Search results for: mosquito cells

Authors: Ryan D. Martinus

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Inflammation of the endothelium is an important process leading to diabetic atherosclerosis. However, the molecular mechanisms by which diabetes contributes to endothelial inflammation remain to be established. Using In-vitro cultured Human cells and Hsp60 specific ELISA assays, we show that Hsp60 is not only induced in Human monocyte cells under hyperglycaemic conditions but that the Hsp60 is also secreted from these cells. Furthermore, we also demonstrate that the Hsp60 secreted from these monocyte cells is also able to activate Toll-like receptor-4 (TLR4) from Human endothelial cells. This suggests that a potential link may exist between the hyperglycaemia-induced expression of Hsp60 in monocyte cells and vascular inflammation. Circulating levels of Hsp60 due to mitochondrial stress in diabetes patients could, therefore, be an important modulator of inflammation in endothelial cells and thus contribute to the increased incidences of atherosclerosis in diabetes mellitus.

Keywords: mitochondria, Hsp60, inflammation, diabetes mellitus

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Authors: Piamsiri Sawaisorn, Tienrat Tangchaikeeree, Waraporn Chan-On, Chaniya Leepiyasakulchai, Rachanee Udomsangpetch, Suradej Hongeng, Kulachart Jangpatarapongsa

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Human Vγ9Vδ2 T lymphocytes are regarded as promising effector cells for cancer immunotherapy since they have the ability to eliminate several tumor cells through non-peptide antigen recognition and non-major histocompatibility complex (MHC) restriction. An issue of recent interest is the capability to activate γδ T cells by use of a group of drugs, such as pamidronate, that cause accumulation of phosphoantigen which is recognized by γδ T cell receptors. Moreover, their antigen presenting cell-like phenotype and function have been confirmed in many clinical trials. In this study, Vγ9Vδ2 T cells derived from normal peripheral blood mononuclear cells were activated with pamidronate and the expanded Vγ9Vδ2 T cells can recognize and kill chronic myeloid leukemia (CML) cells treated with pamidronate through their cytotoxic activity. To support the strong role played by Vγ9Vδ2 T cells against cancer, we provide the evidence that Vγ9Vδ2 T cells activated with CML cell lysate antigen can efficiently express antigen presenting cell (APC) phenotype and function. In conclusion, pamidronate can be used in intentional activation of human Vγ9Vδ2 T cells and can increase the susceptibility of CML cells to cytotoxicity of Vγ9Vδ2 T cells. The activated Vγ9Vδ2 T cells by cancer cells lysate can show their APC characteristics, and so greatly increase the interest in exploring their therapeutic potential in hematologic malignancy.

Keywords: γδ T lymphocytes, antigen-presenting cells, chronic myeloid leukemia, cancer, immunotherapy

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Authors: Fatma Y. Meligy

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Objectives: The objectives of this study aimed to isolate and characterize mesenchymal stem cells (MSCs) derived from synovial membrane. Then to assess the potentiality of myogenic differentiation of these isolated MSCs. Methods: The MSCs were isolated from synovial membrane by digestion method. Three adult rats were used. The 5 -azacytidine was added to the cultured cells for one day. The isolated cells and treated cells are assessed using immunoflouresence, flowcytometry, PCR and real time PCR. Results: The isolated stem cells showed morphological aspect of stem cells they showed strong positivity to CD44 and CD90 in immunoflouresence while in CD34 and CD45 showed negative reaction. The treated cells with 5-azacytidine was shown to have positive reaction for desmin. Flowcytometric analysis showed that synovial MSCs had strong positive percentage for CD44(%98)and CD90 (%97) and low percentage for CD34 & CD45 while the treated cells showed positive percentage for myogenic marker myogenin (85%). As regard the PCR and Real time PCR, the treated cells showed positive reaction to the desmin primer. Conclusion: The adult MSCs were isolated successfully from synovial membrane and characterized with stem cell markers. The isolated cells could be differentiated in vitro into myogenic cells. These differentiated cells could be used in auto-replacement of diseased or traumatized muscle cells as a regenerative therapy for muscle disorders and trauma.

Keywords: mesenchymal stem cells, synovial membrane, myogenic differentiation

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Authors: Byeal-I Han, Michael Lee

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Ageing is associated with an increased risk of diseases such as cancer, and neurodegenerative disorders. Increased autophagy delays ageing and extends longevity. In this study, we investigated the role of autophagy in longevity using human foreskin fibroblast HS68 cells, in which a senescence-like growth arrest can be induced. In particular, cellular senescence is manifested by the irreversible cell cycle arrest, and may contribute to the ageing of organisms. The senescence state was measured with staining for senescence-associated β-galactosidase (SA-β-gal) activity that represents a sensitive and reliable marker to quantify senescent cells. We detected a significantly increased percentage (%) of SA-β-gal positive cells in HS68 cultures at passage 40 (63%) when compared with younger ones at passage 15 (0.5%). As expected, HS68 cells at passage 40 exhibited much lower proliferation rate than cells at passage 15. The basal levels of LC3 were measured by immunoblotting showing a comparison of LC3-I and LC3-II levels at 3 age-points in serially passaged HS68 cells. LC3-II/LC3-I ratio at different passage levels relative to β-actin levels of each band confirmed that cells at passage 34 showed lower conversion of non-autophagic LC3-I to autophagic LC3-II than the cells at passage 16. Furthermore, Cyto-ID autophagy assay also revealed that late passage cells showed lower autophagy than the early passage cells. Together, our findings suggest that senescence induction might be associated with decreased autophagy.

Keywords: ageing, autophagy, senescence, HS68

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Authors: Sung-Hee Hwang, Michael Lee

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Upregulated Src activity has been implicated in a variety of cancers. Thus, Src family tyrosine kinase (SFK) inhibitors are often effective cancer treatments. Here, we investigate the role of autophagy in ATG5 knockout cell lines generated by the Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas mediated genome editing. The CRISPR-associated protein Cas9 is an RNA-guided DNA endonuclease that uses RNA–DNA complementarity to identify target sites for sequence specific double-stranded DNA (dsDNA) cleavage. Interestingly, ATG5 KO cells clearly showed a greater proliferation rate than WT NIH 3T3 cells, implying that autophagy induction is cytotoxic. Also, the clonogenic survival of ATG5 KO cells was greater than WT cells. The MTT assay revealed that the cytotoxic effect of PP2 was weaker on ATG5 knockout cells than that WT cells. The conversion of non-autophagic LC3-I to autophagic LC3-II and RT-PCR confirmed the functional gene knockout. Furthermore, Cyto-ID autophagy assay also revealed that PP2 failed to induce autophagy in ATG5 knockout cells. Together, our findings suggest that the resistance to PP2 in ATG5 knockout cells is associated with defective autophagy.

Keywords: ATG5 knockout, Autophagy, CRISPR/Cas9, PP2

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Authors: Rauza Sukma Rita, Katsuya Dezaki, Yuko Maejima, Toshihiko Yada

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Oxytocin is a nine-amino acid peptide synthesized in the paraventricular nucleus (PVN) and supraoptic nucleus (SON) of the hypothalamus. Oxytocin promotes contraction of the uterus during birth and milk ejection during breast feeding. Although oxytocin receptors are found predominantly in the breasts and uterus of females, many tissues and organs express oxytocin receptors, including the pituitary, heart, kidney, thymus, vascular endothelium, adipocytes, osteoblasts, adrenal gland, pancreatic islets, and many cell lines. On the other hand, in pancreatic islets, oxytocin receptors are expressed in both α-cells and β-cells with stronger expression in α- cells. However, to our knowledge there are no reports yet about the effect of oxytocin on cytosolic calcium reaction on α and β-cell. This study aims to investigate the effect of oxytocin on α-cells and β-cells and its oscillation pattern. Islet of Langerhans from wild type mice were isolated by collagenase digestion. Isolated and dissociated single cells either α-cells or β-cells on coverslips were mounted in an open chamber and superfused in HKRB. Cytosolic concentration ([Ca2+]i) in single cells were measured by fura-2 microfluorimetry. After measurement of [Ca2+]i, α-cells were identified by subsequent immunocytochemical staining using an anti-glucagon antiserum. In β-cells, the [Ca2+]i increase in response to oxytocin was observed only under 8.3 mM glucose condition, whereas in α-cells, [Ca2+]i an increase induced by oxytocin was observed in both 2.8 mM and 8.3 mM glucose. The oscillation incidence was induced more frequently in β-cells compared to α-cells. In conclusion, the present study demonstrated that oxytocin directly interacts with both α-cells and β-cells and induces increase of [Ca2+]i and its specific patterns.

Keywords: α-cells, β-cells, cytosolic calcium concentration, oscillation, oxytocin

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Authors: Bhaskar M. Murai, Neeraj Banchor, Ishveen Chabbra, Madhusudhan Nadgir, S. Vidhya

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Sol-gel technology combined with electronics and biochemistry helps to overcome the problems caused by mosquitoes by developing a portable, low-cost device which enables controlled release of trapped compound inside it. It is a wet-chemical technique which is used primarily for fabrication of silicate gel which is usually allowed to dry as per requirement. The outcome is solid rock hard material which is porous and has lots of applications in different fields. Taking porosity as a key factor, allethrin a naturally occurring synthetic compound with molecular mass 302.40 was entrapped inside the sol-gel matrix as a dopant. Allethrin is commonly used as an insecticide and is a key ingredient in commercially available mosquitoes repellent in Asian and subtropical countries. It has low toxicity for humans and birds, and are used in many household insecticides such as RAID as well as mosquito coils. They are however highly toxic to fish and bees. Insects subject to its exposure become paralyzed (nervous system effect) before dying. They are also used as an ultra-low volume spray for outdoor mosquito control. Therefore, there is a need for controlled release of allethrin in the environment. For controlled release of allethrin from sol-gel matrix, its (allethrin) we utilized temperature based controlled evaporation through porous sol-gel. Different types of fabric like cotton, Terri-cotton, polyester, surgical cap, knee-cap etc are studied and the best with maximum absorption capacity is selected to hold the sol-gel matrix with maximum quantity. For sol-gel coating 2 x 2cm cloth pieces are dipped in sol-gel solution for 10 minutes and by calculating the weight difference we concluded that Terri cotton is best suitable for our project. An electronic circuit with heating plate is developed in to test the controlled release of compound. An oscillatory circuit is used to produce the required heat.

Keywords: sol-gel, allethrin, TEOS, biochemistry

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Authors: Samia Moulebhar, Chahrazed Bendenia, Souhila Bendenia, Hanaa Merad-dib, Sarra Merabet, Sid Ahmed Khantar, Baghdad Hadri

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The photovoltaic solar field is today experiencing exponential advancement with the exploitation of new technological sectors of nanoparticles, namely the field of solar cells based on organic polymer materials. These cells are flexible, easy to process and low cost. This work includes a presentation of the conjugated polymer materials used in the design of photovoltaic technology devices while determining their properties and then the models used for the modeling of thin film photovoltaic cells heterojunction.

Keywords: photovoltaic, cells, nanoparticles, organic

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Authors: M. D. Nirmani, K. L. N. Perera, G. H. Galhena

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Dengue has become one of the most important arbo-viral disease in all tropical and subtropical regions of the world. Aedes aegypti, is the principal vector of the virus, vary in both epidemiological and behavioral characteristics, which could be finely measured through DNA sequence comparison at their population level. Such knowledge in the population differences can assist in implementation of effective vector control strategies allowing to make estimates of the gene flow and adaptive genomic changes, which are important predictors of the spread of Wolbachia infection or insecticide resistance. As such, this study was undertaken to investigate the phylogenetic relationships of Ae. aegypti from Galle and Colombo, Sri Lanka, based on the ribosomal protein region which spans between two exons, in order to understand the geographical distribution of genetically distinct mosquito clades and its impact on mosquito control measures. A 320bp DNA region spanning from 681-930 bp, corresponding to the ribosomal protein, was sequenced in 62 Ae. aegypti larvae collected from Galle (N=30) and Colombo (N=32), Sri Lanka. The sequences were aligned using ClustalW and the haplotypes were determined with DnaSP 5.10. Phylogenetic relationships among haplotypes were constructed using the maximum likelihood method under Tamura 3 parameter model in MEGA 7.0.14 including three previously reported sequences of Australian (N=2) and Brazilian (N=1) Ae. aegypti. The bootstrap support was calculated using 1000 replicates and the tree was rooted using Aedes notoscriptus (GenBank accession No. KJ194101). Among all sequences, nineteen different haplotypes were found among which five haplotypes were shared between 80% of mosquitoes in the two populations. Seven haplotypes were unique to each of the population. Phylogenetic tree revealed two basal clades and a single derived clade. All observed haplotypes of the two Ae. aegypti populations were distributed in all the three clades, indicating a lack of genetic differentiation between populations. The Brazilian Ae. aegypti haplotype and one of the Australian haplotypes were grouped together with the Sri Lankan basal haplotype in the same basal clade, whereas the other Australian haplotype was found in the derived clade. Phylogram showed that Galle and Colombo Ae. aegypti populations are highly related to each other despite the large geographic distance (129 Km) indicating a substantial genetic similarity between them. This may have probably arisen from passive migration assisted by human travelling and trade through both land and water as the two areas are bordered by the sea. In addition, studied Sri Lankan mosquito populations were closely related to Australian and Brazilian samples. Probably this might have caused by shipping industry between the three countries as all of them are fully or partially enclosed by sea. For example, illegal fishing boats migrating to Australia by sea is perhaps a good mean of transportation of all life stages of mosquitoes from Sri Lanka. These findings indicate that extensive mosquito migrations occur between populations not only within the country, but also among other countries in the world which might be a main barrier to the successful vector control measures.

Keywords: Aedes aegypti, dengue control, extensive mosquito migration, haplotypes, phylogeny, ribosomal protein

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Authors: Maria C. Proença, Maria T. Rebelo, Marília Antunes, Maria J. Alves, Hugo Osório, Sofia Cunha, João Casaca

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Using a geographic information system (GIS), the relations between a georeferenced data set of Culex pipiens sl. mosquitoes collected in Portugal mainland during seven years (2006-2012) and meteorological and physiographic parameters such as: air relative humidity, air temperature (minima, maxima and mean daily temperatures), daily total rainfall, altitude, land use/land cover and proximity to water bodies are evaluated. Focus is on the mosquito females; the characterization of its habitat is the key for the planning of chirurgical non-aggressive prophylactic countermeasures to avoid ambient degradation. The GIS allow for the spatial determination of the zones were the mosquito mean captures has been above average; using the meteorological values at these coordinates, the limits of each parameter are identified/computed. The meteorological parameters measured at the net of weather stations all over the country are averaged by month and interpolated to produce raster maps that can be segmented according to the thresholds obtained for each parameter. The intersection of the maps obtained for each month show the evolution of the area favorable to the species through the mosquito season, which is from May to October at these latitudes. In parallel, mean and above average captures were related to the physiographic parameters. Three levels of risk could be identified for each parameter, using above average captures as an index. The results were applied to the suitability meteorological maps of each month. The Culex pipiens critical niche is delimited, reflecting the critical areas and the level of risk for transmission of the pathogens to which they are competent vectors (West Nile virus, iridoviruses, rheoviruses and parvoviruses).

Keywords: Culex pipiens, ecological niche, risk assessment, risk management

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Authors: Merve Colak, Gizem Donmez Yalcin

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Glutamate has many neurological functions in the central nervous system and is found at high concentrations in the brain. Increased levels of glutamate in the neuronal space are toxic, causing neuron damage and death. This is called glutamate-induced excitotoxicity. Excitotoxicity is among the causes of many neurological diseases such as trauma, cerebral ischemia, epilepsy, Parkinson's Disease, Alzheimer's Disease. Since neuroblastoma cells are known to be excitotoxic, we propose that excitotoxicity can be studied in neuroblastoma cells. Excitotoxicity can be induced using kainic acid in neuroblastoma cells. Measuring the secretion of glutamate, excitotoxicity can be analyzed in neuroblastoma cells.

Keywords: glutamate, excitotoxicity, kainic acid, Sirt4

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Authors: Hirowati Ali, Aisyah Ellyanti, Dewi Rusnita, Septelia Inawati Wanandi

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Stem cell has been known for its potency to be differentiated in many cells. Recently stem cell has been used for many treatment of degenerative medicine. It is still controversy whether stem cell can be used for therapy or these cells can activate cancer stem cell. SCUBE2 is a novel secreted and membrane-anchored protein which has been reported to its role in better prognosis and inhibition of cancer cell proliferation. Our study aims to observe whether stem cell can up-regulate SCUBE2 gene in MCF7 breast cancer cell line. We used in vitro study using MCF-7 cell treated with stem cell derived from placenta Wharton's jelly which has been known for its stemness and widely used. Our results showed that MCF-7 cell line grows up rapidly in 6-well culture dish. Stem cell was cultured in 6-well dish. After 50%-60% MCF-7 confluence, we co-cultured these cells with stem cells for 24 hours and 48 hours. We hypothesize SCUBE2 gene which is previously known for its higher expression in better prognosis of breast cancer, is up-regulated after stem cells addition in MCF7 culture dishes.

Keywords: breast cancer cells, inhibition of cancer cells, mesenchymal stem cells, SCUBE2

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Authors: Chiu-Wen Chang, I-Yun Chang, Wei-Ting Chen, Hui-Ping Ho, Chao-Ying Pan, Joh-Jong Huang

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Dengue is a ‘community disease’ or ‘environmental disease’, as long as the environment exist suitable container (including natural and artificial) for mosquito breeding, once the virus invade will lead to the dengue epidemic. Surveillance of vector density is critical to effective infectious disease control and play an important role in monitoring the dynamics of mosquitoes in community, such as mosquito species, density, distribution area. The objective of this study was to examine the relationship in vector density survey (Breteau index, Adult index, House index, Container index, and Larvae index) form 2014 to 2016 in Kaohsiung City and evaluate the effects of introducing the Breeding Elimination and Appraisal Team (hereinafter referred to as BEAT) as an intervention measure on eliminating dengue vector breeding site started from May 2016. BEAT were performed on people who were suspected of contracting dengue fever, a surrounding area measuring 50 meters by 50 meters was demarcated as the emergency prevention and treatment zone. BEAT would perform weekly vector mosquito inspections and vector mosquito inspections in regions with a high Gravitrap index and assign a risk assessment index to each region. These indices as well as the prevention and treatment results were immediately reported to epidemic prevention-related units every week. The results indicated that, vector indices from 2014 to 2016 showed no statistically significant differences in the Breteau index, adult index, and house index (p > 0.05) but statistically significant differences in the container index and larvae index (p <0.05). After executing the integrated elimination work, container index and larvae index are statistically significant different from 2014 to 2016 in the (p < 0.05). A post hoc test indicated that the container index of 2014 (M = 12.793) was significantly higher than that of 2016 (M = 7.631), and that the larvae index of 2015 (M = 34.065) was significantly lower than that of 2014 (M = 66.867). The results revealed that effective vector density surveillance could highlight the focus breeding site and then implement the immediate control action (BEAT), which successfully decreased the vector density and the risk of dengue epidemic.

Keywords: Breteau index, dengue control, monitoring method, vector density

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Authors: Vandana Gawande, Chandani Satija

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Gnidia glauca is a semi-woody herb of thymelaeaceae family traditionally used as fish poison in India. It is also found in Sri lanka and Africa. In the present study, potential anticancer effect of n-hexane and ethanolic extracts of Gnidia glauca in human breast cancer cells was investigated. Human breast cancer cells (MCF-7) were cultured as monolayers in RPMI 1640 medium. The cells were cultured for 48 hours to allow growth and achieve about 80% confluence in 96-well culture plates. The cells were treated with various concentrations of Gnidia glauca (0.1-100 mg/mL) for 72 hours. Percentage of viable cells after treatment was assessed using a sulforhodamine B colorimetric assay. Both n-hexane and ethanolic extract showed significant cytotoxic activity on MCF-7 cancer cells. This study supports the notion of using Gnidia glauca as a novel anticancer agent for breast cancer.

Keywords: 96 well plate, anticancer activity, Gnidia glauca, MCF-7

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Authors: Aasaithambi Kalaiselvi, Michael Gabriel Paulraj, Ekambaram Nakkeeran

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Emergences of mosquito vector-borne diseases are considered as a perpetual problem globally in tropical countries. The outbreak of several diseases such as chikungunya, zika virus infection and dengue fever has created a massive threat towards the living population. Frequent usage of synthetic insecticides like Dichloro Diphenyl Trichloroethane (DDT) eventually had its adverse harmful effects on humans as well as the environment. Since there are no perennial vaccines, prevention, treatment or drugs available for these pathogenic vectors, WHO is more concerned in eradicating their breeding sites effectively without any side effects on humans and environment by approaching plant-derived natural eco-friendly bio-insecticides. The aim of this study is to investigate the larvicidal potency of Cinnamomum verum essential oil (CEO) loaded niosomes. Cholesterol and surfactant variants of Span 20, 60 and 80 were used in synthesizing CEO loaded niosomes using Transmembrane pH gradient method. The synthesized CEO loaded niosomes were characterized by Zeta potential, particle size, Fourier Transform Infrared Spectroscopy (FT-IR), GC-MS and SEM analysis to evaluate charge, size, functional properties, the composition of secondary metabolites and morphology. The Z-average size of the formed niosomes was 1870.84 nm and had good stability with zeta potential -85.3 meV. The entrapment efficiency of the CEO loaded niosomes was determined by UV-Visible Spectrophotometry. The bio-potency of CEO loaded niosomes was treated and assessed against gram-positive (Bacillus subtilis) and gram-negative (Escherichia coli) bacteria and fungi (Aspergillus fumigatus and Candida albicans) at various concentrations. The larvicidal activity was evaluated against II to IV instar larvae of Aedes aegypti, Anopheles stephensi and Culex quinquefasciatus at various concentrations for 24 h. The mortality rate of LC₅₀ and LC₉₀ values were calculated. The results exhibited that CEO loaded niosomes have greater efficiency against mosquito larvicidal activity. The results suggest that niosomes could be used in various applications of biotechnology and drug delivery systems with greater stability by altering the drug of interest.

Keywords: Cinnamomum verum, niosomes, entrapment efficiency, bactericidal and fungicidal, mosquito larvicidal activity

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Authors: Ga-Young Lee, Hyun-Man Kim

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The modulation of the cell-cell junction is critical in epithelial-mesenchymal transition during tumorigenesis. As RhoA activity is known to be up-regulated to dissociate cell-cell junction by contracting acto-myosin complex in various cancer cells, the present study investigated if RhoA activity was also associated with the disruption of the cell-cell junction of oral cancer cells. We studied SCC-25 cells which are established from oral squamous cell carcinoma if their E-cadherin junction (ECJ) was under control of RhoA. Interestingly, development of ECJ of SCC-25 cells depended on the amount of fibronectin (FN) coated on the culture dishes. Seeded cells promptly aggregated to develop ECJ on the substrates coated with a low amount of FN, whereas they were retarded in the development of ECJ on the substrates coated with a high amount of FN. However, it was an unexpected finding that total RhoA activity was lower in the dissociated cells on the substrates of high FN than in the aggregated cells on the substrates of low FN. Treating the dissociated cells on the substrates of high FN with LPA, a RhoA activator, promoted the development to ECJ. In contrast, treating the aggregated cells on the substrates of low FN with Clostridium botulinum C3, a toxin decreasing RhoA activity, dissociated cells concomitant with the disruption of ECJ. Genetical knockdown of RhoA expression by transfecting RhoA siRNA also down-regulated the development of ECJ in SCC-25 cells. Furthermore, PMA, an activator of protein kinase C (PKC), down-regulated the development of ECJ junction of SCC-25 cells on the substrates coated with low FN. In contrast, GO6976, a PKC inhibitor, up-regulated the development of ECJ of SCC-25 cells with the activation of RhoA on the substrates coated with high FN. In conclusion, in the present study, we demonstrated unexpected results that the activation of RhoA promotes the development of ECJ, whereas the inhibition of RhoA retards the development of ECJ in SCC-25 cells.

Keywords: E-cadherin junction, oral squamous cell carcinoma, PKC, RhoA, SCC-25

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Authors: Dipali Dhawan

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Cancerous epithelial cells are confined to a primary site by the continued expression of adhesion molecules and the intact basal lamina. However, as the cancer progresses some cells are believed to undergo an epithelial-mesenchymal transition (EMT) event, leading to increased motility, invasion and, ultimately, metastasis of the cells from the primary tumour to secondary sites within the body. These disseminated cancer cells need the ability to self-renew, as stem cells do, in order to establish and maintain a heterogeneous metastatic tumour mass. Identification of the specific subpopulation of cancer stem cells amenable to the process of metastasis is highly desirable. In this study, we have isolated and characterized cancer stem cells from luminal and basal breast cancer cell lines (MDA-MB-231, MDA-MB-453, MDA-MB-468, MCF7 and T47D) on the basis of cell surface markers CD44 and CD24; as well as Side Populations (SP) using Hoechst 33342 dye efflux. The isolated populations were analysed for epithelial and mesenchymal markers like E-cadherin, N-cadherin, Sfrp1 and Vimentin by Western blotting and Immunocytochemistry. MDA-MB-231 cell lines contain a major population of CD44+CD24- cells whereas MCF7, T47D and MDA-MB-231 cell lines show a side population. We observed higher expression of N-cadherin in MCF-7 SP cells as compared to MCF-7NSP (Non-side population) cells suggesting that the SP cells are mesenchymal like cells and hence express increased N-cadherin with stem cell-like properties. There was an expression of Sfrp1 in the MCF7- NSP cells as compared to no expression in MCF7-SP cells, which suggests that the Wnt pathway is expressed in the MCF7-SP cells. The mesenchymal marker Vimentin was expressed only in MDA-MB-231 cells. Hence, understanding the breast cancer heterogeneity would enable a better understanding of the disease progression and therapeutic targeting.

Keywords: cancer stem cells, epithelial to mesenchymal transition, biomarkers, breast cancer

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Authors: Laura Bellassen, Idit Shachar

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Multiple sclerosis (MS) is a chronic neurological disorder characterized by demyelination of the central nervous system (CNS), leading to a wide range of physical and cognitive impairments. Myeloid cells in the CNS, such microglia and border associated macrophage cells, participate in the neuroinflammation in MS. Activation of those cells in MS contributes to the inflammatory response in the CNS and recruitment of immune cells in the this compartment. SLAMF5 is a cell surface receptor that functions as a homophilic adhesion molecule, whose signaling can activate or inhibit leukocyte function. In the current study we followed the expression and function of SLAMF5 in myeloid cells in the CNS and in the periphery in the murine model for MS, the experimental autoimmune encephalomyelitis model (EAE). Our results show that SLAMF5 deficiency or blocking decreases the expression of activation molecules and costimulatory molecules such as MHCII and CD80, resulting in delayed onset and reduced progression of the disease. Moreover, blocking SLAMF5 in peripheral monocytes derived from MS patients and iPSC-derived microglia cells, controls the expression of HLA-DR and CD80. Thus, SLAMF5 is a regulator of myeloid cells function and can serve as a therapeutic target in autoimmune disorders as Multiple Sclerosis.

Keywords: multiple sclerosis, EAE model, myeloid cells, new antibody, neuroimmunology

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Authors: E. Marei, A. Hammad, S. Ismail, A. El-Gindy

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This study aims to investigate the ability of different formula of mixed bacteria as a biological treatments of wastewater after primary treatment as a bio-treatment and bio-removal and bio-adsorbent of different heavy metals in natural circumstances. The wastewater was collected from Sarpium forest site-Ismailia Governorate, Egypt. These treatments were mixture of free cells and mixture of immobilized cells of different bacteria. These different formulas of mixed bacteria were prepared under Lab. condition. The obtained data indicated that, as a result of wastewater bio-treatment, the removal rate was found to be 76.92 and 76.70% for biological oxygen demand, 79.78 and 71.07% for chemical oxygen demand, 32.45 and 36.84 % for ammonia nitrogen as well as 91.67 and 50.0% for phosphate after 24 and 28 hrs with mixed free cells and mixed immobilized cells, respectively. Moreover, the bio-removals of different heavy metals were found to reach 90.0 and 50. 0% for Cu ion, 98.0 and 98.5% for Fe ion, 97.0 and 99.3% for Mn ion, 90.0 and 90.0% Pb, 80.0% and 75.0% for Zn ion after 24 and 28 hrs with mixed free cells and mixed immobilized cells, respectively. The results indicated that 13.86 and 17.43% of removal efficiency and reduction of total dissolved solids were achieved after 24 and 28 hrs with mixed free cells and mixed immobilized cells, respectively.

Keywords: wastewater bio-treatment , bio-sorption heavy metals, biological desalination, immobilized bacteria, free cell bacteria

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Authors: Soumitra Satapathi, Anubhav Raghav

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Perovskite solar cell technology has passed through a phase of unprecedented growth in the efficiency scale from 3.8% to above 22% within a half decade. This technology has drawn tremendous research interest. It has been observed that performances of perovskite based solar cells are extremely dependent on the morphology and crystallinity of the perovskite layer. It has also been observed that device lifetime depends on the perovskite morphology; devices with larger perovskite grains degrade slowly than those of the smaller ones. Various methods of perovskite growth have been applied to achieve the most appropriate morphology necessary for high efficient solar cells. The recent progress in morphology optimization by various methods emphasizing on grain sizes, stoichiometry, and ambient compatibility as well as photophysics study in air-processed perovskite solar cells will be discussed.

Keywords: perovskite solar cells, morphology optimization, photophysics study, air-processed solar cells

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Authors: Surachart Koyadun

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Introduction: Plasmodium knowlesi (P. knowlesi) malaria is a zoonotic disease that is classified as type 5 of human malaria. Commonly found in macaques (Macaca fascicularis) and (Macaca nemestrina), P. knowlesi is capable of resulting in both uncomplicated and severe malaria in humans. Situation of P. knowlesi malaria in Phang-Nga province for the past 3 years from 2020 – 2022 revealed no case report in 2020, however, a total of 14 cases had been reported in 2021 - 2022. This research aimed to 1) study the epidemiology of P. knowlesi, 2) examine the clinical manifestations of P. knowlesi patients, 3) analyze the ecology and entomology of P. knowlesi, and 4) analyze the diagnosis and treatment of P. knowlesi. Method: This research was a retrospective descriptive study/case report. The study was conducted in 14 patients with P. knowlesi malaria between 2021 and 2022 in 4 districts of Phang-Nga Province, Thailand including Thapput, Kapong, Takuapa and Khuraburi. Results: The study subjects of P. knowlesi malaria were all males. Most of them were working age groups as farmers and worked in forest or plantation areas. All had no history of blood transfusions. Most of the patients did not use mosquito nets and had a history of camping in the forest prior to the onset of fever. An analysis of all 14 sources of infection unveiled the area is home to macaques, and that area has detected Anopheles mosquito, which is the carrier of the disease. Majority of them got sick in the dry season of Thailand (December-April). The main symptoms brought to the hospital were fever, chills, headache, body aches. Laboratory findings on the first day of diagnosis were as follows: The white blood cell count was found within the normal range. In the proportion of white blood cells, eosinophils were found to be slightly higher than normal. Slight anemia was found on early examination. The platelet count was found to be below normal in all cases. Severely low platelet count (2,000 cells/mm3) was found in severe cases with multiple complications. No patient was found dead but 85.7% of complications were found, with acute renal failure being the most common. Patients with delayed diagnosis and treatment of malaria (inaccurate diagnosis or late access to the hospital) had the highest severity and complications than those who had seen the doctor since the first 3-4 days of illness or the screening of symptoms and risk history by the malaria clinic staff at vector-borne disease control unit. Conclusion and Recommendation: P. knowlesi malaria is an emerging infectious disease transmitted from animals to humans. There are challenges in epidemiology, entomology, ecology for effective surveillance, prevention and control. Early diagnosis and treatment would reduce complications and prevent death.

Keywords: malaria, plasmodium knowlesi, epidemiology, ecology, entomology, diagnosis, treatment

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Authors: Aliya Sekenova, Vyacheslav Ogay

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The derivation of human induced pluripotent stem cells (iPSCs) from somatic cells by direct reprogramming opens wide perspectives in the regenerative medicine. It means the possibility to develop the personal and, consequently, any immunologically compatible cells for applications in cell-based therapy. The purpose of our study was to develop the technology for the production of NK cells from T cell-derived induced pluripotent stem cells (TiPSCs) for subsequent application in adoptive cancer immunotherapy. Methods: In this study iPSCs were derived from peripheral blood T cells using Sendai virus vectors expressing Oct4, Sox2, Klf4 and c-Myc. Pluripotent characteristics of TiPSCs were examined and confirmed with alkaline phosphatase staining, immunocytochemistry and RT-PCR analysis. For NK cell differentiation, embryoid bodies (EB) formed from (TiPSCs) were cultured in xenogeneic serum-free medium containing human serum, IL-3, IL-7, IL-15, SCF, FLT3L without using M210-B4 and AFT-024 stromal feeder cells. After differentiation, NK cells were characterized with immunofluorescence analysis, flow cytometry and cytotoxicity assay. Results: Here, we for the first time demonstrate that TiPSCs can effectively differentiate into functionally active NK cells without M210-B4 and AFT-024 xenogeneic stroma cells. Immunofluorescence and flow cytometry analysis showed that EB-derived cells can differentiate into a homogeneous population of NK cell expressing high levels of CD56, CD45 and CD16 specific markers. Moreover, these cells significantly express killing activation receptors such as NKp44 and NKp46. In the comparative analysis, we observed that NK cells derived using feeder-free culture system have more high killing activity against K-562 tumor cells, than NK cells derived by feeder-dependent method. Thus, we think that our obtained data will be useful for the development of large-scale production of NK cells for translation into cancer immunotherapy.

Keywords: induced pluripotent stem cells, NK cells, T cells, cell diffentiation, feeder cell-free culture system

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Authors: Sadia Arshad, Yifa Tang, Dumitru Baleanu

Abstract:

A fractional order mathematical model is proposed that incorporate CD8+ cells, natural killer cells, cytokines and tumor cells. The tumor cells growth in the absence of an immune response is modeled by logistic law as it was the simplest form for which predictions also agreed with the experimental data. Natural Killer Cells are our first line of defense. NK cells directly kill tumor cells through several mechanisms, including the release of cytoplasmic granules containing perforin and granzyme, expression of tumor necrosis factor (TNF) family members. The effect of the NK cells on the tumor cell population is expressed with the product term. Rational form is used to describe interaction between CD8+ cells and tumor cells. A number of cytokines are produced by NKs, including tumor necrosis factor TNF, IFN, and interleukin (IL-10). Source term for cytokines is modeled by Michaelis-Menten form to indicate the saturated effects of the immune response. Stability of the equilibrium points is discussed for biologically significant values of bifurcation parameters. We studied the treatment of fractional order system by investigating analytical conditions of tumor eradication. Numerical simulations are presented to illustrate the analytical results.

Keywords: cancer model, fractional calculus, numerical simulations, stability analysis

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Authors: Abdulbaset Mazarzaei

Abstract:

Natural killer (NK) cells are innate immune effectors that play a pivotal role in combating tumors and infected cells. In recent years, the therapeutic potential of NK cells has gained significant attention due to their remarkable cytotoxic ability. This study focuses on investigating the cytotoxic effect of expanded NK cells enriched with interleukin 12 (IL12) and interleukin 15 (IL15), derived from the leukoreduction filter, on the K562 cell line. Firstly, NK cells were isolated from whole blood samples obtained from healthy volunteers. These cells were subsequently expanded ex vivo using a combination of feeder cells, IL12, and IL15. The expanded NK cells were then harvested and assessed for their cytotoxicity against K562, a well-established human chronic myelogenous leukemia cell line. The cytotoxicity was evaluated using flow cytometry assay. Results demonstrate that the expanded NK cells significantly exhibited enhanced cytotoxicity against K562 cells compared to non-expanded NK cells. Interestingly, the expanded NK cells derived specifically from IL12 and IL15-enriched leukoreduction filters showed a robust cytotoxic effect similar to the whole blood-derived NK cells. These findings suggest that IL12 and IL15 in the leukoreduction filter are crucial in promoting NK cell cytotoxicity. Furthermore, the expanded NK cells displayed relatively similar cytotoxicity profiles to whole blood-derived NK cells, indicating their comparable capability in targeting and eliminating tumor cells. This observation is of significant relevance as expanded NK cells from the leukoreduction filter could potentially serve as a readily accessible and efficient source for adoptive immunotherapy. In conclusion, this study highlights the significant cytotoxic effect of expanded NK cells enriched with IL12 and IL15 obtained from the leukoreduction filter on the K562 cell line. Moreover, it emphasizes that these expanded NK cells exhibit comparable cytotoxicity to whole blood-derived NK cells. These findings reinforce the potential clinical utility of using expanded NK cells from the leukoreduction filter as an effective strategy in adoptive immunotherapy for the treatment of cancer. Further studies are warranted to explore the broader implications of this approach in clinical settings.

Keywords: natural killer (NK) cells, Cytotoxicity, Leukoreduction filter, IL-12 and IL-15 Cytokines

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Authors: Abobkar Saad, Qasmia Abdalla, Fatma Emhemed

Abstract:

Eighty-four of Calligonum comosum were cultured on Murashige and Skoog medium on every combination supplemented with different concentrations of IAA, BA, Zeatin, and GA3. When 84 seeds were inoculated on MS free hormones, different types of cells contain dense cytoplasm were observed ater 23 days and long thick wall cells arranged in layers. In case of using MS +BA(0.5mg/L), different types and shapes of parenchyma cells contain dense cytoplasm were detected after four weeks. In the case of using MS + BA(1mg/L) + GA3 (3mg/L), thick wall parenchyma cells contain dense cytoplasm after 19 days, but many layers of parenchyma cells contain dense cytoplasm after 28 days. When MS +kin(0.5mg/L) a thick cells wall as Sclereids were observed after 29 days. No any response were observed on Zeatin (0.5, 1 mg/L).

Keywords: anatomy, Calligonum comosum, in vitro, aeeds

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Authors: W. S. Li, S. T. Yang, H. C. Cheng

Abstract:

The power conversion efficiency (PCE) of the perovskite solar cells has been achieved by inserting vertically-aligned ZnO nanowires (NWs) between the cathode and the active layer and shows better solar cells performance. Perovskite solar cells have drawn significant attention due to the superb efficiency and low-cost fabrication process. In this experiment, ZnO nanowires are used as the electron transport layer (ETL) due to its low temperature process. The main idea of this thesis is utilizing the 3D structures of the hydrothermally-grown ZnO nanowires to increase the junction area to improve the photovoltaic performance of the perovskite solar cells. The infiltration and the surface coverage of the perovskite precursor solution changed as tuning the length of the ZnO nanowires. It is revealed that the devices with ZnO nanowires of 150 nm demonstrated the best PCE of 8.46 % under the AM 1.5G illumination (100 mW/cm2).

Keywords: hydrothermally-grown ZnO nanowires, perovskite solar cells, low temperature process, pinholes

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Authors: Chakrit Thapphan, Suteeraporn Chaowattanapanit, Sorutsiri Chareonsudjai, Wisitsak Phoksawat, Supranee Phantanawiboon, Kiatichai Faksri, Steve W. Edwards, Kanin Salao

Abstract:

Background: Psoriasis is a chronic inflammatory disease of the skin that is mediated by crosstalk between keratinocytes and immune cells, especially CD4+ T helper (Th) cells. To date, psoriasis is established as a T helper 17 (Th17) cell-mediated inflammatory process driven by the over-expression of Th17. However, the role of other CD4+T helper cells is rather controversial. Objective: Our study, thereby, aimed to characterize and analyze T cell subsets in the circulating blood of psoriasis patients and compare them to healthy controls. Methods: Peripheral blood mononuclear cells were isolated from the participants and stained with fluorescent dye-conjugated monoclonal antibodies specific for intracellular cytokines, including interferon-gamma (IFN- γ), interleukin (IL-4), IL-17 and forkhead box P3 (FOXP3), that can be used to define T helper 1 (Th1) cells, T helper 2 (Th2), T helper 17 (Th17) and regulatory T cells (Treg) respectively. Results: We found that the numbers of Th2 (59.6% ± 17.0) and Th17 (4.0% ± 2.0) cells in the circulating blood of psoriasis patients were significantly higher than those of the healthy controls (p= 0.0007 and 0.0013 respectively). In contrast, the numbers of Th1 and Treg cells were not significantly different between psoriasis patients and healthy controls (p= 0.0593 and 0.8518, respectively). Additionally, when adjusting these numbers of Th cells to Treg, we observed a similar trend that the ratio of Th2/Treg and Th17/Treg also elevated (p = 0.0007 and 0.0047, respectively). Conclusion: Taken together, our results suggest an imbalanced T exhibit toward the Th2 and Th17 skewed-immune responses in psoriasis patients.

Keywords: psoriasis, Th cell subsets, Th2 cells, Th17 cells, Treg cells

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Authors: Kirsten Wilson, Patrick M. Brauer, Sandra Babic, Diana Golubeva, Jessica Van Eyk, Tinya Wang, Avanti Karkhanis, Tim A. Le Fevre, Andy I. Kokaji, Allen C. Eaves, Sharon A. Louis, , Nooshin Tabatabaei-Zavareh

Abstract:

Long-lived plasma cells are rare, non-proliferative B cells generated from antibody-secreting cells (ASCs) following an immune response to protect the host against pathogen re-exposure. Despite their therapeutic potential, the lack of in vitro protocols in the field makes it challenging to use B cells as a cellular therapeutic tool. As a result, there is a need to establish robust and reproducible methods for the generation of B cells. To address this, we have developed a culture system for generating B cells from hematopoietic stem and/or progenitor cells (HSPCs) derived from human umbilical cord blood (CB) or pluripotent stem cells (PSCs). HSPCs isolated from CB were cultured using the StemSpan™ B Cell Generation Kit and produced CD19+ B cells at a frequency of 23.2 ± 1.5% and 59.6 ± 2.3%, with a yield of 91 ± 11 and 196 ± 37 CD19+ cells per input CD34+ cell on culture days 28 and 35, respectively (n = 50 - 59). CD19+IgM+ cells were detected at a frequency of 31.2 ± 2.6% and were produced at a yield of 113 ± 26 cells per input CD34+ cell on culture day 35 (n = 50 - 59). The B cell receptor loci of CB-derived B cells were sequenced to confirm V(D)J gene rearrangement. ELISpot analysis revealed that ASCs were generated at a frequency of 570 ± 57 per 10,000 day 35 cells, with an average IgM+ ASC yield of 16 ± 2 cells per input CD34+ cell (n = 33 - 42). PSC-derived HSPCs were generated using the STEMdiff™ Hematopoietic - EB reagents and differentiated to CD10+CD19+ B cells with a frequency of 4 ± 0.8% after 28 days of culture (n = 37, 1 embryonic and 3 induced pluripotent stem cell lines tested). Subsequent culture of PSC-derived HSPCs increased CD19+ frequency and generated ASCs from 1 - 2 iPSC lines. This method is the first report of a serum- and feeder-free system for the generation of B cells from CB and PSCs, enabling further B lineage-specific research for potential future clinical applications.

Keywords: stem cells, B cells, immunology, hematopoiesis, PSC, differentiation

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Authors: Hadis Pouyafar, D. Matin Alaghmandan

Abstract:

Gallium arsenide (GaAs) solar cells are widely used in applications like spacecraft and satellites because they have a high absorption coefficient and efficiency and can withstand high-energy particles such as electrons and protons. With the energy crisis, there's a growing need for efficiency and cost-effective solar cells. GaAs cells, with their 46% efficiency compared to silicon cells 23% can be utilized in buildings to achieve nearly zero emissions. This way, we can use irradiation and convert more solar energy into electricity. III V semiconductors used in these cells offer performance compared to other technologies available. However, despite these advantages, Si cells dominate the market due to their prices. In our study, we took an approach by using software from the start to gather all information. By doing so, we aimed to design the optimal building that harnesses the full potential of solar energy. Our modeling results reveal a future; for GaAs cells, we utilized the Grasshopper plugin for modeling and optimization purposes. To assess radiation, weather data, solar energy levels and other factors, we relied on the Ladybug and Honeybee plugins. We have shown that silicon solar cells may not always be the choice for meeting electricity demands, particularly when higher power output is required. Therefore, when it comes to power consumption and the available surface area for photovoltaic (PV) installation, it may be necessary to consider efficient solar cell options, like GaAs solar cells. By considering the building requirements and utilizing GaAs technology, we were able to optimize the PV surface area.

Keywords: gallium arsenide (GaAs), optimization, sustainable building, GaAs solar cells

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Authors: Sumayah Mohammed Asiri A., Aviva Levina B., Elizabeth New C., Peter Lay D.

Abstract:

Pt compounds have been among the most successful anticancer drugs in the last 40 years, but the development of resistance to them is an increasing problem. Cellular homeostasis of an essential metal, Cu, is known to be involved in Pt resistance, but mechanisms of this process are poorly understood. We used a novel ratiometric Cu(I)-sensitive fluorescent probeInCCu1 dye to detect Cu(I) in the mitochondria. Total Cu and labile Cu pool measured using AAS and InCCu1 dye in A2780 cells and their corresponding resistant cells A2780-cis.R cells treated with Cu and cisplatin. The main difference between both cell lines in the presence and absence of Cu(II) is that resistant cells have lower total Cu content but higher labile Cu levels than cisplatin-sensitive cells. This means that resistant cells can metabolize and export excess Cu more efficiently. Furthermore, InCCu1 has emerged not only as an indicator of labile cellular Cu levels in the mitochondria but as a potentially versatile multi-organelle probe.

Keywords: AAS and ICPMS, A2780 and its resistant cells, ratiometric fluorescent sensors, inCCu1, and total and labile Cu

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