Search results for: in vitro cultures
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 2261

Search results for: in vitro cultures

731 Biodegradation of 2,4-Dichlorophenol by Pseudomonas chlororaphis Strain Isolated from Activated Sludge Sample from a Wastewater Treatment Plant in Durban, South Africa

Authors: Boitumelo Setlhare, Mduduzi P. Mokoena, Ademola O. Olaniran

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Agricultural and industrial activities have led to increasing production of xenobiotics such as 2,4-dichlorophenol (2,4-DCP), a derivative of 2,4-dichlorophenoxyacetic acid (2,4-D), which is a widely used herbicide. Bioremediation offers an efficient, cost-effective and environmentally friendly method for degradation of the compound through the activities of the various microbial enzymes involved in the catabolic pathway. The aim of this study was to isolate and characterize bacterial isolate indigenous to contaminated sites in Durban, South Africa for 2,4-DCP degradation. One bacterium capable of utilizing 2,4-DCP as sole carbon source was isolated using culture enrichment technique and identified as Pseudomonas chlororaphis strain UFB2 via PCR amplification and analysis of 16S rRNA gene sequence. This isolate was able to degrade up to 75.11% of 2,4-DCP in batch cultures within 10 days, with the degradation rate constant of 0.14 mg/l/d. Phylogenetic analysis revealed the relatedness of this bacterial isolate to other Pseudomonas sp. previously characterized for chlorophenol degradation. PCR amplification of the catabolic genes involved in 2,4-DCP degradation revealed the presence of the correct amplicons for phenol hydroxylase (600 bp), catechol 1,2-dioxygenase (214 bp), muconate isomerase (851 bp), cis-dienelactone hydrolase (577 bp), and trans-dienelactone hydrolase (491 bp) genes. Enzyme assays revealed activity as high as 21840 mU/mg, 15630 mU/mg, 2340 mU/mg and 1490 mU/mg obtained for phenol hydroxylase, catechol 1,2-dioxygenase, cis-dienelactone hydroxylase and trans-dienelactone hydroxylase, respectively. The absence of catechol 2,3-dioxygenase gene and the corresponding enzyme in this isolate suggests that the organism followed ortho-pathway for 2,4-DCP degradation. Furthermore, the absence of malaycetate reductase genes showed that the bacterium may not be able to completely mineralize 2,4-DCP. Further studies are required to optimize 2,4-DCP degradation by this isolate as well as to elucidate the mechanism of 2,4-DCP degradation.

Keywords: biodegradation, catechol 1, 2-dioxygenase, 2, 4-dichlorophenol, phenol hydroxylase, Pseudomonas chlororaphis

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730 Adopting a Comparative Cultural Studies Approach to Teaching Writing in the Global Classroom

Authors: Madhura Bandyopadhyay

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Teaching writing within multicultural and multiethnic communities poses many unique challenges not the least of which is that of intercultural communication. When the writing is in English, pedagogical imperatives often encounter the universalizing tendencies of standardization of both language use and structural parameters which are often at odds with maintaining local practices which preserve cultural pluralism. English often becomes the contact zone within which individual identities of students play out against the standardization imperatives of the larger world. Writing classes can serve as places which become instruments of assimilation of ethnic minorities to a larger globalizing or nationalistic agenda. Hence, for those outside of the standard practices of writing English, adaptability towards a mastery of those practices valued as standard become the focus of teaching taking away from diversity of local English use and other modes of critical thinking. In a very multicultural and multiethnic context such as the US or Singapore, these dynamics become very important. This paper will argue that multiethnic writing classrooms can greatly benefit from taking up a cultural studies approach whereby the students’ lived environments and experiences are analyzed as cultural texts to produce writing. Such an approach eliminates limitations of using both literary texts as foci of discussion as in traditional approaches to teaching writing and the current trend in teaching composition without using texts at all. By bringing in students’ lived experiences into the classroom and analyzing them as cultural compositions stressing the ability to communicate across cultures, cultural competency is valued rather than adaptability while privileging pluralistic experiences as valuable even as universal shared experience are found. Specifically, while teaching writing in English in a multicultural classroom, a cultural studies approach makes both teacher and student aware of the diversity of the English language as it exists in our global context in the students’ experience while making space for diversity in critical thinking, structure and organization of writing effective in an intercultural context.

Keywords: English, multicultural, teaching, writing

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729 Mechanisms and Regulation of the Bi-directional Motility of Mitotic Kinesin Nano-motors

Authors: Larisa Gheber

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Mitosis is an essential process by which duplicated genetic information is transmitted from mother to daughter cells. Incorrect chromosome segregation during mitosis can lead to genetic diseases, chromosome instability and cancer. This process is mediated by a dynamic microtubule-based intracellular structure, the mitotic spindle. One of the major factors that govern the mitotic spindle dynamics are the kinesin-5 biological nano motors that were believed to move unidirectionally on the microtubule filaments, using ATP hydrolysis, thus performing essential functions in mitotic spindle dynamics. Surprisingly, several reports from our and other laboratories have demonstrated that some kinesin-5 motors are bi-directional: they move in minus-end direction on the microtubules as single-molecules and can switch directionality under a number of conditions. These findings broke a twenty-five-years old dogma regarding kinesin directionality (1, 2). The mechanism of this bi-directional motility and its physiological significance remain unclear. To address this unresolved problem, we apply an interdisciplinary approach combining live cell imaging, biophysical single molecule, and structural experiments to examine the activity of these motors and their mutated variants in vivo and in vitro. Our data shows that factors such as protein phosphorylation (3, 4), motor clustering on the microtubules (5, 6) and structural elements (7, 8) regulate the bi-directional motility of kinesin motors. We also show, using Cryo-EM, that bi-directional kinesin motors obtain non-canonical microtubule binding, which is essential to their special motile properties and intracellular functions. We will discuss the implication of these findings to mechanism bi-directional motility and physiological roles in mitosis.

Keywords: mitosis, cancer, kinesin, microtubules, biochemistry, biophysics

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728 Isolation, Characterization, and Antibacterial Activity of Endophytic Bacteria from Iranian Medicinal Plants

Authors: Maryam Beiranvand, Sajad Yaghoubi

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Background: Some microbes can colonize plants’ inner tissues without causing obvious damage and can even produce useful bioactive substances. In the present study, the diversity of the endophytic bacteria associated with medicinal plants from Iran was investigated by culturing techniques, molecular gene identification, as well as measuring them for antibacterial activity. Results: In the spring season from 2013 to 2014, 35 herb pharmacology samples were collected, sterilized, meshed, and then cultured on selective media culture. A total of 199 endophytic bacteria were successfully isolated from 35 tissue cultures of medical plants, and sixty-seven out of 199 bacterial isolates were subjected to identification by the 16S rRNA gene sequence analysis method. Based on the sequence similarity gene and phylogenetic analyses, these isolates were grouped into five classes, fourteen orders, seventeen families, twenty-one genera, and forty strains. The most abundant group of endophytic bacteria was actinobacterial, consisting of thirty-two (47%) out of 67 bacterial isolates. Ten (22.3%) out of 67 bacterial isolates remained unidentified and classified at the genus level. The signature of the 16S rRNA gene formed a distinct line in a phylogenetic tree showing that they might be new species of bacteria. One (5.2%) out of 67 bacterial isolates was still not well categorized. Forty-two out of 67 strains were candidates for antimicrobial activity tests. Nineteen (45%) out of 42 strains showed antimicrobial activity multidrug-resistance (MDR); thirteen (68%) out of 19 strains were allocated to classes actinobacteria. Four (21%) out of 19 strains belonged to the Bacillaceae family, one (5.2%) out of 19 strains was the Paenibacillaceae family, and one (5.2%) out of 19 strains belonged to the Pseudomonadaceae family. The other twenty-three strains did not show inhibitory activities. Conclusions: Our research showed a high-level phylogenetic diversity and the intoxicating antibiotic activity of endophytic bacteria in the herb pharmacology of Iran.

Keywords: medical plant, endophytic bacteria, antimicrobial activity, whole genome sequencing analysis

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727 Preparation and Characterization of Chitosan Nanoparticles for Delivery of Oligonucleotides

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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726 Biological Treatment of Corn Stover with Pleurotus ostreatus, Pleurotus eryngii and Lentinula edudes to Improve Digestibility

Authors: Aydan Atalar, Nurcan Cetinkaya

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Corn stover is leftover of the leaves, stalk, husks and tassels in the field after harvesting the grain combined. Corn stover is a low-quality roughage but has mostly been used as roughage source for feeding ruminant animals in developing countries including Turkey; however, it can also be used to make biofuels as in developed countries. The objectives of the present study were to improve the digestibility of corn stover by the treatment of white rod fungus mainly Pleurotus osteritus (PO), Pleurotus eryingii (PE) and Lantinula edudes (LE) at different incubation times and also to determine the most effective fungus and incubation time to prepare fermeted corn stover for ruminant nutrition. The choped corn stover was treated with PO, PE and LE and incubated for 10, 20, 30 and 40 days in incubator at 26 0C. After each incubation time dry matter(DM), organic matter(OM), crude protein (CP), neutral detergent fiber (NDF), acid detergent fiber (ADF), neutral detergent lignin (ADL), in-vitro true dry matter digestibility (IVTDMD) and organic matter digestibility (IVTOMD) were determined. The mean IVTDMD and IVTOMD levels were increased by PO, PE and LE treatments in increasing order of incubation times. The obtained IVTDM values were 59.45, 60.51, 60.82 and 60.18 %; 59.45, 70.55, 67.18 and 66.96 %; 59.45, 70.55, 67.18 and 66,96 %; 59.45, 74.90, 69.18 % ; 59.45, 76.50, 71.24 and 73.04 for control, PO, PE and LE treatments at 0, 10, 20, 30 and 40 days incubation times respectively. The obtained IVTOMD values were 56.45,60.26,60.82and 60.18 %; 56.45, 68.70, 67.18 and 66.96 %; 56.45, 71.26, 69.18 and 69.28 %; 56.45, 73.23, 71.24 and 73.04 % for control, PO, PE and LE treatments at 0, 10, 20, 30 and 40 days incubation times respectively. The most effective fungus was PO and the incubation time was 30 days. In conclusion, PO treatment of corn stover with 30 days incubation may be used to prepare fermented corn stover for ruminant nutrition.

Keywords: biological treatment, corn stover, digestibility, Lantinula edudes, Pleurotus eryingii, Pleurotus osteritus

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725 Anticancer Effect of Resveratrol-Loaded Gelatin Nanoparticles in NCI-H460 Non-Small Cell Lung Carcinoma Cell Lines

Authors: N. Rajendra Prasad

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Resveratrol (RSV), a grape phytochemical, has drawn greater attention because of its beneficial ef-fects against cancer. However, RSV has some draw-backs such as unstabilization, poor water solubility and short biological half time, which limit the utili-zation of RSV in medicine, food and pharmaceutical industries. In this study, we have encapsulated RSV in gelatin nanoparticles (GNPs) and studied its anti-cancer efficacy in NCI-H460 lung cancer cells. SEM and DLS studies have revealed that the prepared RSV-GNPs possess spherical shape with a mean diameter of 294 nm. The successful encapsulation of RSV in GNPs has been achieved by the cross-linker glutaraldehyde probably through Schiff base reaction and hydrogen bond interaction. Spectrophotometric analysis revealed that the max-imum of 93.6% of RSV has been entrapped in GNPs. In vitro drug release kinetics indicated that there was an initial burst release followed by a slow and sustained release of RSV from GNPs. The prepared RSV-GNPs exhibited very rapid and more efficient cellular uptake than free RSV. Further, RSV-GNPs treatment showed greater antiproliferative efficacy than free RSV treatment in NCI-H460 cells. It has been found that greater ROS generation, DNA damage and apoptotic incidence in RSV-GNPs treated cells than free RSV treatment. Erythrocyte aggregation assay showed that the prepared RSV-GNPs formulation elicit no toxic response. HPLC analysis revealed that RSV-GNPs was more bioavailable and had a longer half-life than free RSV. Hence, GNPs carrier system might be a promising mode for controlled delivery and for improved therapeutic index of poorly water soluble RSV.

Keywords: resveratrol, coacervation, anticancer gelatin nanoparticles, lung cancer, controlled release

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724 The Protective Role of Decoy Receptor 3 Analogue on Rat Steatotic Liver against Ischemia-Reperfusion Injury by Blocking M1/Th1 Polarization and Multiple Upstream Pathogenic Cascades

Authors: Tzu-Hao Li, Shie-Liang Hsieh, Han-Chieh Lin, Ying-Ying Yang

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TNF superfamily-stimulated pathogenic cascades and macrophage (M1)/kupffer cells (KC) polarization are important in the pathogenesis of ischemia-reperfusion (IR) liver injury in animals with hepatic steatosis (HS). Decoy receptor 3 (DcR3) is a common upstream inhibitor of the above-mentioned pathogenic cascades. The study evaluated whether modulation of these DcR3-related cascades was able to protect steatotic liver from IR injury. Serum and hepatic DcR3 levels were lower in patients and animals with HS. Accordingly, the effects of pharmacologic and genetic DcR3 replacement on the IR-related pathogenic changes were measured. Significantly, DcR3 replacement protected IR-Zucker(HS) rats and IR-DcR3-Tg(HS) mice from IR liver injury. The beneficial effects of DcR3 replacement were accompanied by decreased serum/hepatic TNF, soluble TNF-like cytokine 1A (TL1A), Fas ligand (Fas-L) and LIGHT, T-helper-cell-1 cytokine (INF) levels, neutrophil infiltration, M1 polarization, neutrophil-macrophage/KC-T-cell interaction, hepatocyte apoptosis and improved hepatic microcirculatory failure among animals with IR-injured steatotic livers. Additionally, TL1A, Fas-L, LIGHT and TLR4/NFB signals were found to mediate the DcR3-related protective effects of steatotic livers from IR injury. Using multimodal in vivo and in vitro approaches, we found that DcR3 was a potential agent to protect steatotic livers from IR injury by simultaneous blocking the multiple IR injury-related pathogenic changes.

Keywords: Decoy 3 receptor, ischemia-reperfusion injury, M1 polarization, TNF superfamily

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723 Wound Healing and Antioxidant Properties of 80% Methanol Leaf Extract of Verbascum sinaiticum (Scrophulariaceae), an Ethiopian Medicinal Plant

Authors: Solomon Assefa Huluka

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Wounds account for severe morbidity, socioeconomic distress, and mortality around the globe.For several years, various herbal products are used to expediteand augment the innate wound healing process. In Ethiopian folkloricmedicine, Verbascum sinaiticum L. (V. sinaiticum) is commonlyapplied as a wound-healing agent. The present study investigated the potential wound healing and antioxidant properties of hydroalcoholic leaf extract of V. sinaiticum. The 80% methanol extract, formulated as 5% (w/w) and 10% (w/w) ointments, was evaluated in excision and incision wound models using nitrofurazone and simple ointment as positive and negative controls, respectively. Parameters such as wound contraction, period of epithelialization, and tensile strength were determined. Moreover, its in vitro antioxidant property was evaluated using a DPPH assay. In the excision model, both doses (5% and 10% w/w) of the extract showed a significant (p<0.001) wound healing efficacy compared to the negative control, as evidenced by enhanced wound contraction rate and shorter epithelialization time records. In the incision model, the lower dose (5% w/w) ointment formulation of the extract exhibited the maximum increment in tensile strength (85.6%) that was significant (p<0.001)compared to negative and untreated controls. Animals treated with 5% w/wointment, furthermore, showed a significantly (p < 0.05) higher percentage of tensile strength than nitrofurazone treated ones. Moreover, the hydroalcoholic extract of the plant showed a noticeable free radical scavenging property. The result of the present study upholds the folkloric use of V. sinaiticum in the treatment of wounds.

Keywords: wound healing, antioxidant, excision wound model, incision wound model, verbascum sinaiticum

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722 CP-96345 Rregulates Hydrogen Sulphide Induced TLR4 Signaling Pathway Adhesion Molecules in Caerulein Treated Pancreatic Acinar Cells

Authors: Ramasamy Tamizhselvi, Leema George, Madhav Bhatia

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We have earlier shown that mouse pancreatic acinar cells produce hydrogen sulfide (H2S) and play a role in the pathogenesis of acute pancreatitis. This study is to determine the effect of H2S on TLR4 mediated innate immune signaling in acute pancreatitis via substance P (SP). Male Swiss mice were treated with hourly intraperitoneal injection of caerulein (50μg/kg) for 10 hour. DL-propargylglycine (PAG) (100 mg/kg i.p.), an inhibitor of H2S formation was administered 1h after the induction of acute pancreatitis. Pancreatic acinar cells from male Swiss mice were incubated with or without caerulein (10–7 M for 60 min) and CP-96345 (NK1R inhibitor). To better understand the effect of H2S in inflammation, acinar cells were stimulated with caerulein after addition of H2S donor, NaHS. In addition, caerulein treated pancreatic acinar cells were pretreated with PAG (30 µM), for 1h. H2S inhibitor, PAG, eliminated TLR4, IRAK4, TRAF6 and NF-kB levels in an in vitro and in vivo model of caerulein-induced acute pancreatitis. PPTA gene deletion reduced TLR4, MyD88, IRAK4, TRAF6, adhesion molecules and NF-kB in caerulein treated pancreatic acinar cells whereas administration of NaHS resulted in further rise in TLR4 and NF-kB levels in caerulein treated pancreatic acinar cells. In addition, acini isolated from mice and treated with PPTA gene receptor NK1R antagonist CP96345 did not exhibit further increase in TLR4, IRAK4, TRAF6, adhesion molecules and NF-kB levels after NaHS pretreatment. The present findings show for the first time that in acute pancreatitis, H2S up-regulates TLR4 pathway and NF-kB via substance P.

Keywords: preprotachykinin-A gene, H2S, TLR4, acute pancreatitis

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721 The Activity of Polish Propolis and Cannabidiol Oil Extracts on Glioblastoma Cell Lines

Authors: Sylwia K. Naliwajko, Renata Markiewicz-Zukowska, Justyna Moskwa, Krystyna Gromkowska-Kepka, Konrad Mielcarek, Patryk Nowakowski, Katarzyna Socha, Anna Puscion-Jakubik, Maria H. Borawska

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Glioblastoma (grade IV WHO) is a rapidly progressive brain tumor with very high morbidity and mortality. The vast malignant gliomas are not curable despite the therapy (surgical, radiotherapy, chemotherapy) and patients seek alternative or complementary treatments. Patients often use cannabidiol (CBD) oil as an alternative therapy of glioblastoma. CBD is one of the cannabinoids, an active component of Cannabis sativa. THC (Δ9-tetrahydrocannabinol) can be addictive, and in many countries CBD oil without THC ( < 0,2%) is available. Propolis produced by bees from the resin collected from trees has antiglioma properties in vitro and can be used as a supplement in complementary therapy of gliomas. The aim of this study was to examine the influence of extract from CBD oil in combination with propolis extract on two glioblastoma cell lines. The MTT (Thiazolyl Blue Tetrazolium Bromide) test was used to determine the influence of CBD oil extract and polish propolis extract (PPE) on the viability of glioblastoma cell lines – U87MG and LN18. The cells were incubated (24, 48 and 72 h) with CBD oil extract and PPE. CBD extract was used in concentration 1, 1.5 and 3 µM and PPE in 30 µg/mL. The data were presented compared to the control. The statistical analysis was performed using Statistica v. 13.0 software. CBD oil extract in concentrations 1, 1.5 and 3 µM did not inhibit the viability of U87MG and LN18 cells (viability more than 90% cells compared to the control). There was no dose-response viability, and IC50 value was not recognized. PPE in the concentration of 30 µg/mL time-dependently inhibited the viability of U87MG and LN18 cell line (after 48 h the viability as a percent of the control was 59,7±6% and 57,8±7%, respectively). In a combination of CBD with PPE, the viability of the treated cells was similar to PPE used alone (58,2±7% and 56,5±9%, respectively). CBD oil extract did not show anti-glioma activity and in combination with PPE did not change the activity of PPE.

Keywords: anticancer, cannabidiol, cell line, glioblastoma

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720 Reduced Glycaemic Impact by Kiwifruit-Based Carbohydrate Exchanges Depends on Both Available Carbohydrate and Non-Digestible Fruit Residue

Authors: S. Mishra, J. Monro, H. Edwards, J. Podd

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When a fruit such as kiwifruit is consumed its tissues are released from the physical /anatomical constraints existing in the fruit. During digestion they may expand several-fold to achieve a hydrated solids volume far greater than the original fruit, and occupy the available space in the gut, where they surround and interact with other food components. Within the cell wall dispersion, in vitro digestion of co-consumed carbohydrate, diffusion of digestion products, and mixing responsible for mass transfer of nutrients to the gut wall for absorption, were all retarded. All of the foregoing processes may be involved in the glycaemic response to carbohydrate foods consumed with kiwifruit, such as breakfast cereal. To examine their combined role in reducing the glycaemic response to wheat cereal consumed with kiwifruit we formulated diets containing equal amounts of breakfast cereal, with the addition of either kiwifruit, or sugars of the same composition and quantity as in kiwifruit. Therefore, the only difference between the diets was the presence of non-digestible fruit residues. The diet containing the entire disperse kiwifruit significantly reduced the glycaemic response amplitude and the area under the 0-120 min incremental blood glucose response curve (IAUC), compared with the equicarbohydrate diet containing the added kiwifruit sugars. It also slightly but significantly increased the 120-180 min IAUC by preventing a postprandial overcompensation, indicating improved homeostatic blood glucose control. In a subsequent study in which we used kiwifruit in a carbohydrate exchange format, in which the kiwifruit carbohydrate partially replaced breakfast cereal in equal carbohydrate meals, the blood glucose was further reduced without a loss of satiety, and with a reduction in insulin demand. The results show that kiwifruit may be a valuable component in low glycaemic impact diets.

Keywords: carbohydrate, digestion, glycaemic response, kiwifruit

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719 Smart Coating for Enhanced Corneal Healing via Delivering Progranulin

Authors: Dan Yan, Yunuo Zhang, Yuhan Huang, Weijie Ouyang

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The cornea serves as a vital protective barrier for the eye; however, it is prone to injury and damage that can disrupt corneal epithelium and nerves, triggering inflammation. Therefore, understanding the biological effects and molecular mechanisms involved in corneal wound healing and identifying drugs targeting these pathways is crucial for researchers in this field. This study aimed to investigate the therapeutic potential of progranulin (PGRN) in treating corneal injuries. Our findings demonstrated that PGRN significantly enhanced corneal wound repair by accelerating corneal re-epithelialization and re-innervation. In vitro experiments with cultured epithelial cells and trigeminal ganglion cells further revealed that PGRN stimulated corneal epithelial cell proliferation and promoted axon growth in trigeminal ganglion cells. Through RNA-sequencing (RNA-seq) analysis and other experimental techniques, we discovered that PGRN exerted its healing effects by modulating the Wnt signaling pathway, which played a critical role in repairing epithelial cells and promoting axon regeneration in trigeminal neurons. Importantly, our study highlighted the anti-inflammatory properties of PGRN by inhibiting the NF-κB signaling pathway, leading to decreased infiltration of macrophages. In conclusion, our findings underscored the potential of PGRN in facilitating corneal wound healing by promoting corneal epithelial cell proliferation, trigeminal ganglion cell axon regeneration, and suppressing ocular inflammation. These results suggest that PGRN could potentially expedite the healing process and improve visual outcomes in patients with corneal injuries.

Keywords: cornea, wound healing, progranulin, corneal epithelial cells, trigeminal ganglion cells

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718 Transcriptomic Response of Calmodulin Encoding Gene (CaM) in Pesticide Utilizing Talaromyces Fungal Strains

Authors: M. D. Asemoloye, S. G. Jonathan, A. Rafiq, O. J. Olawuyi, D. O. Adejoye

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Calmodulin is one of the intracellular calcium proteins that regulates large spectrum of enzymes and cellular functions including metabolism of cyclic nucleotides and glycogen. The potentials of calmodulin gene in fungi necessitates their genetic response and their strong cassette of enzyme secretions for pesticide degradation. Therefore, this study was carried out to investigate the ‘Transcriptomic’ response of calmodulin encoding genes in Talaromyces fungi in response to 2, 2-dichlorovinyl dimethyl phosphate (DDVP or Dichlorvos) an organophosphate pesticide and γ-Hexachlorocyclohexane (Lindane) an organochlorine pesticide. Fungi strains isolated from rhizosphere from grasses rhizosphere in pesticide polluted sites were subjected to percentage incidence test. Two most frequent fungi were further characterized using ITS gene amplification (ITS1 and ITS4 combinations), they were thereafter subjected to In-vitro DDVP and lindane tolerance tests at different concentrations. They were also screened for presence and expression of calmodulin gene (caM) using RT-PCR technique. The two Talaromyces strains had the highest incidence of 50-72% in pesticide polluted site, they were both identified as Talaromyces astroroseus asemoG and Talaromyces purpurogenum asemoN submitted in NCBI gene-bank with accession numbers KY488464 and KY488468 respectively. T. astroroseus KY488464 tolerated DDVP (1.23±0.023 cm) and lindane (1.11±0.018 cm) at 25 % concentration while T. purpurogenum KY488468 tolerated DDVP (1.33±0.061 cm) and lindane (1.54±0.077 cm) at this concentration. Calmodulin gene was detected in both strains, but RT-PCR expression of caM gene revealed at 900-1000 bp showed an under-expression of caM in T. astrorosues KY488464 but overexpressed in T. purpurogenum KY488464. Thus, the calmodulin gene response of these fungal strains to both pesticides could be considered in monitoring the potentials of fungal strains to pesticide tolerance and bioremediation of pesticide in polluted soil.

Keywords: Calmodulin gene, pesticide, RT-PCR, talaromyces, tolerance

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717 Chemical Modifications of Carotol and Their Antioxidant Activity

Authors: Dalvir Kataria, Khushminder Kaur Chahal, Amit Kumar

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The carrot seed essential oil was obtained by hydrodistillation. Hexane, dichloromethane, and methanol solvents were used for extraction of carrot seed by Soxhlet extraction methods. The major and minor compounds identified in carrot seed essential oil were carotol (52.73), daucol (5.10), daucene (5.68), (E)-β-farnesene (5.40), β-cubebene (3.19), longifolenaldehyde (3.23), β-elimene (3.23), (E)-caryophyllene (1.22), β-bisabolene (2.95) etc. The chemical composition of hexane, dichloromethane, and methanol extracts was different. Carotol was the common compound present. Major compounds isolated were from the carrot seed essential oil by column chromatography. Chemical transformations of carotol (2) with mercuric acetate/sodium borohydride, dry hydrochloric acid gas, acetonitrile/sulfuric acid, selenium dioxide/t-butyl hydrogen peroxide, N-bromosuccinimide, hydrogen iodide, and phenol were carried out. The derivatives of carotol were designed to explore the significance of some structural modifications in relation to antioxidant activities. The structures of major compounds and derivatives were confirmed on the basis of FT-IR, 1HNMR and 13CNMR spectroscopy. Antioxidant activity of carrot seed essential oil, various extracts and isolated compounds were tested by in vitro models involving 2, 2-diphenyl-1-picrylhydrazyl (DPPH•), hydroxyl (OH•), nitric oxide (NO•), superoxide radical scavenging methods and ferric reducing antioxidant power assay (FRAP). Chemical transformations of major isolated compound carotol were carried out, and antioxidant activity of all compounds was undertaken. The major sesquiterpenoidcarotol isolated from carrot seed essential oil showed the highest antioxidant activity in all the methods. The methanol extract showed higher antioxidant potential as compared to carrot seed essential oil, hexane, and dichloromethane extracts.

Keywords: antioxidant, carotol, carrot, DPPH

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716 Potential Application of Selected Halotolerant PSB Isolated from Rhizospheric Soil of Chenopodium quinoa in Plant Growth Promotion

Authors: Ismail Mahdi, Nidal Fahsi, Mohamed Hafidi, Abdelmounaim Allaoui, Latefa Biskri

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To meet the worldwide demand for food, smart management of arable lands is needed. This could be achieved through sustainable approaches such as the use of plant growth-promoting microorganisms including bacteria. Phosphate (P) solubilization is one of the major mechanisms of plant growth promotion by associated bacteria. In the present study, we isolated and screened 14 strains from the rhizosphere of Chenopodium quinoa wild grown in the experimental farm of UM6P and assessed their plant growth promoting properties. Next, they were identified by using 16S rRNA and Cpn60 genes sequencing as Bacillus, Pseudomonas and Enterobacter. These strains showed dispersed capacities to solubilize P (up to 346 mg L−1) following five days of incubation in NBRIP broth. We also assessed their abilities for indole acetic acid (IAA) production (up to 795,3 µg ml−1) and in vitro salt tolerance. Three Bacillus strains QA1, QA2, and S8 tolerated high salt stress induced by NaCl with a maximum tolerable concentration of 8%. Three performant isolates, QA1, S6 and QF11, were further selected for seed germination assay because of their pronounced abilities in terms of P solubilization, IAA production and salt tolerance. The early plant growth potential of tested strains showed that inoculated quinoa seeds displayed greater germination rate and higher seedlings growth under bacterial treatments. The positive effect on seed germination traits strongly suggests that the tested strains are growth promoting, halotolerant and P solubilizing bacteria which could be exploited as biofertilizers.

Keywords: phosphate solubilizing bacteria, IAA, Seed germination, salt tolerance, quinoa

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715 Prevalence of Mycoplasma hominis and Ureaplasma urealyticum as Causative Agents of Non-Gonococcal Urethritis in Men and Determination of Anti-Bacterial Resistance Rates

Authors: Recep Keşli, Cengiz Demir, Onur Türkyılmaz

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Objective: The aim of this study was to determine the prevalence of Mycoplasma hominis and Ureaplasma urealyticum as the causative agents in men with non-gonococcal urethtritis, and anti-bacterial resistance rates. Methods: The Study was carried out in the two Medical Microbiology Laboratories belonging to: Konya Education and Research Hospital and ANS Practice and Research Hospital, Afyon Kocatepe University, between January 2012 and December 2015. Urethral samples were obtained from patients by using a swab. Mycoplasma hominis and Ureaplasma urealyticum were detected by using Mycoplasma IST-2 kit (bio-Mérieux, Marcy l'Étoile, France). Neisseria gonorrhoea was excluded by Gram staining and culture methods. Results: Of all the one hundred and eighty-eight male patients with urethritis, forty M. hominis and forty two U. urealyticum were detected. Resistance rates of M. hominis strains against to doxycycline, ofloxacin, erythromycin, tetracycline, ciprofloxacin, azithromycin, clarithromycin, and pristinamycin were found as 5 %, 65 %, 25 %, 5 %, 80 %, 20 %, 20 %, 20 %, 5 %, respectively. Resistance rates of U. urealyticum strains against to doxycycline, ofloxacin, erythromycin, tetracycline, ciprofloxacin, azithromycin, clarithromycin, and pristinamycin were found as 4.7 %, 66.6 %, 23.8 %, 4.75 %, 81 %, 19 %, 19 %, 4.7 % respectively. No resistance was detected against to josamycin, for both the strains. Conclusions: It was concluded that; ciprofloxacin and ofloxacin had the weakest; josamycin, doxycycline, and tetracycline had the strongest in vitro anti-bacterial activity, for treatment of the NGU. So josamycin, doxycycline, and tetracycline should be preferred as the first choice of anti-bacterial agents, for treatment of the patients with non-gonococcal male urethritis.

Keywords: antimicrobial resistance, Mycoplasma hominis, non-gonococcal urethritis, Ureaplasma urealyticum

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714 Pragmatic Interpretation in Translated Texts

Authors: Jamal Alqinai

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A pragmatic approach to translation studies the rules and principles governing the use of language over and above the rules of syntax or morphology, and what makes some uses of language more appropriate than others in [communicative] situations. It attempts to explain translation as a procedure and product from the point of view of how, why and what is done by the source text author (ST) and what is to be done in the target text (TT) rendition. The latter will be subject to evaluation not as generated by the linguistics system but as conveyed and manipulated by participants in a communicative situation according to the referential and pragmatic standards employed. The failure of a purely lexical or structural translation stems from ignoring the relation between words as signs and the effect they have on their users. A more refined approach would also consider those processes that are sometimes labeled extra-linguistic or intuitive and which translators strive to reproduce unscathed in the translation process. We need to grasp the kind of actions an ST author performs on his readers by combining linguistic and non-linguistic elements against a backdrop of beliefs and cultural values. In other words, aside from considering the cohesive ties at the textual level, one needs to understand how the whole ST discourse hangs together logically in order to reproduce a coherent TT. The latter can only be achieved by an analysis of the pragmatic elements of presuppositions, implicatures and acts performed in the ST. Establishing cohesive ties within a text may require seeking reference outside the immediate text. The illocutionary functions manifested in one language/culture are relatively autonomous cultural/linguistic categories, but are imaginable by members of other cultures and, to some extent , are translatable though not, of course, without translation loss. Globalization and the spread of literacy worldwide may have created a universal empathy to comprehend the performative aspect of utterances when explained by approximate glosses or by paraphrase. Yet, it is often the multilayered and the culture-specific nature of illocutionary functions that de-universalize their possible interpretations. This paper addresses the pragmatic interpretation of culturally specific texts with examples adduced from a number of distinct settings to illustrate the influence of the pragmatic factors at stake.

Keywords: pragmatic, presupposition, implicature, cohesion

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713 Development of 3D Printed, Conductive, Biodegradable Nerve Conduits for Neural Regeneration

Authors: Wei-Chia Huang, Jane Wang

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Damage to nerves is considered one of the most irreversible injuries. The regeneration of nerves has always been an important topic in regenerative medicine. In general, damage to human tissue will naturally repair overtime. However, when the nerves are damaged, healed flesh wound cannot guarantee full restoration to its original function, as truncated nerves are often irreversible. Therefore, the development of treatment methods to successfully guide and accelerate the regeneration of nerves has been highly sought after. In order to induce nerve tissue growth, nerve conduits are commonly used to help reconnect broken nerve bundles to provide protection to the location of the fracture while guiding the growth of the nerve bundles. To prevent the protected tissue from becoming necrotic and to ensure the growth rate, the conduits used are often modified with microstructures or blended with neuron growth factors that may facilitate nerve regeneration. Electrical stimulation is another attempted treatment for medical rehabilitation. With appropriate range of voltages and stimulation frequencies, it has been demonstrated to promote cell proliferation and migration. Biodegradability are critical for medical devices like nerve conduits, while conductive polymers pose great potential toward the differentiation and growth of nerve cells. In this work, biodegradability and conductivity were combined into a novel biodegradable, photocurable, conductive polymer composite materials by embedding conductive nanoparticles in poly(glycerol sebacate) acrylate (PGSA) and 3D-printed into nerve conduits. Rat pheochromocytoma cells and rat neuronal Schwann cells were chosen for the in vitro tests of the conduits and had demonstrate selective growth upon culture in the conductive conduits with built-in microchannels and electrical stimulation.

Keywords: biodegradable polymer, 3d printing, neural regeneration, electrical stimulation

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712 Bronchoscopy and Genexpert in the Diagnosis of Pulmonary Tuberculosis in the Indian Private Health Sector: A Short Case Series

Authors: J. J. Mathew

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Pulmonary tuberculosis is highly prevalent in the Indian subcontinent. Most cases of pulmonary tuberculosis are diagnosed with sputum examinations and the vast majority of these are undertaken by the government run establishments. However, mycobacterial cultures are not routinely done, unless drug resistance is detected based on clinical response. Modern diagnostic tests like bronchoscopy and Genexpert are not routinely employed in the government institutions for the diagnosis of pulmonary tuberculosis, but have been accepted widely by good private institutions. The utility of these investigations in the private sector is not yet well recognized. This retrospective study aims to assess the usefulness of bronchoscopy and Genexpert in the diagnosis of pulmonary tuberculosis in quaternary care private hospital in India. 30 patients with respiratory symptoms raising the possibility of tuberculosis based on clinical and radiological features, but without any significant sputum production, were subject to bronchoscopy and BAL samples taken for microbiological studies, including Genexpert. 6 out of the 30 patients were found to be Genexpert positive and none of them showed Rifampicin resistance. All the 6 cases had upper zone predominant disease. One of the 6 cases of tuberculosis had another co-existent bacterial infection according to the routine culture studies. 6 other cases were proven to be due to other bacterial infections alone, 2 had a malignant diagnosis and the remaining cases were thought to be non-infective pathologies. The Genexpert results were made available within 48 hours in the 6 positive cases. All of them were commenced on standard anti-tuberculous regimen with excellent clinical response. The other infective cases were also managed successfully based on the drug susceptibilities. The study has shown the usefulness of these investigations as early intervention enabled diagnosis facilitating treatment and prevention of any clinical deterioration. The study lends support to early bronchoscopy and Genexpert testing in suspected cases of pulmonary tuberculosis without significant sputum production, in a high prevalence country which normally relies on sputum examination for the diagnosis of pulmonary tuberculosis.

Keywords: pulmonary, tuberculosis, bronchoscopy, genexpert

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711 An Analysis of the Representation of the Translator and Translation Process into Brazilian Social Networking Groups

Authors: Érica Lima

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In the digital era, in which we have an avalanche of information, it is not new that the Internet has brought new modes of communication and knowledge access. Characterized by the multiplicity of discourses, opinions, beliefs and cultures, the web is a space of political-ideological dimensions where people (who often do not know each other) interact and create representations, deconstruct stereotypes, and redefine identities. Currently, the translator needs to be able to deal with digital spaces ranging from specific software to social media, which inevitably impact on his professional life. One of the most impactful ways of being seen in cyberspace is the participation in social networking groups. In addition to its ability to disseminate information among participants, social networking groups allow a significant personal and social exposure. Such exposure is due to the visibility of each participant achieved not only on its personal profile page, but also in each comment or post the person makes in the groups. The objective of this paper is to study the representations of translators and translation process on the Internet, more specifically in publications in two Brazilian groups of great influence on the Facebook: "Translators/Interpreters" and "Translators, Interpreters and Curious". These chosen groups represent the changes the network has brought to the profession, including the way translators are seen and see themselves. The analyzed posts allowed a reading of what common sense seems to think about the translator as opposed to what the translators seem to think about themselves as a professional class. The results of the analysis lead to the conclusion that these two positions are antagonistic and sometimes represent conflict of interests: on the one hand, the society in general consider the translator’s work something easy, therefore it is not necessary to be well remunerated; on the other hand, the translators who know how complex a translation process is and how much it takes to be a good professional. The results also reveal that social networking sites such as Facebook provide more visibility, but it takes a more active role from the translator to achieve a greater appreciation of the profession and more recognition of the role of the translator, especially in face of increasingly development of automatic translation programs.

Keywords: Facebook, social representation, translation, translator

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710 Chromosomal Damage in Human Lymphocytes by Ultraviolet Radiation

Authors: Felipe Osorio Ospina, Maria Adelaida Mejia Arango, Esteban Onésimo Vallejo Agudelo, Victoria Lucía Dávila Osorio, Natalia Vargas Grisales, Lina María Martínez Sanchez, Camilo Andrés Agudelo Vélez, Ángela Maria Londoño García, Isabel Cristina Ortiz Trujillo

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Excessive exposure to ultraviolet radiation, has shown to be a risk factor for photodamage, alteration of the immune mechanisms to recognize malignant cells and cutaneous pro-inflamatorios States and skin cancers. Objective: Identify the time of exposure to ultraviolet radiation for the production of chromosomal damage in human lymphocytes. Methodology: We conducted an in vitro study serial, in which samples were taken from heparinized blood of healthy people, who do not submit exposure to agents that could induce chromosomal alterations. The samples were cultured in RPMI-1640 medium containing 10% fetal bovine serum, penicillin and streptomycin antibiotic. Subsequently, they were grouped and exposed to ultraviolet light for 1 to 20 seconds. At the end of the treatments, cytology samples were prepared, and it was colored with Giemsa (5%). Reading was carried out in an optical microscope and 100 metaphases analysed by treatment for posting chromosomal alterations. Each treatment was conducted at three separate times and each became two replicas. Results: We only presented chromosomal alterations in lymphocytes exposed to UV for a groups 1 to 3 seconds (p<0.05). Conclusions: Exposure to ultraviolet radiation generates visible damage in chromosomes from human lymphocytes observed in light microscopy, the highest rates of injury was observed between two and three seconds, and above this value, the reduction in the number of mitotic cells was evident.

Keywords: ultraviolet rays, lymphocytes, chromosome breakpoints, photodamage

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709 Ultraviolet Radiation and Chromosomal Damage in Human Lymphocytes

Authors: Felipe Osorio Ospina, Maria Adelaida Mejia Arango, Esteban Onésimo Vallejo Agudelo, Victoria Lucía Dávila Osorio, Natalia Vargas Grisales, Lina María Martínez Sanchez, Camilo Andrés Agudelo Vélez, Ángela Maria Londoño García, Isabel Cristina Ortiz Trujillo

Abstract:

Excessive exposure to ultraviolet radiation, has shown to be a risk factor for photodamage, alteration of the immune mechanisms to recognize malignant cells and cutaneous pro-inflamatorios states and skin cancers. Objective: To identify the time of exposure to ultraviolet radiation for the production of chromosomal damage in human lymphocytes. Methodology: We conducted an in vitro study serial, in which samples were taken from the heparinized blood of healthy people, who do not submit exposure to agents that could induce chromosomal alterations. The samples were cultured in RPMI-1640 medium containing 10% fetal bovine serum, penicillin, and streptomycin antibiotic. Subsequently, they were grouped and exposed to ultraviolet light for 1 to 20 seconds. At the end of the treatments, cytology samples were prepared, and it was colored with Giemsa (5%). Reading was carried out in an optical microscope and 100 metaphases analysed by treatment for posting chromosomal alterations. Each treatment was conducted at three separate times and each became two replicas. Results: We only presented chromosomal alterations in lymphocytes exposed to UV for groups 1 to 3 seconds (p < 0.05). Conclusions: Exposure to ultraviolet radiation generates visible damage in chromosomes from human lymphocytes observed in light microscopy, the highest rates of injury was observed between two and three seconds, and above this value, the reduction in the number of mitotic cells was evident.

Keywords: chromosome breakpoints, lymphocytes, photodamage, ultraviolet rays

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708 Development and Pre-clinical Evaluation of New ⁶⁴Cu-NOTA-Folate Conjugates for PET Imaging of Folate Receptor-Positive Tumors

Authors: Norah Al Hokbany, Ibrahim Al Jammaz, Basem Al Otaibi, Yousif Al Malki, Subhani M. Okarvi

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Objective: The folate receptor is over-expressed in a wide variety of human tumors. Conjugates of folate have been shown to be selectively taken up by tumor cells via the folate receptor. In an attempt to develop new folate radiotracers with favorable biochemical properties for detecting folate receptor-positive cancers. Methods: we synthesized ⁶⁴Cu-NOTA- and ⁶⁴Cu-NOTAM-folate conjugates using a straightforward and simple one-step reaction. Radiochemical yields were greater than 95% (decay-corrected) with a total synthesis time of less than 20 min. Results: Radiochemical purities were always greater than 98% without high-performance liquid chromatography (HPLC) purification. These synthetic approaches hold considerable promise as a rapid and simple method for ⁶⁴Cu-folate conjugate preparation with high radiochemical yield in a short synthesis time. In vitro tests on the KB cell line showed that significant amounts of the radio conjugates were associated with cell fractions. Bio-distribution studies in nude mice bearing human KB xenografts demonstrated a significant tumor uptake and favorable bio-distribution profile for ⁶⁴Cu-NOTA- and ⁶⁴Cu-NOTAM-folate conjugate. The uptake in the tumors was blocked by the excess injection of folic acid, suggesting a receptor-mediated process. Conclusion: These results demonstrate that the ⁶⁴Cu-NOTAM-folate conjugate may be useful as a molecular probe for the detection and staging of folate receptor-positive cancers, such as ovarian cancer and their metastasis, as well as monitoring tumor response to treatment.

Keywords: folate, receptor, tumor imaging, ⁶⁴Cu-NOTA-folate, PET

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707 The Effect of Technology on International Marketing Trading Researches and Analysis

Authors: Karim Monir Halim Salib

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The article discusses the use of modern technology to achieve environmental marketing goals in business and customer relations. The purpose of this article is to show the possibilities of the application of modern technology. In B2C relationships, marketing departments face challenges arising from the need to quickly segment customers and share information across multiple systems, which seriously hinders the achievement of marketing objectives. Therefore, the Article states that modern IT solutions are used in the marketing of business activities, taking into account environmental objectives. For this reason, its importance in the economic and social development of developing countries has increased. While traditional companies emphasize profit as the most important business principle, social enterprises have to address social issues at the expense of profit. This mindset gives social enterprises more than traditional businesses to meet the needs of those at the bottom of the pyramid. This also poses a great challenge for social business, as social business works for the public good on the one hand and financial stability on the other. Otherwise, the company cannot be evacuated. Cultures are involved in business communication and research. Using the example of language in international relations, the article poses the problem of cultural discourse in management and linguistic and cultural studies. After reviewing current research on language in international relations, this article presents communication methods in the international economy from a linguistic perspective and attempts to explain communication problems in business from the perspective of linguistic research. A step towards multidisciplinary research combining research in management and linguistics.

Keywords: international marketing, marketing mix, marketing research, small and medium-sized enterprises, strategic marketing, B2B digital marketing strategy, digital marketing, digital marketing maturity model, SWOT analysis consumer behavior, experience, experience marketing, marketing employee organizational performance, internal marketing, internal customer, direct marketing, mobile phones mobile marketing, Sms advertising.

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706 Formulation and Ex Vivo Evaluation of Solid Lipid Nanoparticles Based Hydrogel for Intranasal Drug Delivery

Authors: Pramod Jagtap, Kisan Jadhav, Neha Dand

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Risperidone (RISP) is an antipsychotic agent and has low water solubility and nontargeted delivery results in numerous side effects. Hence, an attempt was made to develop SLNs hydrogel for intranasal delivery of RISP to achieve maximum bioavailability and reduction of side effects. RISP loaded SLNs composed of 1.65% (w/v) lipid mass were produced by high shear homogenization (HSH) coupled ultrasound (US) method using glyceryl monostearate (GMS) or Imwitor 900K (solid lipid). The particles were loaded with 0.2% (w/v) of the RISP & surface-tailored with a 2.02% (w/v) non-ionic surfactant Tween® 80. Optimization was done using 32 factorial design using Design Expert® software. The prepared SLNs dispersion incorporated into Polycarbophil AA1 hydrogel (0.5% w/v). The final gel formulation was evaluated for entrapment efficiency, particle size, rheological properties, X ray diffraction, in vitro diffusion, ex vivo permeation using sheep nasal mucosa and histopathological studies for nasocilliary toxicity. The entrapment efficiency of optimized SLNs was found to be 76 ± 2 %, polydispersity index <0.3., particle size 278 ± 5 nm. This optimized batch was incorporated into hydrogel. The pH was found to be 6.4 ± 0.14. The rheological behaviour of hydrogel formulation revealed no thixotropic behaviour. In histopathology study, there was no nasocilliary toxicity observed in nasal mucosa after ex vivo permeation. X-ray diffraction data shows drug was in amorphous form. Ex vivo permeation study shows controlled release profile of drug.

Keywords: ex vivo, particle size, risperidone, solid lipid nanoparticles

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705 Brilliant Candy Consists of Centella asiatica Extract and Soy Milk to Safe Nutrition Child of Indonesia

Authors: Hesti Ghassani, Tessa Septiadi

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In the world we live on today, young generation highly influences the future of a nation. We have to concern that the condition of the country in 20 years later depending by the character of young adults these days. Therefore, it is important that we have to support and control the teenagers especially in one of developing countries in which I live in: Indonesia. Indonesia is a home to 240 million people. It diverse in languages, cultures, as well as attitudes. The differences among each individual lead us to think that there is something we have to take care of. It is necessary to pay attention to the nutrition consumed by the nation. We initiate to control the food consumed by young generation as early as a primary students. Nutrition affects the immune of the body, neuron system, and, most importantly brain. One of the nutrition that has to be fulfilled is milk. However, most of the population in Indonesia isn’t aware of the importance of consuming milk as their daily basis. We’ve formed an innovation called the Brilliant Candy which is affordable and rich in nutrition. So that is why the paper made by literature study to solve the problem with effective ways using available resources, practice and cheap. Brilliant Candy consists of Centella asiatica extract mixed with Soy milk. Centella asiatica contains of alkaloid which give the energy to brain and circulate oxygen. Based on the research of Sathya and Ganga, Centella asiatica can increase the intelligence. Indeed, Centella asiatica can relieve stress, and help us in staying focus. Soy milk is a kind of milk which come from extracted soybean. Soybean is rich in flafonoid. It has various advantages for our body. Which can also support child nutrition consumed. Soybean boosts immune system, helps digestive system, and in terms of food, soy bean exists as a source of nutrition. A method to get extraction of Centella asiatica is namely maserasi using ethanol. While making soybean milk with got the pollen of soybean. Both materials get mixed processed into hard candy with congelation of.

Keywords: Indonesia, Centella asiatica, Soy milk, alkaloid, flafonoid

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704 Lessons Learnt from a Patient with Pseudohyperkalaemia Secondary to Polycythaemia Rubra Vera in a Neuro-ICU Patient Resulting in Dangerous Interventions: Lessons Learnt on Patient Safety Improvement

Authors: Dinoo Kirthinanda, Sujani Wijeratne

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Pseudohyperkalaemia is a common benign in vitro phenomenon caused by the release of potassium ions (K+) from cells during specimen processing. Analysis of haemolysed blood samples for predominantly intracellular electrolytes may lead to re-investigation and potentially harmful interventions. We report a case of a 52-year male with myeloproliferative disease manifested as Polycythaemia Rubra Vera, Hypertension and hypertensive nephropathy with stage 3 chronic kidney disease admitted to Neuro-intensive care unit (NICU) with an intra-cerebral haemorrhage secondary to hypertensive bleed. His initial blood investigations showed hyperkalemia with serum K+ 6.2 mmol/L yet the bedside arterial blood gas analysis yielded K+ of 4.6 mmol/L. The patient was however given hyperkalemia regime twice based on venous electrolyte analysis. The discrepancy between the bedside electrolyte analysis using arterial blood and venous blood prompted further evaluation. The 12 lead Electrocardiogram showed U waves and sinus bradycardia corresponding to the serum K+ of 2.8 mmol/L on arterial blood gas analysis. Immediate K+ replacement ensured the patient did not develop life-threatening cardiac complications. Pseudohyperkalaemia may pose diagnostic challenges in the absence of detectable haemolysis and should be suspected in susceptible patients with normal Electrocardiogram and Glomerular Filtration Rate to avoid potentially life-threatening interventions. When in doubt, rapid analysis of arterial blood gas may be useful for accurate quantification of potassium.

Keywords: patient safety, pseudohyperkalaemia, haemolysis, myeloproliferative disorder

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703 Dilemma between the Education-Area and the Working-Area in Socialization of Teaching Profession: Scrutiny on the Beginning Teachers through the Relationality of the Regulations and Institutions in Turkey Case

Authors: Dilek Dede

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This study aims at scrutinized the dilemma between education place and working place with professional socialization dimension over the beginning teachers in Turkey is to be found the solution for the dilemma in Turkey. The research question is that how can be explained the gap between education place and working place for beginning teachers in Turkey. That expected to contribute to literature with the solutions for shorting the gap between working area and education area of the teaching profession in Turkey case. The study is constructed in two section. Firstly, socialization of the teaching profession and teaching modules have been discussed through the profession, education, working place indicators. In the second section, Secondly, two educational specialists from Turkey has been interviewed about their observation on trainee teachers compelling to participate the class for candidate teachers after university grade. Then, the dilemma between education area and working area of the teaching profession has been detected by of semi-structured and in-depth interviews, the literature on the relationality of institutions and regulation is discussed. The following outcomes have been accessed in accordance with the data set and literature linkage axis: Firstly, teachers coming from the distinctive programmes as an educational background. Hence, teachers who pertain to distinctive cultures work in the same environment. That cause cultural conflicts and complication of socialization of profession. Secondly, the insufficient partnership between schools and universities besides, the education classes lead to a struggle of culture among these two institutions. Thirdly, the education classes are designed as bureaucratic form instead of coalescence between head teachers and trainee teachers around a common culture. That become deep the dilemma. In conclusion, on condition that applied-oriented education that advocates in-service learning is promoted and this programme is supported with well-structured the in-service training through the partnership of universities and schools, the gap between the working-area and education-area might be shortened.

Keywords: beginning teachers, construction of a common, social mobilization in the teaching profession, teacher training institution, the relationality of the regulations and institutions

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702 Chrysin-Loaded PLGA-PEG Nanoparticles Designed for Enhanced Inhibitory Effect on the Breast Cancer Cell Line

Authors: Faraz Zarghami, Elham Anari, Nosratollah Zarghami, Yones Pilehvar-Soltanahmadi, Abolfazl Akbarzadeh, Sepideh Jalilzadeh-Tabrizi

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The development of nanotherapy has presented a new method of drug delivery targeted directly to the neoplasmic tissues, to maximize the action with fewer dose requirements. In the past two decades, poly(lactic-co-glycolic acid) (PLGA) has frequently been investigated by many researchers and is a popular polymeric candidate, due to its biocompatibility and biodegradability, exhibition of a wide range of erosion times, tunable mechanical properties, and most notably, because it is a FDA-approved polymer. Chrysin is a natural flavonoid which has been reported to have some significant biological effects on the processes of chemical defense, nitrogen fixation, inflammation, and oxidation. However, the low solubility in water decreases its bioavailability and consequently disrupts the biomedical benefits. Being loaded with PLGA-PEG increases chrysin solubility and drug tolerance, and decreases the discordant effects of the drug. The well-structured chrysin efficiently accumulates in the breast cancer cell line (T47D). In the present study, the structure and chrysin loading were delineated using proton nuclear magnetic resonance (HNMR), Fourier-transform infrared spectroscopy (FT-IR), and scanning electron microscopy (SEM), and the in vitro cytotoxicity of pure and nanochrysin was studied by the MTT assay. Next, the RNA was exploited and the cytotoxic effects of chrysin were studied by real-time PCR. In conclusion, the nanochrysin therapy developed is a novel method that could increase cytotoxicity to cancer cells without damaging the normal cells, and would be promising in breast cancer therapy.

Keywords: MTT assay, chrysin, flavonoids, nanotherapy

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