Search results for: primary porcine respiratory epithelial cells (PoRECs)
7604 E-teaching Barriers: A Survey from Shanghai Primary School Teachers
Authors: Liu Dan
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It was considered either unnecessary or impossible for primary school students to implement online teaching until last year. A large number of E-learning or E-teaching researches have been focused on adult-learners, andragogy and technology, however, primary school education, it is facing many problems that need to be solved. Therefore, this research is aimed at exploring barriers and influential factors on online teaching for K-12 students from teachers’ perspectives and discussing the E-pedagogy that is suitable for primary school students and teachers. Eight hundred and ninety-six teachers from 10 primary schools in Shanghai were invited to participate in a questionnaire survey. Data were analysed by hierarchical regression, and the results stress the significant three barriers by teachers with online teaching: the existing system is deficient in emotional interaction, teachers’ attitude towards the technology is negative and the present teacher training is lack of systematic E-pedagogy guidance. The barriers discovered by this study will help the software designers (E-lab) develop tools that allow for flexible and evolving pedagogical approaches whilst providing an easy entry point for cautious newcomers, so that help the teachers free to engage in E-teaching at pedagogical and disciplinary levels, to enhance their repertoire of teaching practices.Keywords: online teaching barriers (OTB), e-teaching, primary school, teachers, technology
Procedia PDF Downloads 2017603 The Impact of Total Dust (LGS) and Mineral Dust (PM 10) in Cardio Vascular and Respiratory System, in Albania: A Longitudinal Study
Authors: Canga Mimoza, Irene Malagnino, Giulia Malagnino, Vito Malagnino
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Aim: This study aims at evaluating the impact of total dust (LGS) and mineral dust (PM10), in the cardio vascular and respiratory systems. Also proving that these air polluters are the cause of several diseases, such as bronchopneumonia, pneumonia, bronchitis, angina pectoris and cardiac insufficiency. Material and Method: The study is concentrated in the cities of Fier and Vlora. This is a clinic-epidemiological study conducted during the time period 2014-2019. Some of the data of LGS and PM10 were obtained from the database of the Institute of Public Health. The formula to measure the mean value of LGS and PM10 is ∆X=X (mean)-Xᵢ. Results: Based on the calculations made, we noticed that: The mean value of LGS in the city of Fieri was 227,33, while the mean value of LGS in the city of Vlora was 177,4. Whereas, the mean value of PM10 in the city of Fieri was 105.5 and the mean value of PM10 in the city of Vlore was 77.5. According to, our statistics the values of LGS were 1.2 times higher in Fier than in Vlora and the PM10 values were 1.36 times higher in Fier than in Vlora. Based on the data, in the city of Fier, the incidence of the bronchopneumonia was 56.53 sick patients/1000 inhabitants, but in Vlora, it was 22 sick patients/1000 inhabitants, so the number of the sick patients was 2.5 times higher in the city of Fieri compared with Vlora city, (P=0.001). The number of the patients with bronchitis, in the city of Fier, was 18 patients/1000 inhabitants, whereas, in Vlora, it was 9 patients/1000 inhabitants, (P=0.005). Based on the data, 8 patients/1000 inhabitants in the city of Fier, suffered from the pneumonia disease, while in Vlora city, were 4 patients/1000 inhabitants, (P=0.005). Another disease taken in consideration was angina pectoris. This study can claim that in the city of Fier, 9.5 patients/1000 inhabitants suffered from this disease, while in Vlora city, were only 4 patients /1000 inhabitants, (P=0.001). Findings of the present study proved that 3.7 patients/1000 inhabitants in the city of Fieri, had cardiac insufficiency, whereas in the city of Vlora, were 1.8 patients/1000 inhabitants, (P=0.05). Conclusions: LGS and PM10 have an influential impact on the cardio vascular and respiratory system; that’s why their levels should be kept under control. The pollution levels are 1.2 and 1.4 times higher in Fier than in Vlora; also the incidences of the diseases are 2 times higher in Fier than in Vlora. Recommendations: In order to prevent the cardio vascular and respiratory diseases, we should avoid places where pollution is higher than the norm. This can be achieved by frequenting places where the air pollution is lower, such as parks, gardens, top floors, etc.Keywords: impact of total dust, LGS, mineral dust, PM 10, cardio vascular pathologies, respiratory disease
Procedia PDF Downloads 1287602 Characterization of Retinal Pigmented Cell Epithelium Cell Sheet Cultivated on Synthetic Scaffold
Authors: Tan Yong Sheng Edgar, Yeong Wai Yee
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Age-related macular degeneration (AMD) is one of the leading cause of blindness. It can cause severe visual loss due to damaged retinal pigment epithelium (RPE). RPE is an important component of the retinal tissue. It functions as a transducing boundary for visual perception making it an essential factor for sight. The RPE also functions as a metabolically complex and functional cell layer that is responsible for the local homeostasis and maintenance of the extra photoreceptor environment. Thus one of the suggested method of treating such diseases would be regenerating these RPE cells. As such, we intend to grow these cells using a synthetic scaffold to provide a stable environment that reduces the batch effects found in natural scaffolds. Stiffness of the scaffold will also be investigated to determine the optimal Young’s modulus for cultivating these cells. The cells will be generated into a monolayer cell sheet and their functions such as formation of tight junctions and gene expression patterns will be assessed to evaluate the cell sheet quality compared to a native RPE tissue.Keywords: RPE, scaffold, characterization, biomaterials, colloids and nanomedicine
Procedia PDF Downloads 4357601 Genome Sequencing of Infectious Bronchitis Virus QX-Like Strain Isolated in Malaysia
Authors: M. Suwaibah, S. W. Tan, I. Aiini, K. Yusoff, A. R. Omar
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Respiratory diseases are the most important infectious diseases affecting poultry worldwide. One of the avian respiratory virus of global importance causing significant economic losses is Infectious Bronchitis Virus (IBV). The virus causes a wide spectrum disease known as Infectious Bronchitis (IB), affecting not only the respiratory system but also the kidney and the reproductive system, depending on its strain. IB and Newcastle disease are two of the most prevalent diseases affecting poultry in Malaysia. However, a study on the molecular characterization of Malaysian IBV is lacking. In this study, an IBV strain IBS130 which was isolated in 2015 was fully sequenced using next-gene sequencing approach. Sequence analysis of IBS130 based on the complete genome, polyprotein 1ab and S1 genes were compared with other IBV sequences available in Genbank, National Center for Biotechnology Information (NCBI). IBV strain IBS130 is characterised as QX-like strain based on whole genome and S1 gene sequence analysis. Comparisons of the virus with other IBV strains showed that the nucleotide identity ranged from 67% to 99.2%, depending on the region analysed. The similarity in whole genome nucleotide ranging from 84.9% to 90.7% with the least similar was from Singapore strains (84.9%) and highly similar with China QX-like strains. Meanwhile, the similarity in polyprotein 1ab ranging from 85.3% to 89.9% with the least similar to Singapore strains (85.3%) and highly similar with Mass strains from USA.Keywords: infectious bronchitis virus, phylogenetic analysis, chicken, Malaysia
Procedia PDF Downloads 1867600 The Effect of Manggong Bamboo Leaves Extract (Gigantochloa manggong) on Rat (Rattus novergicus) Blood Profile
Authors: Sri Rahayu, Supriyatin, Yuli Rahma Dini
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One of the consequences of excess physical activity is the oxidative stress which resulted in damage to blood cells. Oxidative stress condition can be reduced by an exogenous antioxidant. The natural exogenous antioxidant can be extracted from Manggong bamboo (Gigantochloa manggong). This research was aim to evaluate the effect of physical exercise and Manggong bamboo (Gigantochloa manggong) leaf extract on blood profile of rats. This research was conducted in July 2013 to May 2014 using experimental method with completely randomized design (CRD) with two factors, physical exercise and Manggong bamboo leaf extract. The rats blood profile to be measured were the level of erythrocyte cells, leucocyte cells and hemoglobin. Data were analyzed with parametric statistical 2-way ANOVA test (α = 0.05). Manggong bamboo leaf extract was non toxic and contained flavonoid, triterpenoid, saponin and alkaloid. There was an effect of physical exercise and manggong bamboo leaf extract on blood profile of rats. Data obtained on physical activity, giving erythrocyte cells (2.5 million/µl) and hemoglobin (12,42g/dL) declined compared to the number of leucocyte cells increases (6,500cells/L). Extract treatment was increased the erythrocytes (5,13 million/µl) and hemoglobin level (14,72 g/dL.) while the leukocytes level were decreased (1.591,67 cells/L). The extract and physical activity treatment showed an increase in erythrocytes (2,96 million/µl) and hemoglobin (14,3 g/dL) but decrease the number of leukocytes (1.291,67 cells/L). The conclusion was that physical activity and Manggong bamboo leafs extract gaves effect on the blood profile of white rat.Keywords: antioxidant, blood profile of rats, Manggong bamboo leaf extract, leukocytes
Procedia PDF Downloads 2907599 The Abnormality of Blood Cells Parasitized by Plasmodium vivax
Authors: Manas Kotepui, Kwuntida Uthaisar, Phiman Thirarattanasunthon, Bhukdee PhunPhuech, Nuoil Phiwklam
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Introduction: Malaria due to Plasmodium vivax has placed huge burdens on the health, longevity, and general prosperity of large sections of the human population. This study aimed at prospectively collecting information on the clinical profile of Plasmodium vivax from subjects acutely infected with P. vivax residing in some of the highest malaria transmission regions in Thailand. Methods: A retrospective study of malaria cases, hospitalized between 2013 and 2015 was performed. Clinical characteristics, diagnosis, and parasitological results on admission, age, and gender were mined from medical records at Phop Phra Hospital located in endemic areas of Tak Province, Thailand. Venous blood samples were collected at the time of admission to the hospital to determine the present of parasite and also parasite count by thick and thin film examination, and also Complete blood count (CBC) parameters. Results: Results showed that patients infected with Plasmodium vivax (276 cases) had a high monocyte count (mean=390 cells/µL) during initial stage of infection and continuously lower during later stage (any stage with gametocyte, mean=230 cells/µL) of infection (P value=0.021) whereas, patients infected with Plasmodium vivax had a low basophil count (mean=20 cells/µL) during initial stage of infection and continuously higher during later stage of infection (mean at stage with gametocyte=70 cells/µL) (P value=0.033). In addition, patients with more than one stage infection tend to have lower lymphocyte count (mean=1180 cells/µL) than patients with only one stage infection (mean=1350 cells/µL)(P value=0.011) whereas, patients with more than one stage infection tend to have lower basophil count (mean=60 cells/µL) than patients with only one stage infection (mean=80 cells/µL) (P value=0.01). Conclusion: This study indicated that patients infected with Plasmodium vivax had high monocyte count and low basophil count during initial stage of infection which was continuously lower during later stage of infection. Patients with more than one stage infection tend to have lower lymphocyte count than patients with only one stage infection whereas, patients with more than one stage infection tend to have lower basophil count than patients with only one stage infection. This information contributes to better understanding of pathological characteristic of Plasmodium vivax infection.Keywords: plasmodium vivax, Thailand, asexual erythrocytic stages, hematological parameters
Procedia PDF Downloads 2127598 The Relationship between Human Neutrophil Elastase Levels and Acute Respiratory Distress Syndrome in Patients with Thoracic Trauma
Authors: Wahyu Purnama Putra, Artono Isharanto
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Thoracic trauma is trauma that hits the thoracic wall or intrathoracic organs, either due to blunt trauma or sharp trauma. Thoracic trauma often causes impaired ventilation-perfusion due to damage to the lung parenchyma. This results in impaired tissue oxygenation, which is one of the causes of acute respiratory distress syndrome (ARDS). These changes are caused by the release of pro-inflammatory mediators, plasmatic proteins, and proteases into the alveolar space associated with ongoing edema, as well as oxidative products that ultimately result in severe inhibition of the surfactant system. This study aims to predict the incidence of acute respiratory distress syndrome (ARDS) through human neutrophil elastase levels. This study examines the relationship between plasma elastase levels as a predictor of the incidence of ARDS in thoracic trauma patients in Malang. This study is an observational cohort study. Data analysis uses the Pearson correlation test and ROC curve (receiver operating characteristic curve). It can be concluded that there is a significant (p= 0.000, r= -0.988) relationship between elastase levels and BGA-3. If the value of elastase levels is limited to 23.79 ± 3.95, the patient will experience mild ARDS. While if the value of elastase levels is limited to 57.68 ± 18.55, in the future, the patient will experience moderate ARDS. Meanwhile, if the elastase level is between 107.85 ± 5.04, the patient will likely experience severe ARDS. Neutrophil elastase levels correlate with the degree of severity of ARDS incidence.Keywords: ARDS, human neutrophil elastase, severity, thoracic trauma
Procedia PDF Downloads 1487597 Mannosidase Alpha Class 1B Member 1 Targets F Severe Acute Respiratory Syndrome Coronavirus 2 Spike Protein and Ebola Virus Glycoprotein to Endoplasmic Reticulum-To-Lysosome-Associated Degradation by Micro-Endoplasmic Reticulum-Phagy
Authors: Yong-Hui Zheng
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Viruses hijack host machineries to propagate and spread, which disrupts cellular homeostasis and activates various counteractive mechanisms. Infection of enveloped viruses is dependent on their fusion proteins, which bind to viral receptors to allow virus entry into cells. Fusion proteins are glycoproteins and expressed in the endoplasmic reticulum (ER) by hijacking the secretory pathway. Previously, we reported that Zaire ebolavirus (EBOV)-glycoprotein (GP) expression induces ER stress, and EBOV-GP is targeted by the calnexin cycle to macro-ER-phagy for degradation. We now report that expression of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2/SARS2)-spike (S) protein also causes ER stress, and its expression is strongly downregulated by mannosidase alpha class 1B member 1 (MAN1B1), a class I α-mannosidase from the ER. MAN1B1 co-localizes with SARS2-S in the ER, and its downregulation of SARS2-S is blocked by inhibitors targeting lysosomes and autophagy, but not proteasomes, indicating SARS2-S degradation by autolysosomes. Notably, the SARS2-S degradation does not require the core autophagy machinery including ATG3, ATG5, ATG7, and phosphatidylinositol 3-kinase catalytic subunit type 3 (PI3KC3)/vacuolar protein sorting 34 (VPS34), and instead, it requires Beclin 1 (BECN1), a core component in the PI3KC3 complex. In addition, MAN1B1 does not trigger SARS2-S polyubiquitination, and consistently, the SARS2-S degradation does not require the autophagy receptor sequestosome 1 (SQSTM1)/p62. MAN1B1 also downregulates EBOV-GP similarly, but this degradation does not require BECN1. Collectively, we conclude that MAN1B1 downregulates viral fusions by micro-ER-phagy, and importantly, we have identified BECN1-dependent and BECN1-independent mechanisms for micro-ER-phagy.Keywords: Micro-ER-phagy, reticulophagy, fusion proteins, ER stress
Procedia PDF Downloads 697596 Supergranulation and Its Turbulent Convection
Authors: U. Paniveni
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A few parameters of supergranular cells are studied using intensity patterns from the Kodaikanal Solar Observatory and Dopplergrams from SOHO. The turbulent aspect of the solar supergranulation is established by examining the interrelationships amongst the parameters characterizing a supergranular cell, namely size, lifetime, area, perimeter, fractal dimension, and horizontal flow velocity. The complexity of supergranular cells depicted by their fractal dimension is indicative of their non-laminar characteristics. The findings corroborate Kolmogorov’s theory of turbulence. Some parameters of supergranular cells also show a latitudinal dependence. Supergranulation is a synonym of convective phenomenon and hence can shed light on the physical conditions in the convection zone of the Sun. It plays a major role in the transport and dispersal of magnetic fields that may have a relation to the phases of the solar cycle.Keywords: sun, granulation, convection, turbulence
Procedia PDF Downloads 407595 Effector and Memory Immune Responses Induced by Total Extracts of Naegleria fowleri Co-Administered with Cholera Toxin
Authors: Q. B. Maria de la Luz Ortega Juárez, Saúl Rojas Hernández, Itzel Berenice Rodríguez Mera, María Maricela Carrasco Yépez, Mara Gutierrez Sánchez
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Naegleria fowleri is a free-living amoeba found mainly in temperate freshwater and is the etiologic agent of primary amebic meningoencephalitis (PAM), a fatal acute disease with a mortality rate greater than 95%. At present, there are no treatments available for MAP, and the development of effective vaccines that generate long-term immunological memory allowing protection against MAP would be of great importance. The objective of this work was to analyze the effector and memory immune response in BALB/c mice immunized with total extract of N. fowleri co-administered with cholera toxin. In this study, BALB/c mice were immunized four times intranasally with ET of N. fowleri adjuvanted with CT with or without booster at three months and were challenged or not with the lethal dose of N. fowleri, determining survival, the humoral, effector and memory response, by ELISA and flow cytometry techniques. The results obtained showed that the survival of mice immunized with booster had 60% protection compared to the group without booster, which obtained 20% protection. Evaluating the humoral response, it was found that both IgG and IgA levels were higher in sera than in nasal washes in both treatments. In the cellular response, the increase in the percentage of positive cells was found for effector T and B lymphocytes in the nasal passages (NP) in the group with boost and nasopharynx-associated lymphoid tissue (NALT) in the group without boost and lymphocytes only. B in both treatments, as well as in memory cells treatment with boost T lymphocytes in PN and NALT and without boost in cervical lymph nodes (CG) with respect to B lymphocytes, in PN, GC and NALT in treatment with boost and NALT in treatment without booster. Therefore, the involvement of the effector immune response and memory play a fundamental role for protection against N. fowleri and for the development of vaccine candidates.Keywords: immune response, immunological memory, naegleria fowleri, primary amebic meningoencephalitis
Procedia PDF Downloads 787594 Calpain-Mediated, Cisplain-Induced Apoptosis in Breast Cancer Cells
Authors: Shadia Al-Bahlani, Khadija Al-Bulushi, Zuweina Al-Hadidi, Buthaina Al-Dhahl, Nadia Al-Abri
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Breast cancer is the most common cancer in women worldwide. Triple-Negative Breast Cancer (TNBC) is an aggressive type of breast cancer, which is defined by the absence of Estrogen (ER), Progesterone (PR) and human epidermal growth factor (Her-2) receptors. The calpain system plays an important role in many cellular processes including apoptosis, necrosis, cell signaling and proliferation. However, the role of calpain in cisplatin (CDDP)-induced apoptosis in TNBC cells is not fully understood. Here, TNBC (MDA-MB231) cells were treated with different concentration of CDDP (0, 20 & 40 µM) and calpain activation and apoptosis were measured by western blot and Hoechst Stain respectively. In addition, calpain modulation by either activation and/or inhibition and its effect on CDDP-induced apoptosis were assessed by the same above approaches. Our findings showed that CDDP induced endoplasmic reticulum stress and thus Calcium release and subsequently activate calpain α-fodrin cleavage indicated by the increase in GRP78 and Calmodulin protein expression and respectively in MDA-MB231 cells. It also induced apoptosis as measured by Hoechst stain and caspase-12 cleavage. Calpain activation by both Cyclopiazonic acid and Thapsigargin showed similar effect and enhanced the sensitivity of these cells to CDDP treatment. On the other hand, calpain inhibition by either specific siRNA and/or exogenous inhibitor (Calpeptin) had an adverse effect where it attenuated calpain activation and thus CDDP- induced apoptosis in these cells. Altogether, these findings suggested that calpain activation play an essential role in sensitizing the TNBC cells to CDDP-induced apoptosis. This might lead to the discovery of novel treatment to over this aggressive type of breast cancer.Keywords: calpain, cisplatin, apoptosis, breast cancer
Procedia PDF Downloads 3457593 Case Report: Rare Case of Endometrial Stromal Sarcoma with Omental Metastasis in a 19-Year Old Girl
Authors: Mukurdipi Ray, Seema Singh
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Extrauterine endometrial stromal sarcoma (ESS) is a rare entity and typified by delayed recurrence of primary ESS. Here, we present an unusual case of uterine ESS in a woman with a history of hysterectomy. A 19-year-old girl, underwent a hysterectomy and bilateral salpingo-oophorectomy for uterine ESS 12 months ago and now after remaining disease free for nine months ago she presented with ascites along with pelvic and peritoneal mass. Intraoperatively, the large omental mass was found, and optimal cytoreduction with total omentomy (supracolic and infracolic ) total peritonectomy and hyperthermic intraperitoneal chemotherapy (HIPEC) was offered to the patient. Final histopathology report showed the involvement of only omentum by ESS cells. Immunohistochemistry (IHC) and receptor study were done and it was positive for CD-10 and desmin and negative for CK- 7. This case highlights the rarity of extrauterine ESS in the omentum with a known history of primary uterine ESS which was treated successfully with the above-mentioned procedure. Though active and long-term surveillance is recommended to monitor for late recurrences.Keywords: endrometrial stromal sarcoma, complete cytoreduction, hyperthermic intra peritoneal chemotherapy, total omentectomy
Procedia PDF Downloads 2077592 Influence of Physico-Chemical Changes in the Environment on the Behavior of Tadpoles Rana Saharica: Case of Fungicide (Artea 330ec)
Authors: S. Zouainia, R. Djebar, H. Berrebah, A. Sayeb
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This work focused on the study of physiological and biochemical changes observed in tadpoles exposed to fungicide Rana saharica Artea 330ec recently introduced in Algeria. For this, we tested the effect of xenobiotics on growth and development of tadpoles; among the studied parameters: total protein, glutathione and respiratory activity. The study of physiological parameters showed that the tadpoles change perfectly in the absence of toxic and in favorable conditions (pH, temperature). Our results showed an increased rate of protein and GSH in the presence of the fungicide Artea 330ec. The latter causes uninhibited very highly significant respiratory activity of tadpoles treated. The presence of xenobiotics in the breeding tadpoles water causes disturbances in behavior and food metabolism.Keywords: amphibians, fungicides, pesticides, pollution
Procedia PDF Downloads 4087591 Microdosimetry in Biological Cells: A Monte Carlo Method
Authors: Hamidreza Jabal Ameli, Anahita Movahedi
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Purpose: In radionuclide therapy, radioactive atoms are coupled to monoclonal antibodies (mAbs) for treating cancer tumor while limiting radiation to healthy tissues. We know that tumoral and normal tissues are not equally sensitive to radiation. In fact, biological effects such as cellular repair processes or the presence of less radiosensitive cells such as hypoxic cells should be taken account. For this reason, in this paper, we want to calculate biological effect dose (BED) inside tumoral area and healthy cells around tumors. Methods: In this study, deposited doses of a radionuclide, gold-198, inside cells lattice and surrounding healthy tissues were calculated with Monte Carlo method. The elemental compositions and density of malignant and healthy tissues were obtained from ICRU Report 44. For reaching to real condition of oxygen effects, the necrosis and hypoxia area inside tumors has been assessed. Results: With regard to linear-quadratic expression which was defined in Monte Carlo, results showed that a large amount of BED is deposited in the well-oxygenated part of the hypoxia area compared to necrosis area. Moreover, there is a significant difference between the curves of absorbed dose with BED and without BED.Keywords: biological dose, monte carlo, hypoxia, radionuclide therapy
Procedia PDF Downloads 4877590 Contraction and Membrane Potential of C2C12 with GTXs
Authors: Bayan Almofty, Yuto Yamaki, Tadamasa Terai, Sadahito Uto
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Culture techniques of skeletal muscle cells are advanced in the field of regenerative medicine and applied research of cultured muscle. As applied research of cultured muscle, myopathy (muscles disease) treatment is expected and development bio of actuator is also expected in biomedical engineering. Grayanotoxins (GTXs) is known as neurotoxins that enhance the permeability of cell membrane for Na ions. Grayanotoxins are extracted from a famous Pieris japonica and Ericaceae as well as a phytotoxin. In this study, we investigated the effect of GTXs on muscle cells (C2C12) contraction and membrane potential. Contraction of myotubes is induced by applied external electrical stimulation. Contraction and membrane potential change of skeletal muscle cells are induced by injection of current. We, therefore, concluded that effect of Grayanotoxins on contraction and membrane potential of C2C12 relate to acute toxicity of GTXs.Keywords: skeletal muscle cells C2C12, grayanotoxins, contraction, membrane potential, acute toxicity, pytotoxin, motubes
Procedia PDF Downloads 5057589 Comparative Analysis of Teachers’ Performance in Private and Public Primary Schools in Oyo State
Authors: Babajide Solomon Faloore
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This study on the comparative analysis of the performance of teachers in private and public schools was carried out in Ibadan North West Local Government Area of Oyo State. This study examined the justification for the claim that there is a difference in the performance of teachers in private and public primary schools and at the same time identified factors responsible for the difference in the performance of these teachers. A descriptive survey research design was used for the study. Data generated were analyzed using t-test and regression analysis. The findings of the study revealed that there is significance difference in the performance of teachers in private and private primary schools in Ibadan North West Local Government Area of Oyo State( t=64.09; df=459; p,.05). The findings also revealed that the method of teaching in private primary schools is significantly different from the method of teaching in public primary schools in Ibadan North West Local Government Area of Oyo State (t=73.08; df=459; p,.05). Findings revealed that school leadership and management have a significant contribution on the performance of private and public primary school teachers in Ibadan North West Local Area of Oyo State. Based on the finding, the following recommendations were made: Primary school teachers need to be motivated and rewarded for excellent performance. Primary schools should be properly equipped with teaching–aid facilities, laboratories, and libraries. The government should use the findings of this study to improve on teaching materials provided to the primary school teachers in Nigeria. Public primary schools should be designed by education planners, administrators, and government. Headmasters, proprietors, and teachers of primary schools should look inward and give a performance appraisal and evaluation of themselves from time to time based on the subject they taught. Finally, school administrators should be conscious of the way they manage the teachers in schools not only in informal situations but also in formal settings.Keywords: private education, public education, school leadership, school management, teachers performance
Procedia PDF Downloads 3907588 Fabrication and Properties of Al2O3/Si Quantum Well-Structured Silicon Solar Cells
Authors: Kwang-Ho Kim, Kwan-Hong Min, Pyungwoo Jang, Chisup Jung, Kyu Seomoon
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By restricting the dimensions of silicon to less than Bohr radius of bulk crystalline silicon (∼5 nm), quantum confinement causes its effective bandgap to increase. Therefore, silicon quantum wells (QWs) using these quantum phenomena could be a good candidate to achieve high performance silicon solar cells. The Al2O3/Si QW structures were fabricated by using the successive deposition technique, as a quantum confinement device to increase the effective energy bandgap and passivation effect in Si surface for the 3rd generation solar cell applications. In Si/Al2O3 QWs, the thicknesses of Si layers and Al2O3 layers were varied between 1 to 5 nm, respectively. The roughness of deposited Si on Al2O3 was less than 4 Å in the thickness of 2 nm. By using the Al2O3/Si QW structures on Si surfaces, the lifetime measured by u-PCD technique increased as a result of passivated surface effects. The discussion about the other properties such as electrical and optical properties of the QWs structures as well as the fabricated solar cells will be presented in this paper.Keywords: Al2O3/Si quantum well, quantum confinement, solar cells, third generation, successive deposition technique
Procedia PDF Downloads 3407587 Anatomically-Based Oropharyngeal Rehabilitation for the Patients with Obstructive Sleep Apnea Using a Multilevel Approach
Authors: Hsin-Yu Lin, Ching-Hsia Hung
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Obstructive sleep apnea (OSA) is characterized by a complete or partial obstruction of the upper airway during sleep. The vulnerable sites of upper airway collapses are consequences of sleep state-dependent reductions in tone in specific pharyngeal dilators. Clinical examinations reveal multilevel collapses of the upper airway among the patients with OSA. Therefore, an anatomically-based oropharyngeal rehabilitation should comprise a multilevel approach, including retropalatal, retroglossal, hypopharyngeal, temporomandibular, and facial levels, all of which involve different muscle groups and contribute to multifunctional interaction and coordination, such as swallowing, breathing, and phonation. The purpose of the study was to exam the effects of this rehabilitation program with a multilevel approach. In this study, fifteen subjects with newly diagnosed moderate or severe OSA (Apnea-Hypopnea-Index≥15) were randomized into an intervention group and control group. The intervention group (N=8) underwent a 12-week-intervention of a hospital-based rehabilitation program, while the control group (N=7) was kept on the waiting list. The 12-week-intervention comprised an anatomically based multilevel approach. The primary outcome was Polysomnography (PSG) data, and the secondary outcome was oropharyngeal and respiratory muscle function. In the intervention group, Apnea-Hypopnea-Index significantly improved (46.96±19.45 versus 32.78±10.78 events/h, p=0.017) compared with control group (35.77±17.49 versus 42.96±17.32 events/h, p=0.043). While the control group remained no change, the intervention group demonstrated other PSG outcomes significantly improvement, including arousal index (46.04±18.9 versus 32.98±8.35/h, p=0.035), mean SpO2 (92.88±2.1 versus 94.13±1.46%, p=0.039). Besides, the intervention group demonstrated significant improvement in oropharyngeal and respiratory muscle function compared to the control group. This anatomically-based oropharyngeal rehabilitation with a multilevel approach can be proven as a non-invasive therapy for patients with OSA.Keywords: obstructive sleep apnea, upper airway, oropharyngeal rehabilitation, multilevel approach
Procedia PDF Downloads 1027586 Spectral Response Measurements and Materials Analysis of Ageing Solar Photovoltaic Modules
Authors: T. H. Huang, C. Y. Gao, C. H. Lin, J. L. Kwo, Y. K. Tseng
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The design and reliability of solar photovoltaic modules are crucial to the development of solar energy, and efforts are still being made to extend the life of photovoltaic modules to improve their efficiency because natural aging is time-consuming and does not provide manufacturers and investors with timely information, accelerated aging is currently the best way to estimate the life of photovoltaic modules. In this study, the accelerated aging of different light sources was combined with spectral response measurements to understand the effect of light sources on aging tests. In this study, there are two types of experimental samples: packaged and unpackaged and then irradiated with full-spectrum and UVC light sources for accelerated aging, as well as a control group without aging. The full-spectrum aging was performed by irradiating the solar cell with a xenon lamp like the solar spectrum for two weeks, while the accelerated aging was performed by irradiating the solar cell with a UVC lamp for two weeks. The samples were first visually observed, and infrared thermal images were taken, and then the electrical (IV) and Spectral Responsivity (SR) data were obtained by measuring the spectral response of the samples, followed by Scanning Electron Microscopy (SEM), Raman spectroscopy (Raman), and X-ray Diffraction (XRD) analysis. The results of electrical (IV) and Spectral Responsivity (SR) and material analyses were used to compare the differences between packaged and unpackaged solar cells with full spectral aging, accelerated UVC aging, and unaged solar cells. The main objective of this study is to compare the difference in the aging of packaged and unpackaged solar cells by irradiating different light sources. We determined by infrared thermal imaging that both full-spectrum aging and UVC accelerated aging increase the defects of solar cells, and IV measurements demonstrated that the conversion efficiency of solar cells decreases after full-spectrum aging and UVC accelerated aging. SEM observed some scorch marks on both unpackaged UVC accelerated aging solar cells and unpackaged full-spectrum aging solar cells. Raman spectroscopy examines the Si intensity of solar cells, and XRD confirms the crystallinity of solar cells by the intensity of Si and Ag winding peaks.Keywords: solar cell, aging, spectral response measurement
Procedia PDF Downloads 1017585 Global Stability Analysis of a Coupled Model for Healthy and Cancerous Cells Dynamics in Acute Myeloid Leukemia
Authors: Abdelhafid Zenati, Mohamed Tadjine
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The mathematical formulation of biomedical problems is an important phase to understand and predict the dynamic of the controlled population. In this paper we perform a stability analysis of a coupled model for healthy and cancerous cells dynamics in Acute Myeloid Leukemia, this represents our first aim. Second, we illustrate the effect of the interconnection between healthy and cancer cells. The PDE-based model is transformed to a nonlinear distributed state space model (delay system). For an equilibrium point of interest, necessary and sufficient conditions of global asymptotic stability are given. Thus, we came up to give necessary and sufficient conditions of global asymptotic stability of the origin and the healthy situation and control of the dynamics of normal hematopoietic stem cells and cancerous during myelode Acute leukemia. Simulation studies are given to illustrate the developed results.Keywords: distributed delay, global stability, modelling, nonlinear models, PDE, state space
Procedia PDF Downloads 2527584 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
Procedia PDF Downloads 2767583 An Investigation of Anticancer Fluorinated Aza-Heterocycles
Authors: Darya O. Prima, Elena V. Vorontsova, Yuri G. Slizhov, Andrey V. Zibarev
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A broad family of carbocycle-fluorinated aza-heterocycles including 1,3-benzodiazoles (benzimidazoles), 1,2,3-benzotriazoles, 2,1,3-benzothia/selenadiazoles and 1,4-benzodiazines (quinoxalines) was synthesized in the unified way and assessed for cytotoxicity towards the Hep2 (laryngeal epidermoid carcinoma, a kind of oral cancer) cells. The diazoles, triazoles and selenadiazoles revealed low medium inhibitory concentrations IC50 = 2.2-26.4 µМ and induced the cells’ apoptosis at low concentrations C = 1-25 µМ. For selenadiazoles, cell death dynamics was observed already in the first hours after the treatment. Replacement of one atom F by group Me2N in some cases enlarged apoptotic activity of the compounds towards the Hep2 cells. In contrast, the archetypal (i.e. non-fluorinated) 1,3-benzodiazole, 1,2,3-benzotriazole and 2,1,3-benzoselenadiazole were low toxic (IC50 > 100 µM) and induced apoptosis only at high concentrations. The chlorinated congeners of the heterocycles under discussion were highly toxic towards the Hep2 cells but revealed insignificant ability to induce their apoptosis. Overall, the findings above suggest that fluorinated 1,3-benzodiazole, 1,2,3-benzotriazole and 2,1,3-benzoselenadiazole derivatives can be considered as potential anticancer drugs. For the laryngeal epidermoid carcinoma (for which, according to available statistics, the five-year survival rate remained ~50% during the past 30 years), it is especially important since surgical treatment is seriously complicated here thus encouraging medicament one.Keywords: Apoptosis, aza-heterocycles, cytotoxicity, fluorinated, Hep2 cells, synthesis
Procedia PDF Downloads 3397582 CRLH and SRR Based Microwave Filter Design Useful for Communication Applications
Authors: Subal Kar, Amitesh Kumar, A. Majumder, S. K. Ghosh, S. Saha, S. S. Sikdar, T. K. Saha
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CRLH (composite right/left-handed) based and SRR (split-ring resonator) based filters have been designed at microwave frequency which can provide better performance compared to conventional edge-coupled band-pass filter designed around the same frequency, 2.45 GHz. Both CRLH and SRR are unit cells used in metamaterial design. The primary aim of designing filters with such structures is to realize size reduction and also to realize novel filter performance. The CRLH based filter has been designed in microstrip transmission line, while the SRR based filter is designed with SRR loading in waveguide. The CRLH based filter designed at 2.45 GHz provides an insertion loss of 1.6 dB with harmonic suppression up to 10 GHz with 67 % size reduction when compared with a conventional edge-coupled band-pass filter designed around the same frequency. One dimensional (1-D) SRR matrix loaded in a waveguide shows the possibility of realizing a stop-band with sharp skirts in the pass-band while a stop-band in the pass-band of normal rectangular waveguide with tailoring of the dimensions of SRR unit cells. Such filters are expected to be very useful for communication systems at microwave frequency.Keywords: BPF, CRLH, harmonic, metamaterial, SRR and waveguide
Procedia PDF Downloads 4277581 Microfluidic Device for Real-Time Electrical Impedance Measurements of Biological Cells
Authors: Anil Koklu, Amin Mansoorifar, Ali Beskok
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Dielectric spectroscopy (DS) is a noninvasive, label free technique for a long term real-time measurements of the impedance spectra of biological cells. DS enables characterization of cellular dielectric properties such as membrane capacitance and cytoplasmic conductivity. We have developed a lab-on-a-chip device that uses an electro-activated microwells array for loading, DS measurements, and unloading of biological cells. We utilized from dielectrophoresis (DEP) to capture target cells inside the wells and release them after DS measurement. DEP is a label-free technique that exploits differences among dielectric properties of the particles. In detail, DEP is the motion of polarizable particles suspended in an ionic solution and subjected to a spatially non-uniform external electric field. To the best of our knowledge, this is the first microfluidic chip that combines DEP and DS to analyze biological cells using electro-activated wells. Device performance is tested using two different cell lines of prostate cancer cells (RV122, PC-3). Impedance measurements were conducted at 0.2 V in the 10 kHz to 40 MHz range with 6 s time resolution. An equivalent circuit model was developed to extract the cell membrane capacitance and cell cytoplasmic conductivity from the impedance spectra. We report the time course of the variations in dielectric properties of PC-3 and RV122 cells suspended in low conductivity medium (LCB), which enhances dielectrophoretic and impedance responses, and their response to sudden pH change from a pH of 7.3 to a pH of 5.8. It is shown that microfluidic chip allowed online measurements of dielectric properties of prostate cancer cells and the assessment of the cellular level variations under external stimuli such as different buffer conductivity and pH. Based on these data, we intend to deploy the current device for single cell measurements by fabricating separately addressable N × N electrode platforms. Such a device will allow time-dependent dielectric response measurements for individual cells with the ability of selectively releasing them using negative-DEP and pressure driven flow.Keywords: microfluidic, microfabrication, lab on a chip, AC electrokinetics, dielectric spectroscopy
Procedia PDF Downloads 1517580 Surface Modified Polyamidoamine Dendrimer with Gallic Acid Overcomes Drug Resistance in Colon Cancer Cells HCT-116
Authors: Khushbu Priyadarshi, Chandramani Pathak
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Cancer cells can develop resistance to conventional therapies especially chemotherapeutic drugs. Resistance to chemotherapy is another challenge in cancer therapeutics. Therefore, it is important to address this issue. Gallic acid (GA) is a natural plant compound that exhibits various biological properties including anti-proliferative, anti-inflammatory, anti-oxidant and anti-bacterial. Despite of the wide spectrum biological properties GA has cytotoxic response and low bioavailability. To overcome this problem, GA was conjugated with the Polyamidoamine(PAMAM) dendrimer for improving the bioavailability and efficient delivery in drug-resistant HCT-116 Colon Cancer cells. Gallic acid was covalently linked to 4.0 G PAMAM dendrimer. PAMAM dendrimer is well established nanocarrier but has cytotoxicity due to presence of amphiphilic nature of amino group. In our study we have modified surface of PAMAM dendrimer with Gallic acid and examine their anti-proliferative effects in drug-resistant HCT-116 cells. Further, drug-resistant colon cancer cells were established and thereafter treated with different concentration of PAMAM-GA to examine their anti-proliferative potential. Our results show that PAMAM-GA conjugate induces apoptotic cell death in HCT-116 and drug-resistant cells observed by Annexin-PI staining. In addition, it also shows that multidrug-resistant drug transporter P-gp protein expression was downregulated with increasing the concentration of GA conjugate. After that we also observed the significant difference in Rh123 efflux and accumulation in drug sensitive and drug-resistant cancer cells. Thus, our study suggests that conjugation of anti-cancer agents with PAMAM could improve drug resistant property and cytotoxic response to treatment of cancer.Keywords: drug resistance, gallic acid, PAMAM dendrimer, P-glycoprotein
Procedia PDF Downloads 1497579 The Conceptual and Procedural Knowledge of Rational Numbers in Primary School Teachers
Authors: R. M. Kashim
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The study investigates the conceptual and procedural knowledge of rational number in primary school teachers, specifically, the primary school teachers level of conceptual knowledge about rational number and the primary school teachers level of procedural knowledge about rational numbers. The study was carried out in Bauchi metropolis in Bauchi state of Nigeria. A Conceptual and Procedural Knowledge Test was used as the instrument for data collection, 54 mathematics teachers in Bauchi primary schools were involved in the study. The collections were analyzed using mean and standard deviation. The findings revealed that the primary school mathematics teachers in Bauchi metropolis posses a low level of conceptual knowledge of rational number and also possess a high level of Procedural knowledge of rational number. It is therefore recommended that to be effective, teachers teaching mathematics most posses a deep understanding of both conceptual and procedural knowledge. That way the most knowledgeable teachers in mathematics deliver highly effective rational number instructions. Teachers should not ignore the mathematical concept aspect of rational number teaching. This is because only the procedural aspect of Rational number is highlighted during instructions; this often leads to rote - learning of procedures without understanding the meanings. It is necessary for teachers to learn rational numbers teaching method that focus on both conceptual knowledge and procedural knowledge teaching.Keywords: conceptual knowledge, primary school teachers, procedural knowledge, rational numbers
Procedia PDF Downloads 3287578 The Effect of Manual Acupuncture-induced Injury as a Mechanism Contributing to Muscle Regeneration
Authors: Kamal Ameis
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This study aims to further improve our understanding of the underlying mechanism of local injury that occurs after manual acupuncture needle manipulation, and that initiates the muscle regeneration process, which is essential for muscle maintenance and adaptation. Skeletal muscle is maintained by resident stem cells called muscle satellite cells. These cells are normally in quiescent state, but following muscle injury, they re-enter the cell cycle and execute a myogenic program resulting in muscle fiber regeneration. Our previous work in young rats demonstrated that acupuncture treatment induced injury that activated resident satellite (stem) cells, which leads to muscle regeneration. Skeletal muscle regeneration is an adaptive response to injury that requires a tightly orchestrated event between signaling pathways activated by growth factor and intrinsic regulatory program controlled by myogenic transcription factor. We identified several gene expressions uniquely important for muscle regeneration in response to acupuncture treatment at different time course using different biological techniques, including Immunocytochemistry, western blotting, and Real Time PCR. This study uses a novel but non-invasive model of injury induced by manual acupuncture to further our current understanding of regenerative mechanism of muscle stem cells. From a clinical perspective, this model of injury induced by manual acupuncture may be easily translatable into a clinical tool that can be used as an alternative to physical exercise for patients challenged by bed rest or forced inactivity. Finally, the knowledge gained from this research could be useful for studies of the local effects of various modalities of induced injury, such as the traditional method of healing by cupping (hijamah), which may enhanced muscle stem cells and muscle fiber regeneration.Keywords: acupuncture, injury, regeneration, muscle stem cells
Procedia PDF Downloads 1487577 Computational Approaches to Study Lineage Plasticity in Human Pancreatic Ductal Adenocarcinoma
Authors: Almudena Espin Perez, Tyler Risom, Carl Pelz, Isabel English, Robert M. Angelo, Rosalie Sears, Andrew J. Gentles
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Pancreatic ductal adenocarcinoma (PDAC) is one of the most deadly malignancies. The role of the tumor microenvironment (TME) is gaining significant attention in cancer research. Despite ongoing efforts, the nature of the interactions between tumors, immune cells, and stromal cells remains poorly understood. The cell-intrinsic properties that govern cell lineage plasticity in PDAC and extrinsic influences of immune populations require technically challenging approaches due to the inherently heterogeneous nature of PDAC. Understanding the cell lineage plasticity of PDAC will improve the development of novel strategies that could be translated to the clinic. Members of the team have demonstrated that the acquisition of ductal to neuroendocrine lineage plasticity in PDAC confers therapeutic resistance and is a biomarker of poor outcomes in patients. Our approach combines computational methods for deconvolving bulk transcriptomic cancer data using CIBERSORTx and high-throughput single-cell imaging using Multiplexed Ion Beam Imaging (MIBI) to study lineage plasticity in PDAC and its relationship to the infiltrating immune system. The CIBERSORTx algorithm uses signature matrices from immune cells and stroma from sorted and single-cell data in order to 1) infer the fractions of different immune cell types and stromal cells in bulked gene expression data and 2) impute a representative transcriptome profile for each cell type. We studied a unique set of 300 genomically well-characterized primary PDAC samples with rich clinical annotation. We deconvolved the PDAC transcriptome profiles using CIBERSORTx, leveraging publicly available single-cell RNA-seq data from normal pancreatic tissue and PDAC to estimate cell type proportions in PDAC, and digitally reconstruct cell-specific transcriptional profiles from our study dataset. We built signature matrices and optimized by simulations and comparison to ground truth data. We identified cell-type-specific transcriptional programs that contribute to cancer cell lineage plasticity, especially in the ductal compartment. We also studied cell differentiation hierarchies using CytoTRACE and predict cell lineage trajectories for acinar and ductal cells that we believe are pinpointing relevant information on PDAC progression. Collaborators (Angelo lab, Stanford University) has led the development of the Multiplexed Ion Beam Imaging (MIBI) platform for spatial proteomics. We will use in the very near future MIBI from tissue microarray of 40 PDAC samples to understand the spatial relationship between cancer cell lineage plasticity and stromal cells focused on infiltrating immune cells, using the relevant markers of PDAC plasticity identified from the RNA-seq analysis.Keywords: deconvolution, imaging, microenvironment, PDAC
Procedia PDF Downloads 1287576 A Combination of Mesenchymal Stem Cells and Low-Intensity Ultrasound for Knee Meniscus Regeneration: A Preliminary Study
Authors: Mohammad Nasb, Muhammad Rehan, Chen Hong
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Background Meniscus defects critically alter knee function and lead to degenerative changes. Regenerative medicine applications including stem cell transplantation have showed a promising efficacy in finding alternatives to overcome traditional treatment limitations. However, stem cell therapy remains limited due to the substantially reduced viability and inhibitory microenvironment. Since tissue growth and repair are under the control of biochemical and mechanical signals, several approaches have recently been investigated (e.g., low intensity pulsed ultrasound [LIPUS]) to promote the regeneration process. This study employed LIPUS to improve growth and osteogenic differentiation of mesenchymal stem cells derived from human embryonic stem cells to improve the regeneration of meniscus tissue. Methodology: The Mesenchymal stromal cells (MSCs) were transplanted into the epicenter of the injured meniscus in rabbits, which were randomized into two main groups: a treatment group (n=32 New Zealand rabbits) including 4 subgroups of 8 rabbits in each subgroup (LIPUS treatment, MSC treatment, LIPUS with MSC and control), and a second group (n=9) to track implanted cells and their progeny using green fluorescence protein (GFP). GFP consists of the MSC and LIPUS-MSC combination subgroups. Rabbits were then subjected to histological, immunohistochemistry, and MRI assessment. Results: The quantity of the newly regenerated tissue in the combination treatment group that had Ultrasound irradiation after mesenchymal stem cells were better at all end points. Likewise, Tissue quality scores were also greater in knees treated with both approaches compared with controls and single treatment at all end points, achieving significance at twelve and twenty-four weeks [p < 0.05], and [p = 0.008] at twelve weeks. Differentiation into type-I and II collagen-expressing cells were higher in the combination group at up to twenty-four weeks. Conclusions: the combination of mesenchymal stem cells and LIPUS showed greater adhering to the sites of meniscus injury, differentiate into cells resembling meniscal fibrochondrocytes, and improve both quality and quantity of meniscal regeneration.Keywords: stem cells, regenerative medicine, osteoarthritis, knee
Procedia PDF Downloads 1197575 Trans-Activator of Transcription-Tagged Active AKT1 Variants for Delivery to Mammalian Cells
Authors: Tarana Siddika, Ilka U. Heinemann, Patrick O’Donoghue
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Protein kinase B (AKT1) is a serine/threonine kinase and central transducer of cell survival pathways. Typical approaches to study AKT1 biology in cells rely on growth factor or insulin stimulation that activates AKT1 via phosphorylation at two key regulatory sites (Threonine308, Serine473), yet cell stimulation also activates many other kinases and fails to differentiate the effect of the two main activating sites of AKT1 on downstream substrate phosphorylation and cell growth. While both AKT1 activating sites are associated with disease and used as clinical markers, in some cancers, high levels of Threonine308 phosphorylation are associated with poor prognosis while in others poor survival correlates with high Serine473 levels. To produce cells with specific AKT1 activity, a system was developed to deliver active AKT1 to human cells. AKT1 phospho-variants were produced from Escherichia coli with programmed phosphorylation by genetic code expansion. Tagging of AKT1 with an N-terminal cell penetrating peptide tag derived from the human immunodeficiency virus trans-activator of transcription (TAT) helped to enter AKT1 proteins in mammalian cells. The TAT-tag did not alter AKT1 kinase activity and was necessary and sufficient to rapidly deliver AKT1 protein variants that persisted in human cells for 24 h without the need to use transfection reagents. TAT-pAKT1T308, TAT-pAKT1S473 and TAT-pAKT1T308S473 proteins induced selective phosphorylation of the known AKT1 substrate GSK-3αβ, and downstream stimulation of the AKT1 pathway as evidenced by phosphorylation of ribosomal protein S6 at Serine240/244 in transfected cells. Increase in cell growth and proliferation was observed due to the transfection of different phosphorylated AKT1 protein variants compared to cells with TAT-AKT1 protein. The data demonstrate efficient delivery of AKT1 with programmed phosphorylation to human cells, thus establishing a cell-based model system to investigate signaling that is dependent on specific AKT1 activity and phosphorylation.Keywords: cell penetrating peptide, cell signaling, protein kinase b (AKT1), phosphorylation
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