Search results for: in vitro release

Authors: Anna Jesionek, Aleksandra Szreniawa-Sztajnert, Zbigniew Jaremicz, Adam Kokotkiewicz, Natalia Filipowicz, Renata Ochocka, Bozena Zabiegala, Maria Luczkiewicz

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Rhododendron tomentosum (formerly Ledum palustre), an evergreen shrub grows in peaty soils in northern Europe, Asia and North America. In Poland, it is classified as an endangered species not only due to the drainage of wetlands, but also to the excessive collection of this repellent plant by human. The other valuable biological properties of R. tomentosum, used for years in folk medicine, include anti-inflammatory, analgesic and anti-microbial activity, conditioned by the essential oil content. Taking into account the importance of biodiversity and the potential therapeutic application, it was decided to establish, for the first time, the micropropagation protocol for R. tomentosum, for ex-situ conservation of this endangered species as well as to obtain the continuous source of in vivo and in-vitro plant material for further studies. This object was achieved by the selection of the explant and the media, which were modified within the scope of mineral composition, sugar content, pH and the growth regulators. As a result, the four-stage micropropagation protocol for R. tomentosum was specified, including shoot multiplication, elongation, rooting and ex-vitro adaptation. The genetic identification of the examined species and the compatibility of progeny plants with maternal ones was tested with molecular biology methods. Moreover, during the research process, the chemical composition of initial and regenerated plant and in vitro shoots was controlled in terms of volatile fraction by phytochemical analysis (GC and TLC methods). The correctness of the micropropagation procedure was confirmed by both types of studies.

Keywords: ex situ conservation, Ledum palustre, micropropagation, Rhododendron tomentosum

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Authors: Sijing Xiong, Ran Su, Lit-Hsin Loo, Daniele Zink

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The kidney is a major target for toxic effects of drugs, industrial and environmental chemicals and other compounds. Typically, nephrotoxicity is detected late during drug development, and regulatory animal models could not solve this problem. Validated or accepted in silico or in vitro methods for the prediction of nephrotoxicity are not available. We have established the first and currently only pre-validated in vitro models for the accurate prediction of nephrotoxicity in humans and the first predictive platforms based on renal cells derived from human pluripotent stem cells. In order to further improve the efficiency of our predictive models, we recently developed a high content screening (HCS) platform. This platform employed automated imaging in combination with automated quantitative phenotypic profiling and machine learning methods. 129 image-based phenotypic features were analyzed with respect to their predictive performance in combination with 44 compounds with different chemical structures that included drugs, environmental and industrial chemicals and herbal and fungal compounds. The nephrotoxicity of these compounds in humans is well characterized. A combination of chromatin and cytoskeletal features resulted in high predictivity with respect to nephrotoxicity in humans. Test balanced accuracies of 82% or 89% were obtained with human primary or immortalized renal proximal tubular cells, respectively. Furthermore, our results revealed that a DNA damage response is commonly induced by different PTC-toxicants with diverse chemical structures and injury mechanisms. Together, the results show that the automated HCS platform allows efficient and accurate nephrotoxicity prediction for compounds with diverse chemical structures.

Keywords: high content screening, in vitro models, nephrotoxicity, toxicity prediction

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Authors: Biswabandhu Bankura, Srikanta Guria, Madhusudan Das

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Purpose: Thyroid peroxidase (TPO) is the key enzyme in the biosynthesis of thyroid hormones. We aimed to identify the spectrum of mutations in the TPO gene leading to hypothyroidism in the population of West Bengal to establish the genetic etiology of the disease. Methods: 200 hypothyroid patients (case) and their corresponding sex and age matched 200 normal individuals (control) were screened depending on their clinical manifestations. Genomic DNA was isolated from peripheral blood samples and TPO gene (Exon 7 to Exon 14) was amplified by PCR. The PCR products were subjected to sequencing to identify mutations. Results: Single nucleotide changes such as Glu 641 Lys, Asp 668 Asn, Thr 725 Pro, Asp 620 Asn, Ser 398 Thr, and Ala 373 Ser were found. Changes in the TPO were assayed in vitro to compare mutant and wild-type activities. Five mutants were enzymatically inactive in the guaiacol and iodide assays. This is a strong indication that the mutations are present at crucial positions of the TPO gene, resulting in inactivated TPO. Key Findings: The results of this study may help to develop a genetic screening protocol for goiter and hypothyroidism in the population of West Bengal.

Keywords: thyroid peroxidase, hypothyroidism, mutation, in vitro assay, transfection

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Authors: Biswabandhu Bankura, Srikanta Guria, Madhusudan Das

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Purpose: Thyroid peroxidase (TPO) is the key enzyme in the biosynthesis of thyroid hormones. We aimed to identify the spectrum of mutations in the TPO gene leading to hypothyroidism in the population of West Bengal to establish the genetic etiology of the disease. Methods: 200 hypothyroid patients (case) and their corresponding sex and age matched 200 normal individuals (control) were screened depending on their clinical manifestations. Genomic DNA was isolated from peripheral blood samples and TPO gene (Exon 7 to Exon 14) was amplified by PCR. The PCR products were subjected to sequencing to identify mutations. Results: Single nucleotide changes such as Glu 641 Lys, Asp 668 Asn, Thr 725 Pro, Asp 620 Asn, Ser 398 Thr, and Ala 373 Ser were found. Changes in the TPO were assayed in vitro to compare mutant and wild-type activities. Five mutants were enzymatically inactive in the guaiacol and iodide assays. This is a strong indication that the mutations are present at crucial positions of the TPO gene, resulting in inactivated TPO. Key Findings: The results of this study may help to develop a genetic screening protocol for goiter and hypothyroidism in the population of West Bengal.

Keywords: thyroid peroxidase, hypothyroidism, mutation, in vitro assay, transfection

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Authors: Pornthep Rachnavy

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The purpose of this paper is twofold. First, we believe that there are a significant relationship between sculling angle and sculling style among adaptive rowing. Second, we introduce a methodology used for adaptive rowing, namely simulation, to identify effectiveness of adaptive rowing. For our study we simulate the arms only single scull of adaptive rowing. The method for rowing fastest under the 1000 meter was investigated by study sculling angle using the simulation modeling. A simulation model of a rowing system was developed using the Matlab software package base on equations of motion consist of many variation for moving the boat such as oars length, blade velocity and sculling style. The boat speed, power and energy consumption on the system were compute. This simulation modeling can predict the force acting on the boat. The optimum sculling angle was performing by computer simulation for compute the solution. Input to the model are sculling style of each rower and sculling angle. Outputs of the model are boat velocity at 1000 meter. The present study suggests that the optimum sculling angle exist depends on sculling styles. The optimum angle for blade entry and release with respect to the perpendicular through the pin of the first style is -57.00 and 22.0 degree. The optimum angle for blade entry and release with respect to the perpendicular through the pin of the second style is -57.00 and 22.0 degree. The optimum angle for blade entry and release with respect to the perpendicular through the pin of the third style is -51.57 and 28.65 degree. The optimum angle for blade entry and release with respect to the perpendicular through the pin of the fourth style is -45.84 and 34.38 degree. A theoretical simulation for rowing has been developed and presented. The results suggest that it may be advantageous for the rowers to select the sculling angles proper to sculling styles. The optimum sculling angles of the rower depends on the sculling styles made by each rower. The investigated of this paper can be concludes in three directions: 1;. There is the optimum sculling angle in arms only single scull of adaptive rowing. 2. The optimum sculling angles depend on the sculling styles. 3. Computer simulation of rowing can identify opportunities for improving rowing performance by utilizing the kinematic description of rowing. The freedom to explore alternatives in speed, thrust and timing with the computer simulation will provide the coach with a tool for systematic assessments of rowing technique In addition, the ability to use the computer to examine the very complex movements during rowing will help both the rower and the coach to conceptualize the components of movements that may have been previously unclear or even undefined.

Keywords: simulation, sculling, adaptive, rowing

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Authors: Steven Arcidiacono, Robert Stote, Erin Anderson, Molly Richards

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There are numerous standard test methods for antimicrobial textiles that measure activity against specific microorganisms. However, many times these results do not translate to the performance of treated textiles when worn by individuals. Standard test methods apply a single target organism grown under optimal conditions to a textile, then recover the organism to quantitate and determine activity; this does not reflect the actual performance environment that consists of polymicrobial communities in less than optimal conditions or interaction of the textile with the skin substrate. Here we propose the development of in vitro skin model method to bridge the gap between lab testing and wear studies. The model will consist of a defined polymicrobial community of 5-7 commensal microbes simulating the skin microbiome, seeded onto a solid tissue platform to represent the skin. The protocol would entail adding a non-commensal test organism of interest to the defined community and applying a textile sample to the solid substrate. Following incubation, the textile would be removed and the organisms recovered, which would then be quantitated to determine antimicrobial activity. Important parameters to consider include identification and assembly of the defined polymicrobial community, growth conditions to allow the establishment of a stable community, and choice of skin surrogate. This model could answer the following questions: 1) is the treated textile effective against the target organism? 2) How is the defined community affected? And 3) does the textile cause unwanted effects toward the skin simulant? The proposed model would determine activity under conditions comparable to the intended application and provide expanded knowledge relative to current test methods.

Keywords: antimicrobial textiles, defined polymicrobial community, in vitro skin model, skin microbiome

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Authors: Anbang Liu, Huaqing Xie, Zihua Wu, Xiaoxiao Yu, Yuanyuan Wang

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Due to the temperature-dependence and mutual coupling of thermoelectric parameters, the Thomson effect always exists, which is derived from temperature gradients during thermoelectric conversion. The synergistic effect between the Thomson effect and non-equilibrium heat transport of charge carriers leads to local heat absorption or release in thermoelements, thereby affecting its power generation performance and conversion efficiency. This study verified and analyzed the influence and mechanism of the Thomson effect on N-type skutterudite thermoelement through quasi-steady state testing under approximate vacuum conditions. The results indicate the temperature rise/fall of N-type thermoelement at any position is affected by Thomson heat release/absorption. Correspondingly, the Thomson effect also contributes advantageously/disadvantageously to the output power of N-type skutterudite thermoelement when the Thomson coefficients are positive/negative. In this work, the output power can be promoted or decreased maximally by more than 27% due to the presence of Thomson heat when the absolute value of the Thomson coefficient is around 36 μV/℃.

Keywords: Thomson effect, heat transport, thermoelectric conversion, numerical simulation

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Authors: Yang Fu, Jaime Rodrigo Navarro, Jose F. Monserrat, Faiza Bouchmal, Oscar Carrasco Quilis

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Cellular Vehicle to Everything (C-V2X) is considered a promising solution for future autonomous driving. From Release 16 to Release 17, the Third Generation Partnership Project (3GPP) has introduced the definitions and services for 5G New Radio (NR) V2X. Experience from previous generations has shown that establishing a simulator for C-V2X communications is an essential preliminary step to achieve reliable and stable communication links. This paper proposes a complete framework of a link-level simulator based on the 3GPP specifications for the Physical Sidelink Share Channel (PSSCH) of the 5G NR Physical Layer (PHY). In this framework, several algorithms in the receiver part, i.e., sliding window in channel estimation and Minimum Mean Square Error (MMSE)-based equalization, are developed. Finally, the performance of the developed PSSCH receiver is validated through extensive simulations under different assumptions.

Keywords: C-V2X, channel estimation, link-level simulator, sidelink, 3GPP

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Authors: Erwin San Juan Martinez, Miguel Angel Aguilar Mendez, Manuel Sandoval Villa, Libia Iris Trejo Tellez

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Some nanomaterials may improve the vegetal growth in certain concentration intervals, and could be used as nanofertilizers in order to increase crops yield, and decreasing the environmental pollution due to non-controlled use of conventional fertilizers, therefore the present investigation’s objective was to synthetize and characterize gelatin nanoparticles loaded with calcium generated through pulverization technique and be used as nanofertilizers. To obtain these materials, a fractional factorial design 27-4 was used in order to evaluate the largest number of factors (concentration of Ca2+, temperature and agitation time of the solution and calcium concentration, drying temperature, and % spray) with a possible effect on the size, distribution and morphology of nanoparticles. For the formation of nanoparticles, a Nano Spray-Dryer B - 90® (Buchi, Flawil, Switzerland), equipped with a spray cap of 4 µm was used. Size and morphology of the obtained nanoparticles were evaluated using a scanning electron microscope (JOEL JSM-6390LV model; Tokyo, Japan) equipped with an energy dispersive x-ray X (EDS) detector. The total quantification of Ca2+ as well as its release by the nanoparticles was carried out in an equipment of induction atomic emission spectroscopy coupled plasma (ICP-ES 725, Agilent, Mulgrave, Australia). Of the seven factors evaluated, only the concentration of fertilizer, % spray and concentration of polymer presented a statistically significant effect on particle size. Micrographs of SEM from six of the eight conditions evaluated in this research showed particles separated and with a good degree of sphericity, while in the other two particles had amorphous morphology and aggregation. In all treatments, most of the particles showed smooth surfaces. The average size of smallest particle obtained was 492 nm, while EDS results showed an even distribution of Ca2+ in the polymer matrix. The largest concentration of Ca2+ in ICP was 10.5%, which agrees with the theoretical value calculated, while the release kinetics showed an upward trend within 24 h. Using the technique employed in this research, it was possible to obtain nanoparticles loaded with calcium, of good size, sphericity and with release controlled properties. The characteristics of nanoparticles resulted from manipulation of the conditions of synthesis which allow control of the size and shape of the particles, and provides the means to adapt the properties of the materials to an specific application.

Keywords: calcium, controlled release, gelatin, nano spraydryer, nanofertilizer

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Authors: Kiran Nawaz, Ahmad Ali Shahid, Sehrish Iftikhar, Waheed Anwar, Muhammad Nasir Subhani

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Chili (Solanum annum L.) attacks by many fungal pathogens, including members of Oomycetes which are responsible for root rot in different chili growing areas of the world. Oomycetes pathogens cause economic losses in different regions of the Pakistan. Most of the plant tissues, including roots, crowns, fruit, and leaves, are vulnerable to Phytophthora capsici. It is very difficult to manage the Phytophthora root rot of chili as many commercial varieties are tremendously vulnerable to P. capsici. The causal agent of the disease was isolated on corn meal agar (CMA) and identified on a morphological basis by using available taxonomic keys. The pathogen was also confirmed on the molecular basis through internal transcribed spacer region and with other molecular markers.The Blastn results showed 100% homology with already reported sequences of P. capsici in NCBI database. Most of the farmers have conventionally relied on foliar fungicide applications to control Phytophthora root rot in spite of their incomplete effectiveness. In this study, in vitro plate assay, seed soaking and foliar applications of 6 fungicides were evaluated against root rot of chili. In vitro assay revealed that significant inhibition of linear growth was obtained with Triflumizole at 7.0%, followed by Thiophanate methyl (8.9%), Etridiazole (6.0%), Propamocarb (5.9%) and 7.5% with Mefenoxam and Iprodione for P. capsici. The promising treatments of in vitro plate bioassay were evaluated in pot experiments under controlled conditions in the greenhouse. All fungicides were applied after at 6-day intervals. Results of pot experiment showed that all treatments considerably inhibited the percentage of P. capsici root rot incidence. In addition, application of seed soaking with all six fungicides combined with the foliar spray of the same components showed the significant reduction in root rot incidence. The combine treatments of all fungicides as in vitro bioassay, seed soaking followed by foliar spray is considered non-harmful control methods which have advantages and limitation. Hence, these applications proved effective and harmless for the management of soil-borne plant pathogens.

Keywords: blastn, bioassay, corn meal agar(CMA), oomycetes

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Authors: K. Yu Vlasova, M. A. Abakumov, H. Wishwarsao, M. Sokolsky, N. V. Nukolova, A. G. Majouga, Y. I. Golovin, N. L. Klyachko, A. V. Kabanov

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Introduction:Nowadays pharmaceutical medicine is aimed to create systems for combined therapy, diagnostic, drug delivery and controlled release of active molecules to target cells. Magnetic nanoparticles (MNPs) are used to achieve this aim. MNPs can be applied in molecular diagnostics, magnetic resonance imaging (T1/T2 contrast agents), drug delivery, hyperthermia and could improve therapeutic effect of drugs. The most common drug containers, containing MNPs, are liposomes, micelles and polymeric molecules bonded to the MNPs surface. Usually superparamagnetic nanoparticles are used (the general diameter is about 5-6 nm) and all effects of high frequency magnetic field (MF) application are based on Neel relaxation resulting in heating of surrounded media. In this work we try to develop a new method to improve drug release from MNPs under super low frequency MF. We suppose that under low frequency MF exposures the Brown’s relaxation dominates and MNPs rotation could occur leading to conformation changes and release of bioactive molecules immobilized on MNPs surface.The aim of this work was to synthesize different systems with active drug (biopolymers coated MNPs nanoclusters with immobilized enzymes and doxorubicin (Dox) loaded magnetic liposomes/micelles) and investigate the effect of super low frequency MF on these drug containers. Methods: We have synthesized MNPs of magnetite with magnetic core diameter 7-12 nm . The MNPs were coated with block-copolymer of polylysine and polyethylene glycol. Superoxide dismutase 1 (SOD1) was electrostatically adsorbed on the surface of the clusters. Liposomes were prepared as follow: MNPs, phosphatidylcholine and cholesterol were dispersed in chloroform, dried to get film and then dispersed in distillated water, sonicated. Dox was added to the solution, pH was adjusted to 7.4 and excess of drug was removed by centrifugation through 3 kDa filters. Results: Polylysine coated MNPs formed nanosized clusters (as observed by TEM) with intensity average diameter of 112±5 nm and zeta potential 12±3 mV. After low frequency AC MF exposure we observed change of immobilized enzyme activity and hydrodynamic size of clusters. We suppose that the biomolecules (enzymes) are released from the MNPs surface followed with additional aggregation of complexes at the MF in medium. Centrifugation of the nanosuspension after AC MF exposures resulted in increase of positive charge of clusters and change in enzyme concentration in comparison with control sample without MF, thus confirming desorption of negatively charged enzyme from the positively charged surface of MNPs. Dox loaded magnetic liposomes had average diameter of 160±8 nm and polydispersity index (PDI) 0.25±0.07. Liposomes were stable in DW and PBS at pH=7.4 at 370C during a week. After MF application (10 min of exposure, 50 Hz, 230 mT) diameter of liposomes raised to 190±10 nm and PDI was 0.38±0.05. We explain this by destroying and/or reorganization of lipid bilayer, that leads to changes in release of drug in comparison with control without MF exposure. Conclusion: A new application of low frequency AC MF for drug delivery and controlled drug release was shown. Investigation was supported by RSF-14-13-00731 grant, K1-2014-022 grant.

Keywords: magnetic nanoparticles, low frequency magnetic field, drug delivery, controlled drug release

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Authors: Marcela Capcarova, Adriana Kolesarova, Marina Medvedova, Peter Petruska, Alexander V. Sirotkin

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The aim of this study was to determine the activity of superoxide dismutase (SOD), glutathione peroxidase (GPx), total antioxidant status (TAS) and accumulation of Hsp70 in porcine ovarian granulosa cells after deoxynivalenol (DON) and zearalenone (ZEA) exposure in vitro. Porcine ovarian granulosa cells were incubated with DON/ZEA administrations as follows: group A (10/10 ng/mL), group B (100/100 ng/mL), group C (1000/1000 ng/mL), and the control group without any additions for 24h. In this study mycotoxins developed stress reaction of porcine ovarian granulosa cells and increased accumulation of Hsp70 what resulted in increasing activities of SOD and GPx in groups with lower doses of mycotoxins. High dose of DON and ZEA had opposite effect on GPx activity than the lower doses. Slight increase in TAS of porcine granulosa cells was observed after mycotoxins exposure. These results contribute towards the understanding of cellular stress and its response.

Keywords: deoxynivalenol, zearalenone, antioxidants, Hsp70, granulosa cells

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Authors: Jessica Arthur, Duke Amegah, Kingsley Akenten Wiafe

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Background: Vanilla planifolia is the only edible fruit of the orchid family (Orchidaceae) among the over 35,000 Orchidaceae species found worldwide. In Ghana, Vanilla was discovered in the wild, but it is underutilized for commercial production, most likely due to a lack of knowledge on the best NAA and BAP combinations for in vitro propagation to promote successfully regenerated plant acclimatization. The growing interest and global demand for elite Vanilla planifolia plants and natural vanilla flavour emphasize the need for an effective industrial-scale micropropagation protocol. Tissue culture systems are increasingly used to grow disease-free plants and reliable in vitro methods can also produce plantlets with typically modest proliferation rates. This study sought to develop an efficient protocol for in vitro propagation of vanilla using nodal explants by testing different concentrations of NAA and BAP, for the proliferation of the entire plant. Methods: Nodal explants with dormant axillary buds were obtained from year-old laboratory-grown Vanilla planifolia plants. MS media was prepared with a nutrient stock solution (containing macronutrients, micronutrients, iron solution and vitamins) and semi-solidified using phytagel. It was supplemented with different concentrations of NAA and BAP to induce multiple shoots and roots (0.5mg/L BAP with NAA at 0, 0.5, 1, 1.5, 2.0mg/L and vice-versa). The explants were sterilized, cultured in labelled test tubes and incubated at 26°C ± 2°C with 16/8 hours light/dark cycle. Data on shoot and root growth, leaf number, node number, and survival percentage were collected over three consecutive two-week periods. The data were square root transformed and subjected to ANOVA and LSD at a 5% significance level using the R statistical package. Results: Shoots emerged at 8 days and roots at 12 days after inoculation with 94% survival rate. It was discovered that for the NAA treatments, MS media supplemented with 2.00 mg/l NAA resulted in the highest shoot length (10.45cm), maximum root number (1.51), maximum shoot number (1.47) and the highest number of leaves (1.29). MS medium containing 1.00 mg/l NAA produced the highest number of nodes (1.62) and root length (14.27cm). Also, a similar growth pattern for the BAP treatments was observed. MS medium supplemented with 1.50 mg/l BAP resulted in the highest shoot length (14.98 cm), the highest number of nodes (4.60), the highest number of leaves (1.75) and the maximum shoot number (1.57). MS medium containing 0.50 mg/l BAP and 1.0 mg/l BAP generated a maximum root number (1.44) and the highest root length (13.25cm), respectively. However, the best concentration combination for maximizing shoot and root was media containing 1.5mg/l BAP combined with 0.5mg/l NAA, and 1.0mg/l NAA combined with 0.5mg/l of BAP respectively. These concentrations were optimum for in vitro growth and production of Vanilla planifolia. Significance: This study presents a standardized protocol for labs to produce clean vanilla plantlets, enhancing cultivation in Ghana and beyond. It provides insights into Vanilla planifolia's growth patterns and hormone responses, aiding future research and cultivation.

Keywords: Vanilla planifolia, In vitro propagation, plant hormones, MS media

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Authors: Rania Abdulkareem Aboubakr Mahdaly Ammar

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The aortic valve (AV) is a complex mechanical environment that includes flexure, tension, pressure and shear stress forces to blood flow during cardiac cycle. This mechanical environment regulates AV tissue structure by constantly renewing and remodeling the phenotype. In vitro, ex vivo and in vivo studies have explained that pathological states such as hypertension and congenital defects like bicuspid AV ( BAV ) can potentially alter the AV’s mechanical environment, triggering a cascade of remodeling, inflammation and calcification activities in AV tissue. Changes in mechanical environments are first sent by the endothelium that induces changes in the extracellular matrix, and triggers cell differentiation and activation. However, the molecular mechanism of this process is not very well understood. Understanding these mechanisms is critical for the development of effective medical based therapies. Recently, there have been some interesting studies on characterizing the hemodynamics associated with AV, especially in pathologies like BAV, using different experimental and numerical methods. Here, we review the current knowledge of the local AV mechanical environment and its effect on valve biology, focusing on in vitro and ex vivo approaches.

Keywords: aortic valve mechanobiology, bicuspid calcification, pressure stretch, shear stress

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Authors: Monika Verma, Renuka Sharma, B. R. Gulati, Namita Singh

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In combination therapy, two chemotherapeutic agents work together in a collaborative action. It has appeared as one of the promising approaches to improve anti-cancer treatment efficacy. In the present investigation, piperine (P-NPS), paclitaxel (PTX NPS), and a combination of both, piperine-paclitaxel nanoparticle (Pip-PTX NPS), were made by the nanoprecipitation method and later characterized by PSA, DSC, SEM, TEM, and FTIR. All nanoparticles exhibited a monodispersed size distribution with a size of below 200 nm, zeta potential ranges from (-30-40mV) and a narrow polydispersity index (>0.3) of the drugs. The average encapsulation efficiency was found to be between 80 and 90%. In vitro release of drugs for nanoparticles was done spectrophotometrically. FTIR and DSC results confirmed the presence of the drug. The Pip-PTX NPS significantly inhibit cell proliferation as compared to the native drugs nanoparticles in the breast cancer cell line MCF-7. In addition, Pip-PTX NPS suppresses cells in colony formation and soft gel agar assay. Scratch migration and Transwell chamber invasion assays revealed that combined nanoparticles reduce the migration and invasion of breast cancer cells. Morphological studies showed that Pip-PTX NPS penetrates the cells and induces apoptosis, which was further confirmed by DNA fragmentation, SEM, and western blot analysis. Taken together, Pip-PTX NPS inhibits cell proliferation, anchorage dependent and anchorage independent cell growth, reduces migration and invasion, and induces apoptosis in cells. These findings support that combination therapy using Pip-PTX NPS represents a potential approach and could be helpful in the future for breast cancer therapy.

Keywords: piperine, paclitaxel, breast cancer, apoptosis

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Authors: Chuanxiang Cheng, Tiantian Min, Jin Yue

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Antimicrobial active packaging enables extensive biological effects to improve food safety. However, the efficacy of antimicrobial packaging hinges on factors including the diffusion rate of the active agent toward the food surface, the initial content in the antimicrobial agent, and the targeted food shelf life. Among the possibilities of antimicrobial packaging design, an interesting approach involves the incorporation of volatile antimicrobial agents into the packaging material. In this case, the necessity for direct contact between the active packaging material and the food surface is mitigated, as the antimicrobial agent exerts its action through the packaging headspace atmosphere towards the food surface. However, it still remains difficult to achieve controlled and precise release of bioactive compounds to the specific target location with required quantity in food packaging applications. Remarkably, the development of stimuli-responsive materials for electrospinning has introduced the possibility of achieving controlled release of active agents under specific conditions, thereby yielding enduring biological effects. Relative humidity (RH) for the storage of food categories such as meat and aquatic products typically exceeds 90%. Consequently, high RH can be used as an abiotic trigger for the release of active agents to prevent microbial growth. Hence, a novel RH - responsive polyvinyl alcohol/chitosan (PVA/CS) composite nanofibrous film incorporated with 4-terpineol/β-cyclodextrin inclusion complexes (4-TA@β-CD ICs) was engineered by electrospinning that can be deposited as a functional packaging materials. The characterization results showed the thermal stability of the films was enhanced after the incorporation due to the hydrogen bonds between ICs and polymers. Remarkably, the 4 wt% 4-TA@β-CD ICs/PVA/CS film exhibited enhanced crystallinity, moderate hydrophilic (Water contact angle of 81.53°), light barrier property (Transparency of 1.96%) and water resistance (Water vapor permeability of 3.17 g mm/m2 h kPa). Moreover, this film also showed optimized mechanical performance with a Young’s modulus of 11.33 MPa, a tensile strength of 19.99 MPa and an elongation at break of 4.44 %. Notably, the antioxidant and antibacterial properties of this packaging material were significantly improved. The film demonstrated the half-inhibitory concentrations (IC50) values of 87.74% and 85.11% for scavenging 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2, 2′-azinobis (3-ethylbenzothiazoline-6-sulfonic) (ABTS) free radicals, respectively, in addition to an inhibition efficiency of 65% against Shewanella putrefaciens, the characteristic bacteria in aquatic products. Most importantly, the film achieved controlled release of 4-TA under high 98% RH by inducing the plasticization of polymers caused by water molecules, swelling of polymer chains, and destruction of hydrogen bonds within the cyclodextrin inclusion complex. Consequently, low relative humidity is suitable for the preservation of nanofibrous film, while high humidity conditions typical in fresh food packaging environments effectively stimulated the release of active compounds in the film. This film with a long-term antimicrobial effect successfully extended the shelf life of Litopenaeus vannamei shrimp to 7 days at 4 °C. This attractive design could pave the way for the development of new food packaging materials.

Keywords: controlled release, electrospinning, nanofibrous film, relative humidity–responsive, shrimp preservation

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Authors: David Polcari, Samuel C. Perry, Loredano Pollegioni, Matthias Geissler, Janine Mauzeroll

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ᴅ-serine acts as an endogenous co-agonist for N-methyl-ᴅ-aspartate receptors in neuronal synapses. This makes it a key component in the development and function of a healthy brain, especially given its role in several neurodegenerative diseases such as Alzheimer’s disease and dementia. Despite such clear research motivations, the primary site and mechanism of ᴅ-serine release is still currently unclear. For this reason, we are developing a biosensor for the detection of ᴅ-serine utilizing a microelectrode in combination with a ᴅ-amino acid oxidase enzyme, which produces stoichiometric quantities of hydrogen peroxide in response to ᴅ-serine. For the fabrication of a biosensor with good selectivity, we use a permselective poly(meta-phenylenediamine) film to ensure only the target molecule is reacted, according to the size exclusion principle. In this work, we investigated the effect of the electrodeposition conditions used on the biosensor’s response time and selectivity. Careful optimization of the fabrication process allowed for enhanced biosensor response time. This allowed for the real time sensing of ᴅ-serine in a bulk solution, and also provided in means to map the efflux of ᴅ-serine in real time. This was done using scanning electrochemical microscopy (SECM) with the optimized biosensor to measure localized release of ᴅ-serine from an agar filled glass capillary sealed in an epoxy puck, which acted as a model system. The SECM area scan simultaneously provided information regarding the rate of ᴅ-serine flux from the model substrate, as well as the size of the substrate itself. This SECM methodology, which provides high spatial and temporal resolution, could be useful to investigate the primary site and mechanism of ᴅ-serine release in other biological samples.

Keywords: ᴅ-serine, enzymatic biosensor, microelectrode, scanning electrochemical microscopy

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Authors: Elhamalsadat Ghaffari, Moghan Amirhosseinian, Amir Khaleghipour

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Multifunctional bioactive glasses (BGs) are designed with a focus on the provision of bactericidal and biological properties desired for angiogenesis, osteogenesis, and ultimately potential applications in bone tissue engineering. To achieve these, six sol-gel copper/magnesium substituted derivatives of 58S-BG, i.e. a mol% series of 60SiO₂-4P₂O₅-5CuO-(31-x) CaO/xMgO (where x=0, 1, 3, 5, 8, and 10), were synthesized. Afterwards, the effect of MgO/CaO substitution on the in vitro formation of nano-hydroxyapatite (HA), osteoblast-like cell responses and BGs antibacterial performance were studied. During the BGs synthesis, the elimination of nitrates was achieved at 700 °C that prevented the BGs crystallization and stabilized the obtained dried gels. The structural and morphological evaluations were performed with X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), and scanning electron microscopy (SEM). These characterizations revealed that Cu-substituted 58S-BG consisting of 5 mol% MgO (BG-5/5) slightly had retarded the formation of HA. In addition, Cu-substituted 58S-BGs consisting 8 mol% and 10 mol% MgO (BG-5/8 and BG-5/10) displayed lower bioactivity probably due to the lower ion release rate of Ca–Si into the simulated body fluid (SBF). The determination of 3-(4, 5 dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) and alkaline phosphate (ALP) activities proved that the highest values of both differentiation and proliferation of MC3T3-E1 cells can be obtained from a 5 mol% MgO substituted BG, while the over addition of MgO (8 mol% and 10 mol%) decreased the bioactivity. Furthermore, these novel Cu/Mg-substituted 58S-BGs displayed antibacterial effect against methicillin-resistant Staphylococcus aureus bacteria. Taken together, the results suggest the equally-substituted BG-5/5 (i.e. the one consists of 5 mol% of both CuO and MgO) as a promising candidate for bone tissue engineering, among all newly designed BGs in this work, owing to its desirable cell proliferation, ALP activity and antibacterial properties.

Keywords: apatite, bioactivity, biomedical applications, sol-gel processes

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Authors: H. Maachou, A. Chagnes, G. Cote

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The present work reports the properties of chitosan/hydroxyapatite (Cs/HA: 100/00, 70/30 and 30/70) composite microspheres obtained by emulsification processing route. The morphology of chitosane microspheres was observed by a scanning electron microscope (SEM) which shows an aggregate of spherical microspheres with a particle size, determined by optical microscope, ranged from 4 to 10 µm. Thereafter, a biomimetic approach was used to study the in vitro biomineralization of these composites. It concerns the composites immersion in simulated body fluid (SBF) for different times. The deposited calcium phosphate was studied using X-ray diffraction analysis (XRD), FTIR spectroscopy and ICP analysis of phosphorus. In fact, the mineral formed on Cs/HA microspheres was a mixture of carbonated HA and β-TCP as showed by FTIR peaks at 1419,5 and 871,8 cm-1 and XRD peak at 29,5°. This formation was induced by the presence of HA in chitosan microspheres. These results are confirmed by SEM micrographs which chow the Ca-P crystals growth in form of cauliflowers. So, these materials are of great interest for bone regeneration applications due to their ability to nucleate calcium phosphates in presence of simulated body fluid (SBF).

Keywords: hydroxyapatite, chitosan, microsphere, composite, bone regeneration

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Authors: Abhay Asthana, Shally Toshkhani, Gyati Shilakari

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The present research was aimed to develop a biodegradable dermal patch formulation for wound healing in a novel, sustained and systematic manner. The goal is to reduce the frequency of dressings with improved drug delivery and thereby enhance therapeutic performance. In present study optimized formulation was designed using component polymers and excipients (e.g. Hydroxypropyl methyl cellulose, Ethylcellulose, and Gelatin) to impart significant folding endurance, elasticity and strength. Gelatin was used to get a mixture using ethylene glycol. Chitosan dissolved in suitable medium was mixed with stirring to gelatin mixture. With continued stirring to the mixture Curcumin was added in optimized ratio to get homogeneous dispersion. Polymers were dispersed with stirring in final formulation. The mixture was sonicated casted to get the film form. All steps were carried out under under strict aseptic conditions. The final formulation was a thin uniformly smooth textured film with dark brown-yellow color. The film was found to have folding endurance was around 20 to 21 times without a crack in an optimized formulation at RT (23C). The drug content was in range 96 to 102% and it passed the content uniform test. The final moisture content of the optimized formulation film was NMT 9.0%. The films passed stability study conducted at refrigerated conditions (4±0.2C) and at room temperature (23 ± 2C) for 30 days. Further, the drug content and texture remained undisturbed with stability study conducted at RT 23±2C for 45 and 90 days. Percentage cumulative drug release was found to be 80% in 12 h and matched the biodegradation rate as drug release with correlation factor R2 > 0.9. The film based formulation developed shows promising results in terms of stability and release profiles.

Keywords: biodegradable, patch, bioactive, polymer

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Authors: Elbadawy A. Kamoun

Abstract:

Magnetically responsive hydrogel nanocomposite (NCH) based on composites of superparamagnetic of Fe3O4 nano-particles and temperature responsive hydrogel matrices were developed. The nanocomposite hydrogel system based on the temperature sensitive N-isopropylacrylamide hydrogels crosslinked by poly(ethylene glycol)-400 dimethacrylate (PEG400DMA) incorporating with chitosan derivative, was synthesized and characterized. Likewise, the NCH system was synthesized by visible-light free radical photopolymerization, using carboxylated camphorquinone-amine system to avoid the common risks of the use of UV-light especially in hyperthermia treatment. Superparamagnetic of iron oxide nanoparticles were introduced into the hydrogel system by polymerizing mixture technique and monomer solution. FT-IR with Raman spectroscopy and Wide angle-XRD analysis were utilized to verify the chemical structure of NCH and exfoliation reaction for nanoparticles, respectively. Additionally, morphological structure of NCH was investigated using SEM and TEM photographs. The swelling responsive of the current nanocomposite hydrogel system with different crosslinking conditions, temperature, magnetic field efficiency, and the presence effect of magnetic nanoparticles were evaluated. Notably, hydrolytic degradation of this system was proved in vitro application. While, in-vivo release profile behavior is under investigation nowadays. Moreover, the compatibility and cytotoxicity tests were previously investigated in our studies for photoinitiating system. These systems show promised polymeric material candidate devices and are expected to have a wide applicability in various biomedical applications as mildly.

Keywords: hydrogel nanocomposites, tempretaure-responsive hydrogel, superparamagnetic nanoparticles, hyperthermia therapy

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Authors: Varsha Salian, Shama Rao, N. Narendra, B. Mohana Kumar

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Evolving evidence proposes the existence of a highly tumorigenic subpopulation of undifferentiated, self-renewing cancer stem cells, responsible for exhibiting resistance to conventional anti-cancer therapy, recurrence, metastasis and heterogeneous tumor formation. Importantly, the mechanisms exploited by cancer stem cells to resist chemotherapy are very less understood. Oral squamous cell carcinoma (OSCC) is one of the most regularly diagnosed cancer types in India and is associated commonly with alcohol and tobacco use. Therefore, the isolation and in vitro characterization of cancer stem-like cells from patients with OSCC is a critical step to advance the understanding of the chemoresistance processes and for designing therapeutic strategies. With this, the present study aimed to establish and characterize cancer stem-like cells in vitro from OSCC. The primary cultures of cancer stem-like cell lines were established from the tissue biopsies of patients with clinical evidence of an ulceroproliferative lesion and histopathological confirmation of OSCC. The viability of cells assessed by trypan blue exclusion assay showed more than 95% at passage 1 (P1), P2 and P3. Replication rate was performed by plating cells in 12-well plate and counting them at various time points of culture. Cells had a more marked proliferative activity and the average doubling time was less than 20 hrs. After being cultured for 10 to 14 days, cancer stem-like cells gradually aggregated and formed sphere-like bodies. More spheroid bodies were observed when cultured in DMEM/F-12 under low serum conditions. Interestingly, cells with higher proliferative activity had a tendency to form more sphere-like bodies. Expression of specific markers, including membrane proteins or cell enzymes, such as CD24, CD29, CD44, CD133, and aldehyde dehydrogenase 1 (ALDH1) is being explored for further characterization of cancer stem-like cells. To summarize the findings, the establishment of OSCC derived cancer stem-like cells may provide scope for better understanding the cause for recurrence and metastasis in oral epithelial malignancies. Particularly, identification and characterization studies on cancer stem-like cells in Indian population seem to be lacking thus provoking the need for such studies in a population where alcohol consumption and tobacco chewing are major risk habits.

Keywords: cancer stem-like cells, characterization, in vitro, oral squamous cell carcinoma

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Authors: Hairin Taha, Aditya Arya, M. A. Hapipah, A. M. Mustafa

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Plants have been an important source of medicine since ancient times. Pseuduvaria monticola is a rare montane forest species from the Annonaceae family. Traditionally, the plant was used to cure symptoms of fever, inflammation, stomach-ache and also to reduce the elevated levels of blood glucose. Scientifically, we have evaluated the antidiabetic potential of the Pseuduvaria monticola bark methanolic extract on certain in vitro cell based assays, followed by in vivo study. Results from in vitro models displayed PMm upregulated glucose uptake and insulin secretion in mouse pancreatic β-cells. In vivo study demonstrated the PMm down-regulated hyperglycaemia, oxidative stress and elevated levels of pro-inflammatory cytokines in type 2 diabetic rat models. Altogether, the study revealed that Pseuduvaria monticola might be used as a potential candidate for the management of type 2 diabetes and its related complications.

Keywords: type 2 diabetes, Pseuduvaria monticola, insulin secretion, glucose uptake

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Authors: Sina Gharevali

Abstract:

Staphylococcus aureus is one of the most important factors for nosocomial infections and infections acquired in a hospital setting role as is. Drug-resistant bacteria methicillin, which in 1961 was reported in many parts of the world, Made the role as the last drug, vancomycin, in the treatment of infections caused by the Staphylococcus aureus chain be taken into consideration. The aim of this study was to evaluate the antimicrobial effects of copper nanoparticles and compared it with antibiotics on Staphylococcus aureus resistant to vancomycin in vitro and animal model. In this study, this test was performed, and the most effective antibiotic for vancomycin-resistant Staphylococcus aureus was determined by disk diffusion method. After various concentrations of copper nanoparticles and antibiotics were prepared and vancomycin resistant Staphylococcus aureus bacteria with serial dilution method for determining antibiotic ciprofloxacin. Minimum Inhibitory Concentration and Minimum Bactericidal Concentrationcopper nanoparticles was performed. The agar dilution method for bacterial growth in different concentrations of copper nanoparticles and antibiotics ciprofloxacin was performed. The agar dilution method for bacterial growth in different concentrations of copper nanoparticles and antibiotics ciprofloxacin was performed. Then the broth dilution method for the antibiotic ciprofloxacin, nano-particles, and nano-particles of copper and copper-established antibiotic synergy MIC and MBC were obtained. MBC was obtained from the experimental animal model test method, and the results were compared. The results showed that copper nanoparticles compared with the antibiotic ciprofloxacin in vitro and animal model more effective in inhibiting the growth of Staphylococcus aureus resistant to vancomycin and ciprofloxacin and extent of the impact of the Synthetic effect of lower copper nanoparticles. Which can then be used to treat clinical research as a candidate.

Keywords: nanoparticles, copper, staphylococcus, aureus

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Authors: Jalil Badamfirooz

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Soil wind erosion has a negative impact on the environment. Mulching is one of the most efficient soil protection techniques. Bitumen emulsion has recently been utilized as a soil cover that is sprayed directly over the soil and forms a thin film. The thin coating of bitumen emulsion prevents soil erosion and keeps moisture in the soil. Besides, some compounds release into the soil and cause environmental problems. In the present study, the effect of bitumen emulsion on the release of polycyclic aromatic hydrocarbons (PAHs) into the soil is studied in an arid land located in the central part of Iran. The soil was Loamy-Sand and saline with a pH of 8.03. Bitumen emulsion was used in this study as mulch at a rate of 4 L m2. The effect of this mulch on soil properties was investigated after 6 months of mulch application. Then PAHs concentrations were determined in samples collected from different depths in bitumen emulsion sprayed and control soils. In general, bitumen emulsion application on soil led to a significant increase in some PAHs, which was higher than soil pollution standards critical level of pollution for commerce, groundwater protection, pasture forest, and park and residence uses.

Keywords: mulch, bitumen emulsion, arid land, PAH

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Authors: Pamita Awasthi, Kirna, Shilpa Dogra, Manu Vatsal, Ritu Barthwal

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Doxorubicin, also known as adriamycin, is an anthracycline class of drug used in cancer chemotherapy. It is used in the treatment of non-Hodgkin’s lymphoma, multiple myeloma, acute leukemias, breast cancer, lung cancer, endometrium cancer and ovary cancers. It functions via intercalating DNA and ultimately killing cancer cells. The major side effects of doxorubicin are hair loss, myelosuppression, nausea & vomiting, oesophagitis, diarrhoea, heart damage and liver dysfunction. The minor modifications in the structure of compound exhibit large variation in the biological activity, has prompted us to carry out the synthesis of sulfonamide derivatives. Sulfonamide is an important feature with broad spectrum of biological activity such as antiviral, antifungal, diuretics, anti-inflammatory, antibacterial and anticancer activities. Structure of the synthesized compound N-(1-methyl-2-oxo-2-N-methyl anilino-ethyl)benzene sulfonamide confirmed by proton nuclear magnetic resonance (1H NMR),13C NMR, Mass and FTIR spectroscopic tools to assure the position of all protons and hence stereochemistry of the molecule. Further we have reported the binding potential of synthesized sulfonamide analogues in comparison to doxorubicin drug using Auto Dock 4.2 software. Computational binding energy (B.E.) and inhibitory constant (Ki) has been evaluated for the synthesized compound in comparison of doxorubicin against Poly (dA-dT).Poly (dA-dT) and Poly (dG-dC).Poly (dG-dC) sequences. The in vitro cytotoxic study against human breast cancer cell lines confirms the better anticancer activity of the synthesized compound over currently in use anticancer drug doxorubicin. The IC50 value of the synthesized compound is 7.12 µM where as for doxorubicin is 7.2 µ.

Keywords: Doxorubicin, auto dock, in silco, in vitro

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Authors: Ming Chian Yew, Ming Kun Yew, Lip Huat Saw, Tan Ching Ng, Rajkumar Durairaj, Jing Han Beh

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This paper analyzes the fire protection performance, char formation and heat release characteristics of the thin film intumescent coatings that incorporate waste eggshell (ES) as a nano bio-filler. In this study, the Bunsen burner and the fire propagation (BS 476: Part 6) tests of coatings were measured. Experiments on the samples were also tested to evaluate their fire behavior using a cone calorimeter according to ISO 5660-1 specifications. On exposure, the samples B, C and D had been certified to be Class 0 due to the fire propagation indexes of the samples were less than 12. Samples B and D showed a significant reduction in total heat rate (B=11.6 MJ/m² and D=12.0 MJ/m²) and uniform char structures with the addition of 3.30 wt.% and 2.75 wt.% ES nano bio-filler, respectively. As a result, ES nano bio-filler composition good to slow down the fire expanding and demonstrate better fire protection due to its positive synergistic effect with flame retardant ingredients on physical and chemical reactions in fire protection.

Keywords: cone calorimeter, eggshell, fire protection, heat release rate, intumescent coating

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Authors: Kotchamon Yodkhum, Thawatchai Phaechamud

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Hydrophobic chitosan-based materials have been developed as controlled drug delivery system. This study was aimed to prepare and evaluate chitosan-aluminum monostearate composite dispersion (CLA) as fabricating liquid for construct a hydrophobic, controlled-release solid drug delivery matrix. This work was attempted to blend hydrophobic substance, aluminum monostearate (AMS), with chitosan in acidic aqueous medium without using any surfactants or grafting reaction, and high temperature during mixing that are normally performed when preparing hydrophobic chitosan system. Lactic acid solution (2%w/v) was employed as chitosan solvent. CLA dispersion was prepared by dispersing different amounts of AMS (1-20% w/w) in chitosan solution (4% w/w) with continuous agitation using magnetic stirrer for 24 h. Effect of AMS amount on physicochemical properties of the dispersion such as viscosity, rheology and particle size was evaluated. Morphology of chitosan-AMS complex (dispersant) was observed under inverted microscope and atomic force microscope. Stability of CLA dispersions was evaluated after preparation within 48 h. CLA dispersions containing AMS less than 5 % w/w exhibited rheological behavior as Newtonian while that containing higher AMS amount exhibited as pseudoplastic. Particle size of the dispersant was significantly smaller when AMS amount was increased up to 5% w/w and was not different between the higher AMS amount system. Morphology of the dispersant under inverted microscope displayed irregular shape and their size exhibited the same trend with particle size measurement. Observation of the dispersion stability revealed that phase separation occurred faster in the system containing higher AMS amount which indicated lower stability of the system. However, the dispersions were homogeneous and stable more than 12 hours after preparation that enough for fabrication process. The prepared dispersions had ability to be fabricated as a porous matrix via lyophilization technique.

Keywords: chitosan, aluminum monostearate, dispersion, controlled-release

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Authors: Malek Y. S. Ibrahim, Jeffrey A. Bennett, Milad Abolhasani

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Despite the potential of hydrogen (H2) storage in liquid organic carriers to achieve carbon neutrality, the energy required for H2 release and the cost of catalyst recycling has hindered its large-scale adoption. In response, a photo flow reactor packed with rhodium (Rh)/titania (TiO2) photocatalyst was reported for the continuous and selective acceptorless dehydrogenation of 1,2,3,4-tetrahydroquinoline to H2 gas and quinoline under visible light irradiation at room temperature. The tradeoff between the reactor pressure drop and its photocatalytic surface area was resolved by selective in-situ photodeposition of Rh in the photo flow reactor post-packing on the outer surface of the TiO2 microparticles available to photon flux, thereby reducing the optimal Rh loading by 10 times compared to a batch reactor, while facilitating catalyst reuse and regeneration. An example of using quinoline as a hydrogen acceptor to lower the energy of the hydrogen production step was demonstrated via the water-gas shift reaction.

Keywords: hydrogen storage, flow chemistry, photocatalysis, solar hydrogen

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Authors: Davender Kumar

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Aim: It is important for the orthodontist to be familiar with the sliding resistance (SR) generated by the ligation method used during the space closure phase with sliding mechanics. To determine new, experimental non-conventional (slide) ligature demonstrates less friction in vitro when compared other ligatures on the market. Methods: Experimental in vitro were carried out to test the performance of the low-friction system with regard to assess the forces released by different bracket–ligature systems with bonded in iron plate mounted on an Instron machine. Results: The outcomes of experimental testing showed that the combination of the low-friction ligatures with the super elastic nickel-titanium and SS wires produced a significantly smaller amount of binding at the bracket/arch wire/ligature unit when compared to conventional elastomeric ligatures. Conclusion: The biomechanical consequences of the use of low-friction ligatures were shorter duration of orthodontic treatment during the levelling and aligning phase, concurrent dentoalveolar expansion of the dental arch, and the possibility of using biologically adequate orthodontic forces.

Keywords: archwire, bracket, friction, ligation

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