that the 1100°C sintered A-W GC sample exhibited the highest bioactivity after soaking in SBF. This sample was dominated by fluorapatite, wollastonite, and whitlockite crystals scattered throughout the glass matrix. The crystallinity (%) of the A-W GC increased as its bioactivity improved, making it more suitable for use in pharmaceutical applications. We also conducted a cytotoxicity test on A-W GC samples sintered at different temperatures and found that the glass-ceramics were non-toxic to MC3T3-E1 cells at all extraction concentrations, except for those sintered at 700°C at concentrations of 250, 200, and 150 mg/ml where cell viability (%) was below the threshold of 70%.

Keywords: apatite wollastonite glass ceramics, bioactivity, calcination, cell viability

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Authors: Lesly Y Carmona-Sarabia, Luisa Barraza-Vergara, Vilmalí López-Mejías, Wandaliz Torres-García, Maribella Domenech-Garcia, Madeline Torres-Lugo

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Biosensors are used in many applications providing real-time monitoring to treat long-term conditions. Thus, understanding the physicochemical properties and biological side effects on the skin of polymers (e. g., poly(methyl methacrylate), PMMA) employed in the fabrication of wearable biosensors is crucial for the selection of manufacturing materials within this field. The PMMA (hydrophobic and thermoplastic polymer) is commonly employed as a coating material or substrate in the fabrication of wearable devices. The cytotoxicityof PMMA (including residual monomers or degradation products) on the skin, in terms of cells and tissue, is required to prevent possible adverse effects (cell death, skin reactions, sensitization) on human health. Within this work, accelerated aging of PMMA (Mw ~ 15000) through thermal and photochemical degradation was under-taken. The accelerated aging of PMMA was carried out by thermal (200°C, 1h) and photochemical degradation (UV-Vis, 8-15d) adapted employing ISO protocols (ISO-10993-12, ISO-4892-1:2016, ISO-877-1:2009, ISO-188: 2011). In addition, in vitro cytotoxicity evaluation of PMMA degradation products was performed using NIH3T3 fibroblast cells to assess the response of skin tissues (in terms of cell viability) exposed with polymers utilized to manufacture wearable biosensors, such as PMMA. The PMMA (Mw ~ 15000) before and after accelerated aging experiments was characterized by thermal gravimetric analysis (TGA), differential scanning calorimetric (DSC), powder X-ray diffractogram (PXRD), and scanning electron microscopy-energy dispersive spectroscopy (SEM-EDS) to determine and verify the successful degradation of this polymer under the specific conditions previously mention. The degradation products were characterized through nuclear magnetic resonance (NMR) to identify possible byproducts generated after the accelerated aging. Results demonstrated a percentage (%) weight loss between 1.5-2.2% (TGA thermographs) for PMMA after accelerated aging. The EDS elemental analysis reveals a 1.32 wt.% loss of carbon for PMMA after thermal degradation. These results might be associated with the amount (%) of PMMA degrade after the accelerated aging experiments. Furthermore, from the thermal degradation products was detected the presence of the monomer and methyl formate (low concentrations) and a low molecular weight radical (·COOCH3) in higher concentrations by NMR. In the photodegradation products, methyl formate was detected in higher concentrations. These results agree with the proposed thermal or photochemical degradation mechanisms found in the literature.1,2 Finally, significant cytotoxicity on the NIH3T3 cells was obtained for the thermal and photochemical degradation products. A decrease in cell viability by > 90% (stock solutions) was observed. It is proposed that the presence of byproducts (e.g. methyl formate or radicals such as ·COOCH₃) from the PMMA degradation might be responsible for the cytotoxicity observed in the NIH3T3 fibroblast cells. Additionally, experiments using skin models will be employed to compare with the NIH3T3 fibroblast cells model.

Keywords: biosensors, polymer, skin irritation, degradation products, cell viability

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Authors: Kritika Kulhari, Aparna Sahu

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Test anxiety results from concerns about failure in examinations or evaluative situations. Attentional biases are known to pronounce the symptomatic expression of test anxiety. In recent times, the inattentional blindness (IB) paradigm has shown promise as an attention bias modification treatment (ABMT) for anxiety by overcoming practice and expectancy effects which preexisting paradigms fail to counter. The IB paradigm assesses the inability of an individual to attend to a stimulus that appears suddenly while indulging in a perceptual discrimination task. The present study incorporated an IB task with three critical items (book, face, and triangle) appearing randomly in the perceptual discrimination task. Attentional biases were assessed as detection and identification of the critical item. The sample (N = 50) consisted of low test anxiety (LTA) and high test anxiety (HTA) groups based on the reactions to tests scale scores. Test threat manipulation was done with pre- and post-test assessment of test anxiety using the State Test Anxiety Inventory. A mixed factorial design with gender, test anxiety, presence or absence of test threat, and critical items was conducted to assess their effects on attentional biases. Results showed only a significant main effect for test anxiety on detection with higher accuracy of detection of the critical item for the LTA group. The study presents promising results in the realm of ABMT for test anxiety.

Keywords: attentional bias, attentional bias modification treatment, inattentional blindness, test anxiety

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Authors: T. Scattolin, E. Cavarzerani, F. Visentin, F. Rizzolio

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Organopalladium compounds have recently attracted attention for their high stability even under physiological conditions and, above all, for their remarkable in vitro cytotoxicity towards cisplatin-resistant cell lines. Among the organopalladium derivatives, those bearing at least one N-heterocyclic carbene ligand (NHC) and the Pd(II)-η³-allyl fragment have exhibited IC₅₀ values in the micro and sub-micromolar range towards several cancer cell lines in vitro and in some cases selectivity towards cancerous vs. non-tumorigenic cells. Herein, a selection of allyl and indenyl palladates were synthesized using a solvent-free method consisting of grinding the corresponding palladium precursors with different saturated and unsaturated azolium salts. All compounds have been fully characterized by NMR, XRD and elemental analyses. The intramolecular H, Cl interaction has been elucidated and quantified using the Voronoi Deformation Density scheme. Most of the complexes showed excellent cytotoxicity towards ovarian cancer cell lines, with I₅₀ values comparable to or even lower than cisplatin. Interestingly, the potent anticancer activity was also confirmed in a high-serous ovarian cancer (HGSOC) patient-derived tumoroid, with a clear superiority of this class of compounds over classical platinum-based agents. Finally, preliminary enzyme inhibition studies of the synthesized palladate complexes against the model TrxR show that the compounds have high activity comparable to or even higher than auranofin and classical Au(I) NHC complexes. Based on such promising data, further in vitro and in vivo experiments and in-depth mechanistic studies are ongoing in our laboratories.

Keywords: anticancer activity, palladium complexes, organoids, indenyl and allyl ligands

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Authors: Gyati Shilakari Asthana, Abhay Asthana

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Purpose: The therapeutic potential of oligonucleotide (ODN) is primarily dependent upon its safe and efficient delivery to specific cells overcoming degradation and maximizing cellular uptake in vivo. The present study is focused to design low molecular weight chitosan nanoconstructs to meet the requirements of safe and effectual delivery of ODNs. LMW-chitosan is a biodegradable, water soluble, biocompatible polymer and is useful as a non-viral vector for gene delivery due to its better stability in water. Methods: LMW chitosan ODN nanoparticles (CHODN NPs) were formulated by self assembled method using various N/P ratios (moles ratio of amine groups of CH to phosphate moieties of ODNs; 0.5:1, 1:1, 3:1, 5:1 and 7:1) of CH to ODN. The developed CHODN NPs were evaluated with respect to gel retardation assay, particle size, zeta potential and cytotoxicity and transfection efficiency. Results: Complete complexation of CH/ODN was achieved at the charge ratio of 0.5:1 or above and CHODN NPs displayed resistance against DNase I. On increasing the N/P ratio of CH/ODN, particle size of the NPs decreased whereas zeta potential (ZV) value increased. No significant toxicity was observed at all CH concentrations. The transfection efficiency was increased on increasing N/P ratio from 1:1 to 3:1, whereas it was decreased with further increment in N/P ratio upto 7:1. Maximum transfection of CHODN NPs with both the cell lines (Raw 267.4 cells and Hela cells) was achieved at N/P ratio of 3:1. The results suggest that transfection efficiency of CHODN NPs is dependent on N/P ratio. Conclusion: Thus the present study states that LMW chitosan nanoparticulate carriers would be acceptable choice to improve transfection efficiency in vitro as well as in vivo delivery of oligonucleotide.

Keywords: LMW-chitosan, chitosan nanoparticles, biocompatibility, cytotoxicity study, transfection efficiency, oligonucleotide

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Authors: Saule Mussabekova, Samal Abzhanova

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The Kazakh Language Fluency Test, based on the IELTS exam, was implemented in 2012 at Nazarbayev University in Astana, Kazakhstan. We would like to share our experience in developing this exam and some exam results with other language instructors. In this paper, we will cover all these peculiarities and their related issues. The Kazakh Language Fluency Test is a young exam. During its development, we faced many difficulties. One of the goals of the university and the country is to encourage fluency in the Kazakh language for all citizens of the Republic. Nazarbayev University has introduced a Kazakh language program to assist in achieving this goal. This policy is one-step in ensuring that NU students have a thorough understanding of the Kazakh language through a fluency test based on the International English Language Testing System (IELTS). The Kazakh Language Fluency Test exam aims to determine student’s knowledge of Kazakh language. The fact is that there are three types of students at Nazarbayev University: Kazakh-speaking heritage learners, Russian-speaking and English-speaking students. Unfortunately, we have Kazakh students who do not speak Kazakh. All students who finished school with Russian language instruction are given Kazakh Language Fluency Test in order to determine their Kazakh level. After the test exam, all students can choose appropriate Kazakh course: Basic Kazakh, Intermediate Kazakh and Upper-Intermediate Kazakh. The Kazakh Language Fluency Test consists of four parts: Listening, Reading, Writing and Speaking. They are taken on the same day in the abovementioned order.

Keywords: diagnostic test, kazakh language, placement test, test result

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Authors: Jin Koo

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The main purpose of this study is to investigate the comparability of student scores obtained from Test Center (TC) vs. Online-Proctored (OP) Delivery in the three subject areas of Verbal, Reading, and Mathematics for each level (Middle and Upper). Also, this study examines whether there is an interaction effect between test deliveries (TC vs. OP) and gender/ethnicity/ability level in each subject area. The test used in this study is a multiple-choice standardized test for students in grades 5-11. For this study, data were collected during the 2022-23 test administration. This research used a one-factor between-subjects ANOVA and Cohen’s d to compare the TC and OP groups’ test means for each level and each subject area. Also, 2-factor between-subjects ANOVAs were conducted to investigate examinee characteristics: gender (male and female), ethnicity (African-American, Asian, Hispanic, Multi-racial, and White), and ability level (low, average, and high-ability groups). The author found that students’ test scores in some subject areas varied between TC and OP test deliveries by gender, ethnicity, and ability level, meaning that gender, ethnicity, and ability level were related to the score difference. These results will be discussed according to the current testing systems.

Keywords: ability level, ethnicity, gender, online-proctored delivery, standardized test, test center

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Authors: Prantasi Harmi Tjahjanti, Iswanto Iswanto, Edi Widodo, Sholeh Pamuji

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Materials of the thermoplastic polymer when they break is usually thrown away, or is recycled which requires a long process. The purpose of this study is to splice the broken thermoplastic polymer using hot gas welding with different variations of welding wire/electrodes. Materials of thermoplastic polymer used are Polyethylene (PE), Polypropylene (PP), and Polyvinyl chloride (PVC) by using welding wire like the three materials. The method is carried out by using hot gas welding; there are two materials that cannot be connected, namely PE with PVC welding wire, and PP with PVC welding wire. The permeable liquid penetrant test is PP with PE welding wire, and PVC with PE welding wire. The best bending test result with the longest elongation is PE with PE welding wire with a bending test value of 179.03 kgf/mm². The microstructure was all described in Scanning Electron Microscopy (SEM) observations.

Keywords: thermoplastic polymers, bending test, polyethylene (PE), polypropylene (PP), polyvinyl chloride (PVC), hot gas welding, bending test

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Authors: Nancy M. El-Baz, Laila Ziko, Rania Siam, Wael Mamdouh

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Cancer is one of the most devastating diseases, and has over than 10 million new cases annually worldwide. Despite the effectiveness of chemotherapeutic agents, their systemic toxicity and non-selective anticancer actions represent the main obstacles facing cancer curability. Due to the effective enhanced permeability and retention (EPR) effect of nanomaterials, nanoparticles (NPs) have been used as drug nanocarriers providing targeted cancer drug delivery systems. In addition, several inorganic nanoparticles such as silver (AgNPs) nanoparticles demonstrated a potent anticancer activity against different cancers. The present study aimed at formulating core-shell inorganic NPs-based combinatorial therapy based on combining the anticancer activity of AgNPs along with doxorubicin (DOX) and evaluating their cytotoxicity on MCF-7 breast cancer cells. These inorganic NPs-based combinatorial therapies were designed to (i) Target and kill cancer cells with high selectivity, (ii) Have an improved efficacy/toxicity balance, and (iii) Have an enhanced therapeutic index when compared to the original non-modified DOX with much lower dosage The in-vitro cytotoxicity studies demonstrated that the NPs-based combinatorial therapy achieved the same efficacy of non-modified DOX on breast cancer cell line, but with 96% reduced dose. Such reduction in DOX dose revealed that the combination between DOX and NPs possess a synergic anticancer activity against breast cancer. We believe that this is the first report on a synergic anticancer effect at very low dose of DOX against MCF-7 cells. Future studies on NPs-based combinatorial therapy may aid in formulating novel and significantly more effective cancer therapeutics.

Keywords: nanoparticles-based combinatorial therapy, silver nanoparticles, doxorubicin, breast cancer

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Authors: Mikako Nobuhara

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The Test of English for International Communication (TOEIC) test, conducted by the Institute for International Business Communication (IIBC), has a huge impact on education in Japan. Almost all college students have to submit their TOEIC test scores when applying for entry-level jobs at companies. In addition, an increasing number of colleges are encouraging students to have a global vision. For this specific reason, studying for the TOEIC test is essential for English as a second language (ESL) learner to develop English communication skills. This study shows that studying by using some workbooks about the listening section of the TOEIC test clearly helps ESL learners to develop their listening skills. For this purpose, the listening test scores before and after classroom sessions were analyzed for each student. Students obtained higher scores in the listening section of the test and improved their English listening skills at the end of all the classroom sessions. In conclusion, it is important for English teachers to achieve the following objectives: (1) facilitate the learning of effective methods for correctly solving questions based on listening skills and (2) prepare listening tasks for reading aloud so as to keep up with the original speed, which is required for solving questions in the TOEIC test.

Keywords: education, ESL, listening skills, TOEIC test

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Authors: Yoshinori Wakatake

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Many defects after the release of software packages are caused due to omissions of sufficient test items in test specifications. Poor test specifications are detected by manual review, which imposes a high human load. The prevention of omissions depends on the end-user awareness of test specification writers. If test specifications were written while envisioning the behavior of end-users, the number of omissions in test items would be greatly reduced. The paper pays attention to the point that writers who can achieve it differ from those who cannot in not only the description richness but also their gaze information. It proposes a method to estimate the degree of user-awareness of writers through the analysis of their gaze information when writing test specifications. We conduct an experiment to obtain the gaze information of a writer of the test specifications. Test specifications are automatically classified using gaze information. In this method, a Random Forest model is constructed for the classification. The classification is highly accurate. By looking at the explanatory variables which turn out to be important variables, we know behavioral features to distinguish test specifications of high quality from others. It is confirmed they are pupil diameter size and the number and the duration of blinks. The paper also investigates test specifications automatically classified with gaze information to discuss features in their writing ways in each quality level. The proposed method enables us to automatically classify test specifications. It also prevents test item omissions, because it reveals writing features that test specifications of high quality should satisfy.

Keywords: blink, eye tracking, gaze information, pupil diameter, quality improvement, specification document, user-awareness

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Authors: Renata Rank Miranda, Arandi Ginane Bezerra, Ciro Alberto Oliveira Ribeiro, Marco AntôNio Ferreira Randi, Carmen Lúcia Voigt, Lilian Skytte, Kaare Lund Rasmussen, Francisco Filipak Neto, Frank Kjeldsen

Abstract:

Synergetic and antagonistic effects of drugs are well-known concerns in pharmacological assessments of dose and toxicity. Similar approach should be used in assessing cellular uptake and cytotoxicity of nanoparticles. Since nanoparticles are released into the aquatic environment they may interact with existing xenobiotics. Here we used biochemical assays and quantitative proteomics to assess the cytotoxicity of silver nanoparticles (AgNP) when human hepatoma HepG2 cells were co-exposed to 2 nm AgNP together with either Cd2+ or Hg2+ ions. Time-course experiments (2h, 4h, and 24h) were conducted to assess the first response to the exposure studies. The general trend was that a synergetic toxicological response was observed in cells exposed to both AgNP and Cd2+ or Hg2+, with AgNP and Cd2+ being more toxic. This was observed by a significant increase in the ROS and superoxide level of >35% in the case of AgNP+Cd2+ compared to the sum of responses of AgNP and Cd2+, individually. Metabolic activity and viability also dropped more for AgNP+Cd2+ (>10%) than for AgNP and Cd2+ combined. We used inductively coupled plasma mass spectrometry to investigate if AgNP facilitates larger influx of toxic metal ions into HepG2 cells. Only Hg2+ ions was found to be more efficiently engulfed as the concentration of Hg2+ was found 2.8 times larger compared to exposure experiments with only Hg2+. This effect was not observed for Cd2+. We now continue with deep proteomics studies to obtain wider details on the mechanism of the toxicity related to AgNP, Cd2+, and AgNP+Cd2+, respectively.

Keywords: nanotoxicology, silver nanoparticles, proteomics, human cell line

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Authors: Gyati Shilakari Asthana, Abhay Asthana, Dharm Veer Kohli, Suresh Prasad Vyas

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Purpose: The therapeutic potential of oligonucleotide (ODN) is primarily dependent upon its safe and efficient delivery to specific cells overcoming degradation and maximizing cellular uptake in vivo. The present study is focused to design low molecular weight chitosan nanoconstructs to meet the requirements of safe and effectual delivery of ODNs. LMW-chitosan is a biodegradable, water soluble, biocompatible polymer and is useful as a non-viral vector for gene delivery due to its better stability in water. Methods: LMW chitosan ODN nanoparticles (CHODN NPs) were formulated by self-assembled method using various N/P ratios (moles ratio of amine groups of CH to phosphate moieties of ODNs; 0.5:1, 1:1, 3:1, 5:1, and 7:1) of CH to ODN. The developed CHODN NPs were evaluated with respect to gel retardation assay, particle size, zeta potential and cytotoxicity and transfection efficiency. Results: Complete complexation of CH/ODN was achieved at the charge ratio of 0.5:1 or above and CHODN NPs displayed resistance against DNase I. On increasing the N/P ratio of CH/ODN, the particle size of the NPs decreased whereas zeta potential (ZV) value increased. No significant toxicity was observed at all CH concentrations. The transfection efficiency was increased on increasing N/P ratio from 1:1 to 3:1, whereas it was decreased with further increment in N/P ratio upto 7:1. Maximum transfection of CHODN NPs with both the cell lines (Raw 267.4 cells and Hela cells) was achieved at N/P ratio of 3:1. The results suggest that transfection efficiency of CHODN NPs is dependent on N/P ratio. Conclusion: Thus the present study states that LMW chitosan nanoparticulate carriers would be acceptable choice to improve transfection efficiency in vitro as well as in vivo delivery of oligonucleotide.

Keywords: LMW-chitosan, chitosan nanoparticles, biocompatibility, cytotoxicity study, transfection efficiency, oligonucleotide

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Authors: Emine Feyza Şükür, Mevlüt Türköz, Murat Dilmeç, Hüseyin Selçuk Halkacı

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In this study, a warm bulge test system was designed, built and experimentally validated to perform warm bulge tests with all necessary systems. In addition, performance of each sub-system is validated through repeated production and/or test runs as well as through part quality measurements. Validation and performance tests were performed to characterize the repeatability of the system. As a result of these tests, the desired temperature distribution on the sheet metal was obtained by the heating systems and the good repeatability of the bulge tests was obtained. Consequently, this study is expected to provide other researchers and manufacturer with a set of design and process guidelines to develop similar systems.

Keywords: design, test unit, warm bulge test unit, validation test

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Authors: Arslan Ellahi, Syed Amjad Hussain

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Software Testing is a vital process in software development life cycle. We can attain the quality of software after passing it through software testing phase. We have tried to find out automatic test data generation techniques that are a key research area of software testing to achieve test automation that can eventually decrease testing time. In this paper, we review some of the approaches presented in the literature which use evolutionary search based algorithms like Genetic Algorithm, Particle Swarm Optimization (PSO), etc. to validate the test data generation process. We also look into the quality of test data generation which increases or decreases the efficiency of testing. We have proposed test data generation techniques for model-based testing. We have worked on tuning and fitness function of PSO algorithm.

Keywords: search based, evolutionary algorithm, particle swarm optimization, genetic algorithm, test data generation

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Authors: Šárka Hradilová, Aleš Panáček, Radek Zbořil

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Nanotechnologies are considered the most promising fields with high added value, brings new possibilities in various sectors from industry to medicine. With the growing of interest in nanomaterials and their applications, increasing nanoparticle production leads to increased exposure of people and environment with ‘human made’ nanoparticles. Nanoparticles (NPs) are clusters of atoms in the size range of 1–100 nm. Metal nanoparticles represent one of the most important and frequently used types of NPs due to their unique physical, chemical and biological properties, which significantly differ from those of bulk material. Biological properties including toxicity of metal nanoparticles are generally determined by their size, size distribution, shape, surface area, surface charge, surface chemistry, stability in the environment and ability to release metal ions. Therefore, the biological behavior of NPs and their possible adverse effect cannot be derived from the bulk form of material because nanoparticles show unique properties and interactions with biological systems just due to their nanodimensions. Silver and gold NPs are intensively studied and used. Both can be used for instance in surface enhanced Raman spectroscopy, a considerable number of applications of silver NPs is associated with antibacterial effects, while gold NPs are associated with cancer treatment and bio imaging. Antibacterial effects of silver ions are known for centuries. Silver ions and silver-based compounds are highly toxic to microorganisms. Toxic properties of silver NPs are intensively studied, but the mechanism of cytoxicity is not fully understood. While silver NPs are considered toxic, gold NPs are referred to as toxic but also innocuous for eukaryotic cells. Therefore, gold NPs are used in various biological applications without a risk of cell damaging, even when we want to suppress the growth of cancer cells. Thus, gold NPs are toxic or harmless. Because most studies comparing particles of various sizes prepared in various ways, and testing is performed on different cell lines, it is very difficult to generalize. The novelty and significance of our research is focused to the complex biological effects of silver and gold NPs prepared by the same method, have the same parameters and the same stabilizer. That is why we can compare the biological effects of pure nanometals themselves based on their chemical nature without the influence of other variable. Aim of our study therefore is to compare the cytotoxic effect of two types of noble metal NPs focusing on the mechanisms that contribute to cytotoxicity. The study was conducted on murine fibroblasts by selected common used tests. Each of these tests monitors the selected area related to toxicity and together provides a comprehensive view on the issue of interactions of nanoparticles and living cells.

Keywords: cytotoxicity, gold nanoparticles, mechanism of cytotoxicity, silver nanoparticles

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