Search results for: phenotype switch of smooth muscle cells
4494 On Lie Groupoids, Bundles, and Their Categories
Authors: P. G. Romeo
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A Lie group is a highly sophisticated structure which is a smooth manifold whose underlying set of elements is equipped with the structure of a group such that the group multiplication and inverse-assigning functions are smooth. This structure was introduced by the Norwegian mathematician So- phus Lie who founded the theory of continuous groups. The Lie groups are well developed and have wide applications in areas including Mathematical Physics. There are several advances and generalizations for Lie groups and Lie groupoids is one such which is termed as a "many-object generalization" of Lie groups. A groupoid is a category whose morphisms are all invertible, obviously, every group is a groupoid but not conversely. Definition 1. A Lie groupoid G ⇒ M is a groupoid G on a base M together with smooth structures on G and M such that the maps α, β: G → M are surjective submertions, the object inclusion map x '→ 1x, M → G is smooth, and the partial multiplication G ∗ G → G is smooth. A bundle is a triple (E, p, B) where E, B are topological spaces p: E → B is a map. Space B is called the base space and space E is called total space and map p is the projection of the bundle. For each b ∈ B, the space p−1(b) is called the fibre of the bundle over b ∈ B. Intuitively a bundle is regarded as a union of fibres p−1(b) for b ∈ B parametrized by B and ’glued together’ by the topology of the space E. A cross-section of a bundle (E, p, B) is a map s: B → E such that ps = 1B. Example 1. Given any space B, a product bundle over B with fibre F is (B × F, p, B) where p is the projection on the first factor. Definition 2. A principal bundle P (M, G, π) consists of a manifold P, a Lie group G, and a free right action of G on P denoted (u, g) '→ ug, such that the orbits of the action coincide with the fibres of the surjective submersion π : P → M, and such that M is covered by the domains of local sections σ: U → P, U ⊆ M, of π. Definition 3. A Lie group bundle, or LGB, is a smooth fibre bundle (K, q, M ) in which each fibre (Km = q−1(m), and the fibre type G, has a Lie group structure, and for which there is an atlas {ψi: Ui × G → KUi } such that each {ψi,m : G → Km}, is an isomorphism of Lie groups. A morphism of LGB from (K, q, M ) to (K′, q′, M′) is a morphism (F, f ) of fibre bundles such that each Fm: Km → K′ is a morphism of Lie groups. In this paper, we will be discussing the Lie groupoid bundles. Here it is seen that to a Lie groupoid Ω on base B there is associated a collection of principal bundles Ωx(B, Ωx), all of which are mutually isomorphic and conversely, associated to any principal bundle P (B, G, p) there is a groupoid called the Ehresmann groupoid which is easily seen to be Lie. Further, some interesting properties of the category of Lie groupoids and bundles will be explored.Keywords: groupoid, lie group, lie groupoid, bundle
Procedia PDF Downloads 774493 Nanomechanical Characterization of Healthy and Tumor Lung Tissues at Cell and Extracellular Matrix Level
Authors: Valeria Panzetta, Ida Musella, Sabato Fusco, Paolo Antonio Netti
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The study of the biophysics of living cells drew attention to the pivotal role of the cytoskeleton in many cell functions, such as mechanics, adhesion, proliferation, migration, differentiation and neoplastic transformation. In particular, during the complex process of malignant transformation and invasion cell cytoskeleton devolves from a rigid and organized structure to a more compliant state, which confers to the cancer cells a great ability to migrate and adapt to the extracellular environment. In order to better understand the malignant transformation process from a mechanical point of view, it is necessary to evaluate the direct crosstalk between the cells and their surrounding extracellular matrix (ECM) in a context which is close to in vivo conditions. In this study, human biopsy tissues of lung adenocarcinoma were analyzed in order to define their mechanical phenotype at cell and ECM level, by using particle tracking microrheology (PTM) technique. Polystyrene beads (500 nm) were introduced into the sample slice. The motion of beads was obtained by tracking their displacements across cell cytoskeleton and ECM structures and mean squared displacements (MSDs) were calculated from bead trajectories. It has been already demonstrated that the amplitude of MSD is inversely related to the mechanical properties of intracellular and extracellular microenvironment. For this reason, MSDs of particles introduced in cytoplasm and ECM of healthy and tumor tissues were compared. PTM analyses showed that cancerous transformation compromises mechanical integrity of cells and extracellular matrix. In particular, the MSD amplitudes in cells of adenocarcinoma were greater as compared to cells of normal tissues. The increased motion is probably associated to a less structured cytoskeleton and consequently to an increase of deformability of cells. Further, cancer transformation is also accompanied by extracellular matrix stiffening, as confirmed by the decrease of MSDs of matrix in tumor tissue, a process that promotes tumor proliferation and invasiveness, by activating typical oncogenic signaling pathways. In addition, a clear correlation between MSDs of cells and tumor grade was found. MSDs increase when tumor grade passes from 2 to 3, indicating that cells undergo to a trans-differentiation process during tumor progression. ECM stiffening is not dependent on tumor grade, but the tumor stage resulted to be strictly correlated with both cells and ECM mechanical properties. In fact, a greater stage is assigned to tumor spread to regional lymph nodes and characterized by an up-regulation of different ECM proteins, such as collagen I fibers. These results indicate that PTM can be used to get nanomechanical characterization at different scale levels in an interpretative and diagnostic context.Keywords: cytoskeleton, extracellular matrix, mechanical properties, particle tracking microrheology, tumor
Procedia PDF Downloads 2804492 The Effect of a Muscarinic Antagonist on the Lipase Activity
Authors: Zohreh Bayat, Dariush Minai-Tehrani
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Lipases constitute one of the most important groups of industrial enzymes that catalyze the hydrolysis of triacylglycerol to glycerol and fatty acids. Muscarinic antagonist relieves smooth muscle spasm of the gastrointestinal tract and effect on the cardiovascular system. In this research, the effect of a muscarinic antagonist on the lipase activity of Pseudomonas aeruginosa was studied. Lineweaver–Burk plot showed that the drug inhibited the enzyme by competitive inhibition. The IC50 value (60 uM) and Ki (30 uM) of the drug revealed the drug bound to the enzyme with high affinity. Determination of enzyme activity in various pH and temperature showed that the maximum activity of lipase was at pH 8 and 60°C both in presence and absence of the drug.Keywords: bacteria, inhibition, kinetics, lipase
Procedia PDF Downloads 4534491 The Effect of Blood Flow Restriction on the Knee Rehabilitation
Authors: O. Casasayas, M. Vigo, R. Navarro, P. Ragazzi, P. Alvarez, A. Perez-Bellmunt
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Introduction: The blood flow restriction training (BFR) is a method of muscle training that allows increasing the stress of muscle tissue to enhance the muscle cross-section and strength. This type of training has clear benefits in the rehabilitation field since it can improve muscle strength using low mechanical loads. The aim of this study is to know in which knee pathologies BFR has been used, what methodology was used and what were the obtained results. Study design: We performed a systematic literature search using strategies for the concepts of “blood flow restriction OR blood flow restriction training AND knee” in Medline. Articles were screened by authors and included if they used the blood flow restriction training in pathology of the knee. Results: The pathology more frequently treated by BFR was knee osteoarthritis and the variables most analyzed were strength and pain. The vascular occlusion used was 80% in the major part of studies. The groups of BFR obtained an increase of strength with less pain but not always the results are statistically significant. The evidence levels are poor in the high number of studies because in some cases there is not a control group or the evaluators were not blinded. Conclusion: The use of BFR is useful to improve muscle strength in knee pathology since it does not increase the pain, but more studies are needed to see (comprehend) if this type of treatment obtains better results than a conventional therapy. No studies have been found that compare the different occlusion effects in both the strength improvement and the pain reduction. Neither studies that analyse the effects of BFR on the muscle contractile parameters have been found.Keywords: blood flow restriction training, knee, arthroscopy knee, physical therapy
Procedia PDF Downloads 1684490 The Experience with SiC MOSFET and Buck Converter Snubber Design
Authors: Petr Vaculik
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The newest semiconductor devices on the market are MOSFET transistors based on the silicon carbide – SiC. This material has exclusive features thanks to which it becomes a better switch than Si – silicon semiconductor switch. There are some special features that need to be understood to enable the device’s use to its full potential. The advantages and differences of SiC MOSFETs in comparison with Si IGBT transistors have been described in first part of this article. Second part describes driver for SiC MOSFET transistor and last part of article represents SiC MOSFET in the application of buck converter (step-down) and design of simple RC snubber.Keywords: SiC, Si, MOSFET, IGBT, SBD, RC snubber
Procedia PDF Downloads 4834489 Activation of TNF-α from Human Endothelial Cells by Exposure of the Mitochondrial Stress Protein (Hsp60) Secreted from THP-1 Monocytes to High Glucose
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
Procedia PDF Downloads 1814488 Let-7 Mirnas Regulate Inflammatory Cytokine Production in Bovine Endometrial Cells after Lipopolysaccharide Challenge by Targeting TNFα
Authors: S. Ibrahim, D. Salilew-Wondim, M. Hoelker, C. Looft, E. Tholen, C. Grosse-Brinkhaus, K. Schellander, C. Neuhoff, D. Tesfaye
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Bovine endometrial cells appear to have a key role in innate immune defense of the female genital tract. A better understanding of molecular changes in microRNAs (miRNAs) and their target genes expression may identify reliable prognostic indicators for cows that will resolve inflammation and resume cyclicity. In the current study, we hypothesized that let-7 miRNAs family has a primary role in the innate immune defence of the endometrium tissue against bacterial infection, which is partly achieved via regulating mRNA stability of pro-inflammatory cytokines at the post-transcriptional level. Therefore, we conducted two experiments. In the first experiment, primary bovine endometrial cells were challenged with clinical (3.0 μg/ml) and sub-clinical (0.5 μg/ml) doses of lipopolysaccharide (LPS) for 24h. In the 2nd experiment, we have investigated the potential role of let-7 miRNAs (let-7a and let-7f) using gain and loss of function approaches. Additionally, tumor necrosis factor alpha (TNFα), transforming growth factor beta 1 induced transcript 1 (TGFB1I1) and serum deprivation response (SDPR) genes were validated using reporter assay. Here we addressed for the first time that let-7 miRNAs have a precise role in bovine endometrium, where LPS dysregulated let-7 miRNAs family expression was associated with an increased pro-inflammatory cytokine level by directly/indirectly targeting the TNFα, interleukin 6 (IL6), nuclear factor kappa-light-chain enhancer of activated B cells (NFκB), TGFβ1I1 and SDPR genes. To our knowledge, this is the first study showing that TNFα, TGFβ1I1 and SDPR were identified and validated as novel let-7 miRNAs targets and could have a distinct role in inflammatory immune response of LPS challenged bovine endometrial cells. Our data represent a new finding by which uterine homeostasis is maintained through functional regulation of let-7a by down-regulation of pro-inflammatory cytokines expression (TNFα and IL6) at the mRNA and protein levels. These findings suggest that LPS serves as a negative regulator of let-7 miRNAs expression and provides a mechanism for the persistent pro-inflammatory phenotype, which is a hallmark of bovine subclinical endometritis.Keywords: bovine endometrial cells, let-7, lipopolysaccharide, pro-inflammatory cytokines
Procedia PDF Downloads 3804487 Effectiveness of Impairment Specified Muscle Strengthening Programme in a Group of Disabled Athletes
Authors: A. L. I. Prasanna, E. Liyanage, S. A. Rajaratne, K. P. A. P. Kariyawasam, A. A. J. Rajaratne
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Maintaining or improving the muscle strength of the injured body part is essential to optimize performance among disabled athletes. General conditioning and strengthening exercises might be ineffective if not sufficiently intense enough or targeted for each participant’s specific impairment. Specific strengthening programme, targeted to the affected body part, are essential to improve the strength of impaired muscles and increase in strength will help reducing the impact of disability. Methods: The muscle strength of hip, knee and ankle joints was assessed in a group of randomly selected disabled athletes, using the Medical Research Council (MRC) grading. Those having muscle strength of grade 4 or less were selected for this study (24 in number) and were given and a custom made exercise program designed to strengthen their hip, knee or ankle joint musculature, according to the muscle or group of muscles affected. Effectiveness of the strengthening program was assessed after a period of 3 months. Results: Statistical analysis was done using the Minitab 16 statistical software. A Mann-Whitney U test was used to compare the strength of muscle group before and after exercise programme. A significant difference was observed after the three month strengthening program for knee flexors (Left and Right) (P =0.0889, 0.0312) hip flexors (left and right) (P=0.0312, 0.0466), hip extensors (Left and Right) (P=0.0478, 0.0513), ankle plantar flexors (Left and Right) (P=0.0466, 0.0423) and right ankle dorsiflexors (P= 0.0337). No significant difference of strength was observed after the strengthening program in the knee extensors (left and right), hip abductors (left and right) and left ankle dorsiflexors. Conclusion: Impairment specific exercise programme appear to be beneficial for disabled athletes to significantly improve the muscle strength of the affected joints.Keywords: muscle strengthening programme, disabled athletes, physiotherapy, rehabilitation sciences
Procedia PDF Downloads 3574486 Development of a Natural Anti-cancer Formulation Which Can Target Triple Negative Breast Cancer Stem Cells
Authors: Samashi Munaweera
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Cancer stem cells (CSC) are responsible for the initiation, extensive proliferation and metastasis of cancer. CSCs, including breast cancer stem cells (bCSCs) have a capacity to generate chemo and radiotherapy resistance heterogeneous population of cells. Over-expressed ABCB1 has been reported as a main reason for drug resistance of CSCs via activating drug efflux pumps by creating pores in the cell membrane. The overall efficiency of chemotherapeutic agents might be enhanced by blocking the ABCB protein efflux pump in the CSC membrane. There is an urgent need to search for persuasive natural drugs which can target CSCs. Anti-cancer properties of Hylocereus undatus on cancer CSCs have not yet been studied. In the present study, the anti-cancer effects of the peel and flesh of H. undatus fruit on bCSCs were evaluated with the aim of developing a marketable anti-cancer nutraceutical formulation. The flesh and peel of H. undatus were freeze-dried and sequentially extracted into four different solvents (hexane, chloroform, ethyl acetate and ethanol). All extracts (eight extracts) were dried under reduced pressure, and different concentrations (12.5-400 µg/mL) were treated on bCSCs isolated from a triple-negative chemo-resistant breast cancer phenotype (MDA-MB-231 cells). Anti-proliferative effects of all extracts and paclitaxel (positive control) were determined by a colorimetric assay (WST-1 based). Since peel-chloroform (IC50= 54.8 µg/mL) and flesh-ethyl acetate (IC50= 150.5 µg/mL) extras exerted a potent anti-proliferative effect at 72 h post-incubation, a combinatorial formulation (CF) was developed with the most active peel-chloroform extract and 20 µg/mL of verapamil (a known ABCB1 drug efflux pump blocker) first time in the world. Anti-proliferative effects and pro-apoptotic effects of CF were confirmed by estimating activated caspase3 and caspase7 levels and apoptotic morphological features in the CF-treated bCSCs compared to untreated and only verapamil (20 µg/mL) treated bCSCs, and CF treated normal mammary epithelial cells (MCF-10A). The antiproliferative effects of CF (16.4 µg/mL) are greater than paclitaxel (19.2 µg/mL) and three folds greater than peel-chloroform extract (IC50= 54.8 µg/mL) on bCSCs while exerting less effects on normal cells (> 400 µg/mL). Collectively, CF can be considered as a potential initiative of a nutraceutical formulation that can target CSCs.Keywords: breast cancer stem cells (bCSCs), Hylocereus undatus, combinatorial formulation (CF), ABCB 1 protein, verapamil
Procedia PDF Downloads 274485 Designing Floor Planning in 2D and 3D with an Efficient Topological Structure
Authors: V. Nagammai
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Very-large-scale integration (VLSI) is the process of creating an integrated circuit (IC) by combining thousands of transistors into a single chip. Development of technology increases the complexity in IC manufacturing which may vary the power consumption, increase the size and latency period. Topology defines a number of connections between network. In this project, NoC topology is generated using atlas tool which will increase performance in turn determination of constraints are effective. The routing is performed by XY routing algorithm and wormhole flow control. In NoC topology generation, the value of power, area and latency are predetermined. In previous work, placement, routing and shortest path evaluation is performed using an algorithm called floor planning with cluster reconstruction and path allocation algorithm (FCRPA) with the account of 4 3x3 switch, 6 4x4 switch, and 2 5x5 switches. The usage of the 4x4 and 5x5 switch will increase the power consumption and area of the block. In order to avoid the problem, this paper has used one 8x8 switch and 4 3x3 switches. This paper uses IPRCA which of 3 steps they are placement, clustering, and shortest path evaluation. The placement is performed using min – cut placement and clustering are performed using an algorithm called cluster generation. The shortest path is evaluated using an algorithm called Dijkstra's algorithm. The power consumption of each block is determined. The experimental result shows that the area, power, and wire length improved simultaneously.Keywords: application specific noc, b* tree representation, floor planning, t tree representation
Procedia PDF Downloads 3934484 Effects of Butea superba Roxb. on Skeletal Muscle Functions and Parvalbumin Levels of Orchidectomized Rat
Authors: Surapong Vongvatcharanon, Fardeela Binalee, Wandee Udomuksorn, Ekkasit Kumarnsit, Uraporn Vongvatcharanon
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Hypogonadism is characterized by a decline in sex hormone levels, especially testosterone. It has been shown to be an important contributor to the decrease in muscle mass, muscle strength and performance, a condition known as sarcopenia. Preparations from Butea superba Roxb. (red Kwao Krua) have been reported to have androgenic properties. The active compounds are proposed to be flavonoids and flavonoid glycosides. Treatment with B. superba has been shown to improve erectile dysfunction in males. Parvalbumin (PV) is a relaxing factor and identified in fast twitch fibers. Alterations of the PV levels affects skeletal muscle functions. This study aimed to investigate the effects of orhchidectomy, testosterone replacement and different doses of Butea superba Roxb. on the structure, performance, levels of parvalbumin, parvalbumin and androgen receptor immunoreactivities in the extensor digitorum longus (EDL) and gastrocnemius muscles of orchidectomized rats. Twelve-week old male Wistar rats were randomly divided into 6 groups; sham-operated (SHAM), orchidectomized (BS-0), orchidectomized group that was treated with testosterone replacement of 6 µg/kg (TP) or an orchidectomized group that was treated with various doses of an extract from Butea superba Roxb.; 5 mg/kg (BS-5), 50 mg/kg (BS-50) and 500 mg/kg (BS-500) all for 90 days. The testosterone level, epididymis, seminal vesicle, prostate gland, vas deference weight, muscle fiber size, strength and endurance in both the EDL and gastrocnemius muscle were decreased in the BS-0 group but increased in the testosterone replacement group. Treatment with the B. superba Roxb. extract replacement group improved muscle fiber size, strength and endurance, but not total testosterone levels, or the epididymis, seminal vesicle, prostate gland, vas deference weight. Furthermore, the parvalbumin level, parvalbumin and androgen receptor immunoreactivities were reduced in the BS-0 group but increased in the testosterone replacement group and the B. superba Roxb. extract groups for both the EDL and gastrocnemius muscle. This study indicated that the reduction of testosterone level led to a decrease of the androgen receptor density resulting in a decline in the muscle mass and parvalbumin levels. The decrease of parvalbumin levels affected muscle performance. Testosterone replacement increased the androgen receptor density and led to an increase of muscle mass and parvalbumin levels. The increase in the parvalbumin levels may result in an improvement of muscle performance. This may explain one mechanism of testosterone on muscle mass and strength in the testosterone dependent sarcopenia. The B. superba Roxb. extract groups also had improved muscle mass, strength and endurance, parvalbumin level, parvalbumin and androgen immunoreactivities compared to the BS-O group . Butea superba Roxb. Extracts contains a flavonoid (3, 7, 3'-Trihydroxy-4'-methoxyflavone), flavonoiglycoside (3, 3'-dihydroxy-4'-methoxyflavone-7-O-β-D-glucopyranoside) and isoflavanolignans (butesuperins A and butesuperins B) all known to inhibit the cAMP phosphodiesterase enzyme. Therefore, cAMP signaling may have adaptive effects on skeletal muscle by increasing muscle mass, strength and endurance.Keywords: Butea superba, parvalbumin, skeletal muscle, orchidectomy
Procedia PDF Downloads 4234483 Appropriate Depth of Needle Insertion during Rhomboid Major Trigger Point Block
Authors: Seongho Jang
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Objective: To investigate an appropriate depth of needle insertion during trigger point injection into the rhomboid major muscle. Methods: Sixty-two patients who visited our department with shoulder or upper back pain participated in this study. The distance between the skin and the rhomboid major muscle (SM) and the distance between the skin and rib (SB) were measured using ultrasonography. The subjects were divided into 3 groups according to BMI: BMI less than 23 kg/m2 (underweight or normal group); 23 kg/m2 or more to less than 25 kg/m2 (overweight group); and 25 kg/m2 or more (obese group). The mean ±standard deviation (SD) of SM and SB of each group were calculated. A range between mean+1 SD of SM and the mean-1 SD of SB was defined as a safe margin. Results: The underweight or normal group’s SM, SB, and the safe margin were 1.2±0.2, 2.1±0.4, and 1.4 to 1.7 cm, respectively. The overweight group’s SM and SB were 1.4±0.2 and 2.4±0.9 cm, respectively. The safe margin could not be calculated for this group. The obese group’s SM, SB, and the safe margin were 1.8±0.3, 2.7±0.5, and 2.1 to 2.2 cm, respectively. Conclusion: This study will help us to set the standard depth of safe needle insertion into the rhomboid major muscle in an effective manner without causing any complications.Keywords: pneumothorax, rhomboid major muscle, trigger point injection, ultrasound
Procedia PDF Downloads 2904482 Regulated Output Voltage Double Switch Buck-Boost Converter for Photovoltaic Energy Application
Authors: M. Kaouane, A. Boukhelifa, A. Cheriti
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In this paper, a new Buck-Boost DC-DC converter is designed and simulated for photovoltaic energy system. The presented Buck-Boost converter has a double switch. Moreover, its output voltage is regulated to a constant value whatever its input is. In the presented work, the Buck-Boost transfers the produced energy from the photovoltaic generator to an R-L load. The converter is controlled by the pulse width modulation technique in a way to have a suitable output voltage, in the other hand, to carry the generator’s power, and put it close to the maximum possible power that can be generated by introducing the right duty cycle of the pulse width modulation signals that control the switches of the converter; each component and each parameter of the proposed circuit is well calculated using the equations that describe each operating mode of the converter. The proposed configuration of Buck-Boost converter has been simulated in Matlab/Simulink environment; the simulation results show that it is a good choice to take in order to maintain the output voltage constant while ensuring a good energy transfer.Keywords: Buck-Boost converter, switch, photovoltaic, PWM, power, energy transfer
Procedia PDF Downloads 9054481 Characterization of a Mesenchymal Stem Cells Pool in Killian Nasal Polyp
Authors: Emanuela Chiarella, Clelia Nisticò, Nicola Lombardo, Giovanna Lucia Piazzetta, Nadia Lobello, Maria Mesuraca
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Killian’s Antrochoanal Polyp is a benign lesion of the maxillary sinus characterized by unilateral nasal obstruction, pus discharge, and headache. It affects, more commonly children and young adults. Although its etiology still remains unclear, chronic inflammation, autoreactivity, allergies, and viral infections are strongly associated with its formation and development, resulting in nasal tissue remodeling. We aimed to investigate the stem cells components which reside in this pathological tissue. In particular, we adopted a protocol for the isolation and culturing of mesenchymal stem cells from surgical biopsies of three Killian nasal polyp patients (KNP-MSCs) as well as from their healthy nasal tissue (HNT-MSCs) that were used as controls. The immunophenotype profile of HNT-MSCs and KNP-MSCs was more similar, with a marked positivity for CD73, CD90, and CD105 expression, while being negative for CD34 and CD14 haematopoietic genes. Cell proliferation assay showed that KNP-MSCs had a replicative disadvantage compared to HNT-MSCs, as evidenced by the significantly lower number of cells in the S-phase of the cell cycle. KNP-MSCs also took longer to close a wound than HNT-MSCs, indicating a partial epithelial phenotype in which low levels of ICAM-1 mRNA and a significant increase in E-CAD transcript were detectable. Subsequently, the differentiation potential of both MSCs populations was analyzed by inducing osteoblastic or adipocyte differentiation for up to 20 days. KNP-MSCs showed the ability to differentiate into osteoblasts, although ALP activity as well as the number and size of calcium deposits were lower than osteogenic induced-HNT-MSCs. Also, mRNA levels of osteoblastic marker genes (OCN, OPN, OSX, RUNX2) resulted lower compared to control cell population. Instead, the analysis of the adipogenic differentiation potential showed a similar behavior between KNP-MSCs and HNT-MSCs considering that the amount of lipid droplets, the expression of adipocyte-specific genes (FABP4, AdipoQ, PPARγ2, LPL) and the content of triacylglycerols were almost overlapping. Taken together, these results first demonstrated that Killian's nasal polyp is a source of mesenchymal stem cells with self-renewal and multi-differentiative capabilities.Keywords: Mesenchymal stem cells, adipogenic differentiation, osteogenic differentiation, EMT
Procedia PDF Downloads 774480 Decreased Autophagy Contributes to Senescence Induction in HS68 Cells
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
Procedia PDF Downloads 2554479 90-Day Strength Training Intervention Decreases Incidence of Sarcopenia: A Pre- and Posttest Pilot Study of Older Adults in a Skilled Nursing Facility
Authors: Donna-Marie Phyllis Lanton
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Sarcopenia is a well-known geriatric syndrome characterized by the progressive and generalized loss of muscle quantity or quality. The incidence of sarcopenia increases with age and is associated with adverse outcomes such as the increased risk of falls, cognitive impairment, loss of independence, diminished quality of life, increased health costs, need for care in a skilled nursing facility, and increased mortality. Physical activity, including resistance training, is the most prevalent recommendation for treating and preventing sarcopenia. Residents (N = 23) of a skilled nursing facility in East Orlando, Florida, participated in a 90-day strength training program designed using the PARIHS framework to improve measures of muscle mass, muscle strength, physical performance, and quality of life. Residents engaged in both resistance and balance exercises for 1 hour two times a week. Baseline data were collected and compared to data at Days 30, 60, and 90. T tests indicated significant gains on all measures from baseline to 90 days: muscle mass increased by 1.2 (t[22] = 2.85, p = .009), grip strength increased by 4.02 (t[22] = 8.15, p < .001), balance increased by 2.13 (t[22] = 18.64, p < .001), gait speed increased by 1.83 (t[22] = 17.84, p < .001), chair speed increased 1.87 (t[22] = 16.36, p < .001), and quality of life score increased by 17.5 (t[22] = 19.26, p < .001). For residents with sarcopenia in skilled nursing facilities, a 90-day strength training program with resistance and balance exercises may provide an option for decreasing the incidence of sarcopenia among that populationKeywords: muscle mass, muscle strength, older adults, PARIHS framework
Procedia PDF Downloads 884478 Defective Autophagy Leads to the Resistance to PP2 in ATG5 Knockout Cells Generated by CRISPR-Cas9 Endonuclease
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
Procedia PDF Downloads 3474477 Effect of Oxytocin on Cytosolic Calcium Concentration of Alpha and Beta Cells in Pancreas
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
Procedia PDF Downloads 1924476 The Effects of Dietary Flaxseed Oil Supplementations on Punicic Acid of the Yolks in Quail Muscle
Authors: Ozcan Baris Citil
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This experiment was carried out to determine effects of Japanese quail diets which is supplemented with different levels of punicic acids and CLA compositions in Japanese quail. In this study, eighty laying quails at 12 weeks of age were used. They were divided into 4 tretament groups, each group included 20 quails. The diets in treatment groups contained different levels of flaxseed oil. At the end of the experiment lasted for 21 days, 5 quail taken from each group were subjected to analysis. Punicic acid content of muscle were determined by gas chromatography. Twenty five different fatty acid components were determinated in the compositions of quail muscle. No differences were found in oil content among the groups.Keywords: quail egg yolk, punicic acid, flaxseed oil, gas chromatography
Procedia PDF Downloads 2924475 An Electromyographic Study of Muscle Coordination during Dynamic Glenohumeral Joint Elevation
Authors: Omid Khaiyat, David Hawkes
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Introduction: There remains a lack of information on sophisticated coordination patterns across shoulder girdle muscles. Considering the stability of the shoulder being heavily dependent on coordinated muscle activity during its wide-ranging movements, it is important that key intermuscular relationships are well-defined for a better understanding of underlying pathology. This study investigated shoulder intermuscular coordination during different planes of shoulder elevation. Materials and Methods: EMG was recorded from 14 shoulder muscles in 20 healthy participants during shoulder flexion, scapula plane elevation, abduction, and extension. Cross-correlation by means of Pearson Correlation Coefficient (PCC) was used to examine the coordination between different muscles and muscle groups. Results: Coordination between rotator cuff and deltoid muscle groups was significantly higher (p =0.020-0.035) during the initial (PCC) = 0.79) and final (PCC = 0.74) phases of elevation compared to the mid-range (PCC = 0.34). Furthermore, a high level of coordination (PCC = 0.89) was noted between the deltoid group and the adductor group (latissimus dorsi and teres major) during the initial stage of shoulder elevation. Conclusion: The destabilising force of the deltoid during the initial stage of shoulder elevation is balanced by coordinated activity of rotator cuff, latissimus dorsi, and teres major. This is also the case for the end-range of movement, where increased demand for stability again leads to higher coordination between the deltoid and rotator cuff muscle groups. Appreciation of the sophistication of normal shoulder function evidence-based rehabilitation strategies for conditions such as subacromial impingement syndrome or shoulder instability can be developed.Keywords: shoulder, coordination, EMG, muscle activity, upper limb
Procedia PDF Downloads 1664474 The Conjugated Polymers in improving the Organic Solar Cells Efficiency
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
Procedia PDF Downloads 854473 Mechanical Responses to Hip Versus Knee Induced Muscle Fatigue in Patellofemoral Pain Syndrome
Authors: Eman Ahmed Ahmed, Ghada Abdelmoneim Mohamed, Hamada Ahmed Hamada, Nagui Sobhi Nassif
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Impaired skeletal muscle endurance may be an important causal factor in the development of patellofemoral pain syndrome (PFPS). However, there is lack of information regarding the effect of hip versus knee muscle fatigue on isokinetic parameters, and myoelectric activity of hip and knee muscles in these patients. Purpose: The study was conducted to investigate the effect of hip abductors versus knee extensors fatigue protocol on knee proprioception, hip and knee muscle strength and their myoelectric activity in patients with PFPS. Methods: Fifteen female patients with PFPS participated in the study. They were tested randomly under two fatiguing conditions; hip abductors and knee extensors fatigue protocols. Isolated muscle fatigue of two muscles was induced isokinetically on the affected side in a two separate sessions with a rest interval of at least three days. After determining peak torque, patients performed continuous maximal concentric-eccentric contraction of the selected muscle until the torque output dropped below 50% of peak torque value for 3 consecutive repetitions. Knee proprioception, eccentric hip abductors' peak torque, eccentric knee extensors' peak torque, EMG ratio of vastus medialis obliquus (VMO) / vastus lateralis (VL), and EMG activity of gluteus medius (GM) muscle, were recorded before and immediately after each fatigue protocol using the Biodex Isokinetic system and EMG Myosystem. Results: Two-way within subject MANOVA revealed that eccentric knee extensors’ peak torque decreased significantly after hip abductors fatigue protocol compared to pre fatigue condition (p<0.05). On the other hand, there was no statistically significant difference in the eccentric hip abductors’ peak torque after admitting knee extensors fatigue protocol (p > 0.05). Moreover, no significant difference was found in knee proprioception, EMG ratio of VMO/VL, and EMG activity of GM muscle, after either hip or knee fatigue protocol (p>0.05). Conclusion: A hip focused rehabilitation program may be beneficial in improving knee function through correcting faulty kinematics and hence decrease knee loading in patients with PFPS.Keywords: electromyography, knee proprioception, mechanical responses, muscle fatigue, patellofemoral pain syndrome
Procedia PDF Downloads 3114472 Effect of Different Muscle Contraction Mode on the Expression of Myostatin, IGF-1, and PGC-1 Alpha Family Members in Human Vastus Lateralis Muscle
Authors: Pejman Taghibeikzadehbadr
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Muscle contraction stimulates a transient change of myogenic factors, partly related to the mode of contractions. Here, we assessed the response of Insulin-like growth factor 1Ea (IGF-1Ea), Insulin-like growth factor 1Eb (IGF-1Eb), Insulin-like growth factor 1Ec (IGF-1Ec), Peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC1α-1), Peroxisome proliferator-activated receptor gamma coactivator 4-alpha (PGC1α-4), and myostatin to the eccentric Vs the concentric contraction in human skeletal muscle. Ten healthy males were performed an acute eccentric and concentric exercise bout (n = 5 per group). For each contraction type, participants performed 12 sets of 10 repetitions knee extension by the dominant leg. Baseline and post-exercise muscle biopsy were taken 4 weeks before and immediately after experimental sessions from Vastus Lateralis muscle. Genes expression was measured by real-time PCR technique. There was a significant increase in PGC1α-1, PGC1α-4, IGF-1Ea and, IGF-1Eb mRNA after concentric contraction (p ≤ 0.05), while the PGC1α-4 and IGF-1Ec significantly increased after eccentric contraction (p ≤ 0.05). It is intriguing to highlight that; no significant differences between groups were evident for changes in any variables following exercise bouts (p ≥ 0.05). Our results found that concentric and eccentric contractions presented different responses in PGC1α-1, IGF-1Ea, IGF-1Eb, and IGF-1Ec mRNA. However, a similar significant increase in mRNA content was observed in PGC1α-4. Further, no apparent differences could be found between the response of genes to eccentric and concentric contraction.Keywords: eccentric contraction, concentric contraction, gene expression, PGC-1 alpha, IGF-1 Myostatin
Procedia PDF Downloads 1604471 MicroRNA-211 Regulates Oxidative Phosphorylation and Energy Metabolism in Human Vitiligoa
Authors: Anupama Sahoo, Bongyong Lee, Katia Boniface, Julien Seneschal, Sanjaya K. Sahoo, Tatsuya Seki, Chunyan Wang, Soumen Das, Xianlin Han, Michael Steppie, Sudipta Seal, Alain Taieb, Ranjan J. Perera
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Vitiligo is a common, chronic skin disorder characterized by loss of epidermal melanocytes and progressive depigmentation. Vitiligo has a complex immune, genetic, environmental, and biochemical etiology, but the exact molecular mechanisms of vitiligo development and progression, particularly those related to metabolic control, are poorly understood. Here we characterized the human vitiligo cell line PIG3V and the normal human melanocytes, HEM-l by RNA-sequencing, targeted metabolomics, and shotgun lipidomics. Melanocyte-enriched miR-211, a known metabolic switch in non-pigmented melanoma cells, was severely downregulated in vitiligo cell line PIG3V and skin biopsies from vitiligo patients, while its novel predicted targets transcriptional co-activator PGC1-α (PPARGC1A), ribonucleotide reductase regulatory subunit M2 (RRM2), and serine-threonine protein kinase TAO1 (TAOK1) were reciprocally upregulated. miR-211 binds to PGC1-α 3’UTR locus and represses it. Although mitochondrial numbers were constant, mitochondrial complexes I, II, and IV and respiratory responses were defective in vitiligo cells. Nanoparticle-coated miR-211 partially augmented the oxygen consumption rate in PIG3V cells. The lower oxygen consumption rate, changes in lipid and metabolite profiles, and increased reactive oxygen species production observed in vitiligo cells appear to be partly due to abnormal regulation of miR-211 and its target genes. These genes represent potential biomarkers and therapeutic targets in human vitiligo.Keywords: metabolism, microRNA, mitochondria, vitiligo
Procedia PDF Downloads 3674470 A Self-Adaptive Stimulus Artifacts Removal Approach for Electrical Stimulation Based Muscle Rehabilitation
Authors: Yinjun Tu, Qiang Fang, Glenn I. Matthews, Shuenn-Yuh Lee
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This paper reports an efficient and rigorous self-adaptive stimulus artifacts removal approach for a mixed surface EMG (Electromyography) and stimulus signal during muscle stimulation. The recording of EMG and the stimulation of muscles were performing simultaneously. It is difficult to generate muscle fatigue feature from the mixed signal, which can be further used in closed loop system. A self-adaptive method is proposed in this paper, the stimulation frequency was calculated and verified firstly. Then, a mask was created based on this stimulation frequency to remove the undesired stimulus. 20 EMG signal recordings were analyzed, and the ANOVA (analysis of variance) approach illustrated that the decreasing trend of median power frequencies was successfully generated from the 'cleaned' EMG signal.Keywords: EMG, FES, stimulus artefacts, self-adaptive
Procedia PDF Downloads 3994469 Symmetry of Performance across Lower Limb Tests between the Dominant and Non-Dominant Legs
Authors: Ghulam Hussain, Herrington Lee, Comfort Paul, Jones Richard
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Background: To determine the functional limitations of the lower limbs or readiness to return to sport, most rehabilitation programs use some form of testing; however, it is still unknown what the pass criteria is. This study aims to investigate the differences between the dominant and non-dominant leg performances across several lower limb tasks, which are hop tests, two-dimensional (2D) frontal plane projection angle (FPPA) tests, and isokinetic muscle tests. This study also provides the reference values for the limb symmetry index (LSI) for the hop and isokinetic muscle strength tests. Twenty recreationally active participants were recruited, 11 males and 9 females (age 23.65±2.79 years; height 169.9±3.74 cm; and body mass 74.72±5.81 kg. All tests were undertaken on the dominant and non-dominant legs. These tests are (1) Hop tests, which include horizontal hop for distance and crossover hop tests, (2) Frontal plane projection angle (FPPA): 2D capturing from two different tasks, which are forward hop landing and squatting, and (3) Isokinetic muscle strength tests: four different muscles were tested: quadriceps, hamstring, ankle plantar flexor, and hip extensor muscles. The main outcome measurements were, for the (1) hop tests: maximum distance was taken when undertaking single/crossover hop for distance using a standard tape measure, (2) for the FPPA: the knee valgus angle was measured from the maximum knee flexion position using a single 2D camera, and (3) for the isokinetic muscle strength tests: three different variables were measured: peak torque, peak torque to body weight, and the total work to body weight. All the muscle strength tests have been applied in both concentric and eccentric muscle actions at a speed of 60°/sec. This study revealed no differences between the dominant and non-dominant leg performance, and 85% of LSI was achieved by the majority of the subjects in both hop and isokinetic muscle tests, and; therefore, one leg’s hop performance can define the other.Keywords: 2D FPPA, hop tests, isokinetic testing, LSI
Procedia PDF Downloads 664468 Investigation of the Excitotoxicity Pathways in Neuroblastoma Cells
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
Procedia PDF Downloads 1584467 Up-Regulation of SCUBE2 Expression in Co-Cultures of Human Mesenchymal Stem Cell and Breast Cancer Cells
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
Procedia PDF Downloads 3404466 The Effect of Low Power Laser on CK and Some of Markers Delayed Onset Muscle Soreness (DOMS)
Authors: Bahareh Yazdanparast Chaharmahali
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The study showed effect of low power laser therapy on knee range of motion (flexion and extension), resting angle of knee joint, knee circumference and rating of delayed onset muscle soreness induced pain, 24 and 48 hours after eccentric training of knee flexor muscle (hamstring muscle). We investigate the effects of pulsed ultrasound on swelling, relaxed, flexion and extension knee angle and pain. 20 volunteers among girl students of college voluntary participated in this research. After eccentric training, subjects were randomly divided into two groups, control and laser therapy. In day 1 and in order to induce delayed onset muscle soreness, subjects eccentrically trained their knee flexor muscles. In day 2, subjects were randomly divided into two groups: control and low power laser therapy. 24 and 48 hours after eccentric training. Variables (knee flexion and extension, srang of motion, resting knee joint angle and knee circumferences) were measured and analyzed. Data are reported as means ± standard error (SE) and repeated measured was used to assess differences within groups. Methods of treatment (low power laser therapy) have significant effects on delayed onset muscle soreness markers. 24 and 48 hours after training a significant difference was observed between mean pains of 2 groups. This difference was significant between low power laser therapy and C groups. The Bonferroni post hock is significant. Low power laser therapy trophy as used in this study did significantly diminish the effects of delayed – onset muscle soreness on swelling, relaxed – knee extension and flexion angle.Keywords: creatine kinase, DOMS, eccentric training, low power laser
Procedia PDF Downloads 2464465 The Effect of Progressive Muscle Relaxation and Sleep Hygiene Education to Change Sleep Quality Index Scores of Patient with Breast Cancer
Authors: Ika Wulansari, Yati Afiyanti, Indang Trihandini
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Sleeping disorder experienced by patients with breast cancer can affect the physical, mental, health, and well-being. This study examines the effect of progressive muscle relaxation training and sleep hygiene education to change sleep quality scores of the patient with breast cancer. The study design using quasi-experiment with pre-post test within the control group, involving 62 breast cancer patients using consecutive sampling method in Jakarta. Statistical test results with independent t-test showed a significant difference in score of sleep quality between in intervention group and the control group (6,66±3,815; 9,30±3,334, p-value = 0,005). Progressive muscle relaxation exercise and sleep hygiene education proven to be affective to change the patients sleeping quality, so that it can be an alternative therapeutic option to overcome sleeping disorders.Keywords: sleeping disorders, breast cancer, progressive muscle relaxation, sleep hygiene education
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