Search results for: skeletal muscle cells C2C12
Commenced in January 2007
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Edition: International
Paper Count: 3840

Search results for: skeletal muscle cells C2C12

3570 The Correlation between Body Composition and Spinal Alignment in Healthy Young Adults

Authors: Ferruh Taspinar, Ismail Saracoglu, Emrah Afsar, Eda O. Okur, Gulce K. Seyyar, Gamze Kurt, Betul Taspinar

Abstract:

Although it is thought that abdominal adiposity is one of the risk factor for postural deviation, such as increased lumbar lordosis, the body mass index is not sufficient to indicate effects of abdominal adiposity on spinal alignment and postural changes. The aim of this study was to investigate the correlation with detailed body composition and spine alignment in healthy young adults. This cross-sectional study was conducted with sixty seven healthy volunteers (37 men and 30 women) whose ages ranged between 19 and 27 years. All participants’ sagittal spinal curvatures of lumbar and thoracic region were measured via Spinal mouse® (Idiag, Fehraltorf, Switzerland). Also, body composition analysis (whole body fat ratio, whole body muscle ratio, abdominal fat ratio, and trunk muscle ratio) estimation by means of bioelectrical impedance was evaluated via Tanita Bc 418 Ma Segmental Body Composition Analyser (Tanita, Japan). Pearson’s correlation was used to analysis among the variables. The mean lumbar lordosis and thoracic kyphosis angles were 21.02°±9.39, 41.50°±7.97, respectively. Statistically analysis showed a significant positive correlation between whole body fat ratio and lumbar lordosis angle (r=0.28, p=0.02). Similarly, there was a positive correlation between abdominal fat ratio and lumbar lordosis angle (r=0.27, p=0.03). The thoracic kyphosis angle showed also positive correlation with whole body fat ratio (r=0.33, p=0.00) and abdominal fat ratio (r=0.40, p=0.01). The whole body muscle ratio showed negative correlation between lumbar lordosis (r=-0.28, p=0.02) and thoracic kyphosis angles (r=-0.33, p=0.00), although there was no statistically correlation between trunk muscle ratio, lumbar and thoracic curvatures (p>0.05). The study demonstrated that an increase of fat ratio and decrease of muscle ratio in abdominal region or whole body shifts the spinal alignment which may adversely affect the spinal loading. Therefore, whole body composition should be taken into account in spine rehabilitation.

Keywords: body composition, lumbar lordosis, spinal alignment, thoracic kyphosis

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3569 Comparison of Transparent Nickel Doped Cobalt Sulfide and Platinum Counter Electrodes Used in Quasi-Solid State Dye Sensitized Solar Cells

Authors: Dimitra Sygkridou, Dimitrios Karageorgopoulos, Elias Stathatos, Evangelos Vitoratos

Abstract:

Transparent nickel doped cobalt sulfide was fabricated on a SnO2:F electrode and tested as an efficient electrocatalyst and as an alternative to the expensive platinum counter electrode. In order to investigate how this electrode could affect the electrical characteristics of a dye-sensitized solar cell, we manufactured cells with the same TiO2 photoanode sensitized with dye (N719) and employing the same quasi-solid electrolyte, altering only the counter electrode used. The cells were electrically and electrochemically characterized and it was observed that the ones with the Ni doped CoS2 outperformed the efficiency of the cells with the Pt counter electrode (3.76% and 3.44% respectively). Particularly, the higher efficiency of the cells with the Ni doped CoS2 counter electrode (CE) is mainly because of the enhanced photocurrent density which is attributed to the enhanced electrocatalytic ability of the CE and the low charge transfer resistance at the CE/electrolyte interface.

Keywords: nickel doped cobalt sulfide, counter electrodes, dye-sensitized solar cells, quasi-solid state electrolyte, hybrid organic-inorganic materials

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3568 Performance and Lifetime of Tandem Organic Solar Cells

Authors: Guillaume Schuchardt, Solenn Berson, Gerard Perrier

Abstract:

Multi-junction solar cell configurations, where two sub-cells with complementary absorption are stacked and connected in series, offer an exciting approach to tackle the single junction limitations of organic solar cells and improve their power conversion efficiency. However, the augmentation of the number of layers has, as a consequence, to increase the risk of reducing the lifetime of the cell due to the ageing phenomena present at the interfaces. In this work, we study the intrinsic degradation mechanisms, under continuous illumination AM1.5G, inert atmosphere and room temperature, in single and tandem organic solar cells using Impedance Spectroscopy, IV Curves, External Quantum Efficiency, Steady-State Photocarrier Grating, Scanning Kelvin Probe and UV-Visible light.

Keywords: single and tandem organic solar cells, intrinsic degradation mechanisms, characterization: SKP, EQE, SSPG, UV-Visible, Impedance Spectroscopy, optical simulation

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3567 Accumulation of Phlorotannins in Abalone Haliotis discus Hannai after Feeding with Eisenia bicyclis

Authors: Bangoura Issa, Ji-Young Kang, M. T. H. Chowdhury, Ji-Eun Lee, Yong-Ki Hong

Abstract:

Investigation was carried out for the production of value-added abalone Haliotis discus hannai containing bioactive phlorotannin by feeding phlorotannin-rich seaweed Eisenia bicyclis 2 weeks prior to harvesting. Accumulation of phlorotannins was proceded by feeding with E. bicyclis after 4 days of starvation. HPLC purification afforded two major phlorotannins. Mass spectrometry and 1H-nuclear magnetic resonance analysis clarified their structures to be as 7-phloroeckol and eckol. Throughout the feeding period of 20 days, 7-phloroeckolol was accumulated in the muscle (foot muscle tissue) up to 0.18±0.12 mg g-1 dry weight of tissue after 12 days. Eckol reached 0.21±0.03 mg g-1 dry weight of tissue after 18 days. By feeding Laminaria japonica as reference, abalone showed no detection of phlorotannins in the muscle tissue. Seaweed consumption and growth rate of abalone revealed almost similar when feed with E. bicyclis or L. japonicain 20 days. Phlorotannins reduction to half-maximal accumulation values took 1.0 day and 2.7 days for 7-phloroeckol and eckol respectively, after replacing the feed to L. japonica.

Keywords: abalone, accumulation, eisenia bicyclis, phlorotannins

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3566 L-Carnitine Supplementation and Exercise-Induced Muscle Damage

Authors: B. Nakhostin-Roohi, F. Khoshkhahesh, KH. Parandak, R. Ramazanzadeh

Abstract:

Introduction: The protective effect of antioxidants in diminishing the post-exercise rise of serum CK and LDH in individuals trained for competitive sports has come to light in recent years. This study was conducted to assess the effect of Two-week L-carnitine supplementation on exercise-induced muscle damage, as well as antioxidant capacity after a bout of strenuous exercise in active healthy young men. Methodology: Twenty active healthy men volunteered for this study. Participants were randomized in a double-blind placebo-controlled fashion into two groups: L-carnitine (C group; n = 10) and placebo group (P group; n = 10). The participants took supplementation (2000 mg L-carnitine) or placebo (2000 mg lactose) daily for 2weeks before the main trial. Then, participants ran 14 km. Blood samples were taken before supplementation, before exercise, immediately, 2h and 24h after exercise. Creatine kinase (CK), and lactate dehydrogenase (LDH), and total antioxidant capacity (TAC) were measured. Results: Serum CK and LDH significantly increased after exercise in both groups (p < 0.05). Serum LDH was significantly lower in C group than P group 2h and 24h after exercise (p < 0.05). Furthermore, CK was significantly lower in C group compared with P group just 24h after exercise (p < 0.05). Plasma TAC increased significantly 14 days after supplementation and 24h after exercise in C group compared with P group (p < 0.05). Discussion and conclusion: These results suggest two-week daily oral supplementation of L-carnitine has been able to promote antioxidant capacity before and after exercise and decrease muscle damage markers through possibly inhibition of exercise-induced oxidative stress.

Keywords: L-carnitine, muscle damage, creatine kinase, Lactate dehydrogenase

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3565 Removal of Samarium in Environmental Water Samples by Modified Yeast Cells

Authors: Homayon Ahmad Panahi, Seyed Mehdi Seyed Nejad, Elham Moniri

Abstract:

A novel bio-adsorbent is fabricated by attaching a cibacron blue to yeast cells. The modified bio-sorbent has been characterized by some techniques like Fourier transform infrared spectroscopy (FT-IR) and elemental analysis (CHN) and applied for the preconcentration and determination of samarium from aqueous water samples. The best pH value for adsorption of the brilliant crecyle blue by yeast cells- cibacron blue was 7. The sorption capacity of modified biosorbent was 18.5 mg. g⁻¹. A recovery of 95.3% was obtained for Sm(III) when eluted with 0.5 M nitric acid. The method was applied for Sm(III) preconcentration and determination in river water sample.

Keywords: samarium, solid phase extraction, yeast cells, water sample, removal

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3564 A Case Report of Aberrant Vascular Anatomy of the Deep Inferior Epigastric Artery Flap

Authors: Karissa Graham, Andrew Campbell-Lloyd

Abstract:

The deep inferior epigastric artery perforator flap (DIEP) is used to reconstruct large volumes of tissue. The DIEP flap is based on the deep inferior epigastric artery (DIEA) and vein. Accurate knowledge of the anatomy of these vessels allows for efficient dissection of the flap, minimal damage to surrounding tissue, and a well vascularized flap. A 54 year old lady was assessed for bilateral delayed autologous reconstruction with DIEP free flaps. The right DIEA was consistent with the described anatomy. The left DIEA had a vessel branching shortly after leaving the external iliac artery and before entering the muscle. This independent branch entered the muscle and had a long intramuscular course to the largest perforator. The main DIEA vessel demonstrated a type II branching pattern but had perforators that were too small to have a viable DIEP flap. There were no communicating arterial branches between the independent vessel and DIEA, however, there was one venous communication between them. A muscle sparing transverse rectus abdominis muscle flap was raised using the main periumbilical perforator from the independent vessel. Our case report demonstrated an unreported anatomical variant of the DIEA. A few anatomical variants have been described in the literature, including a unilateral absent DIEA and peritoneal-cutaneous perforators that had no connection to the DIEA. Doing a pre-operative CTA helps to identify these rare anatomical variations, which leads to safer, more efficient, and effective operating.

Keywords: aberrant anatomy, CT angiography, DIEP anatomy, free flap

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3563 Assessment of Post-surgical Donor-Site Morbidity in Vastus lateralis Free Flap for Head and Neck Reconstructive Surgery: An Observational Study

Authors: Ishith Seth, Lyndel Hewitt, Takako Yabe, James Wykes, Jonathan Clark, Bruce Ashford

Abstract:

Background: Vastus lateralis (VL) can be used to reconstruct defects of the head and neck. Whilst the advantages are documented, donor-site morbidity is not well described. This study aimed to assess donor-site morbidity after VL flap harvest. The results will determine future directions for preventative and post-operative care to improve patient health outcomes. Methods: Ten participants (mean age 55 years) were assessed for the presence of donor-site morbidity after VL harvest. Musculoskeletal (pain, muscle strength, muscle length, tactile sensation), quality of life (SF-12), and lower limb function (lower extremity function, gait (function and speed), sit to stand were assessed using validated and standardized procedures. Outcomes were compared to age-matched healthy reference values or the non-operative side. Analyses were conducted using descriptive statistics and non-parametric tests. Results: There was no difference in muscle strength (knee extension), muscle length, ability to sit-to-stand, or gait function (all P > 0.05). Knee flexor muscle strength was significantly less on the operated leg compared to the non-operated leg (P=0.02) and walking speed was slower than age-matched healthy values (P<0.001). Thigh tactile sensation was impaired in 89% of participants. Quality of life was significantly less for the physical health component of the SF-12 (P<0.001). The mental health component of the SF-12 was similar to healthy controls (P=0.26). Conclusion: There was no effect on donor site morbidity with regards to knee extensor strength, pain, walking function, ability to sit-to-stand, and muscle length. VL harvest affected donor-site knee flexion strength, walking speed, tactile sensation, and physical health-related quality of life.

Keywords: vastus lateralis, morbidity, head and neck, surgery, donor-site morbidity

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3562 Viscoelastic Separation and Concentration of Candida Using a Low Aspect Ratio Microchannel

Authors: Seonggil Kim, Jeonghun Nam, Chae Seung Lim

Abstract:

Rapid diagnosis of fungal infections is critical for rapid antifungal therapy. However, it is difficult to detect extremely low concentration fungi in blood sample. To address the limitation, separation and concentration of fungi in blood sample are required to enhance the sensitivity of PCR analysis. In this study, we demonstrated a sheathless separation and concentration of fungi, candida cells using a viscoelastic fluid. To validate the performance of the device, microparticle mixture (2 and 13 μm) was used, and those particles were successfully separated based on the size difference at high flow rate of 100 μl/min. For the final application, successful separation of the Candida cells from the white blood cells (WBCs) was achieved. Based on the viscoelastic lateral migration toward the equilibrium position, Candida cells were separated and concentrated by center focusing, while WBCs were removed by patterning into two streams between the channel center and the sidewalls. By flow cytometric analysis, the separation efficiency and the purity were evaluated as ~99% and ~ 97%, respectively. From the results, the device can be the powerful tool for detecting extremely rare disease-related cells.

Keywords: candida cells, concentration, separation, viscoelastic fluid

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3561 Electromyography Pattern Classification with Laplacian Eigenmaps in Human Running

Authors: Elnaz Lashgari, Emel Demircan

Abstract:

Electromyography (EMG) is one of the most important interfaces between humans and robots for rehabilitation. Decoding this signal helps to recognize muscle activation and converts it into smooth motion for the robots. Detecting each muscle’s pattern during walking and running is vital for improving the quality of a patient’s life. In this study, EMG data from 10 muscles in 10 subjects at 4 different speeds were analyzed. EMG signals are nonlinear with high dimensionality. To deal with this challenge, we extracted some features in time-frequency domain and used manifold learning and Laplacian Eigenmaps algorithm to find the intrinsic features that represent data in low-dimensional space. We then used the Bayesian classifier to identify various patterns of EMG signals for different muscles across a range of running speeds. The best result for vastus medialis muscle corresponds to 97.87±0.69 for sensitivity and 88.37±0.79 for specificity with 97.07±0.29 accuracy using Bayesian classifier. The results of this study provide important insight into human movement and its application for robotics research.

Keywords: electromyography, manifold learning, ISOMAP, Laplacian Eigenmaps, locally linear embedding

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3560 Increased Cytolytic Activity of Effector T-Cells against Cholangiocarcinoma Cells by Self-Differentiated Dendritic Cells with Down-Regulation of Interleukin-10 and Transforming Growth Factor-β Receptors

Authors: Chutamas Thepmalee, Aussara Panya, Mutita Junking, Jatuporn Sujjitjoon, Nunghathai Sawasdee, Pa-Thai Yenchitsomanus

Abstract:

Cholangiocarcinoma (CCA) is an aggressive malignancy of bile duct epithelial cells in which the standard treatments, including surgery, radiotherapy, chemotherapy, and targeted therapy are partially effective. Many solid tumors including CCA escape host immune responses by creating tumor microenvironment and generating immunosuppressive cytokines such as interleukin-10 (IL-10) and transforming growth factor-β (TGF-β). These cytokines can inhibit dendritic cell (DC) differentiation and function, leading to decreased activation and response of effector CD4+ and CD8+ T cells for cancer cell elimination. To overcome the effects of these immunosuppressive cytokines and to increase ability of DC to activate effector CD4+ and CD8+ T cells, we generated self-differentiated DCs (SD-DCs) with down-regulation of IL-10 and TGF-β receptors for activation of effector CD4+ and CD8+ T cells. Human peripheral blood monocytes were initially transduced with lentiviral particles containing the genes encoding GM-CSF and IL-4 and then secondly transduced with lentiviral particles containing short-hairpin RNAs (shRNAs) to knock-down mRNAs of IL-10 and TGF-β receptors. The generated SD-DCs showed up-regulation of MHC class II (HLA-DR) and co-stimulatory molecules (CD40 and CD86), comparable to those of DCs generated by convention method. Suppression of IL-10 and TGF-β receptors on SD-DCs by specific shRNAs significantly increased levels of IFN-γ and also increased cytolytic activity of DC-activated effector T cells against CCA cell lines (KKU-213 and KKU-100), but it had little effect to immortalized cholangiocytes (MMNK-1). Thus, SD-DCs with down-regulation of IL-10 and TGF-β receptors increased activation of effector T cells, which is a recommended method to improve DC function for the preparation of DC-activated effector T cells for adoptive T-cell therapy.

Keywords: cholangiocarcinoma, IL-10 receptor, self-differentiated dendritic cells, TGF-β receptor

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3559 Based on MR Spectroscopy, Metabolite Ratio Analysis of MRI Images for Metastatic Lesion

Authors: Hossain A, Hossain S.

Abstract:

Introduction: In a small cohort, we sought to assess the magnetic resonance spectroscopy's (MRS) ability to predict the presence of metastatic lesions. Method: A Popular Diagnostic Centre Limited enrolled patients with neuroepithelial tumors. The 1H CSI MRS of the brain allows us to detect changes in the concentration of specific metabolites caused by metastatic lesions. Among these metabolites are N-acetyl-aspartate (NNA), creatine (Cr), and choline (Cho). For Cho, NAA, Cr, and Cr₂, the metabolic ratio was calculated using the division method. Results: The NAA values were 0.63 and 5.65 for tumor cells, 1.86 and 5.66 for normal cells, and 1.86 and 5.66 for normal cells 2. NAA values for normal cells 1 were 1.84, 10.6, and 1.86 for normal cells 2, respectively. Cho levels were as low as 0.8 and 10.53 in the tumor cell, compared to 1.12 and 2.7 in the normal cell 1 and 1.24 and 6.36 in the normal cell 2. Cho/Cr₂ barely distinguished itself from the other ratios in terms of significance. For tumor cells, the ratios of Cho/NAA, Cho/Cr₂, NAA/Cho, and NAA/Cr₂ were significant. Normal cell 1 had significant Cho/NAA, Cho/Cr, NAA/Cho, and NAA/Cr ratios. Conclusion: The clinical result can be improved by using 1H-MRSI to guide the size of resection for metastatic lesions. Even though it is non-invasive and doesn't present any difficulties during the procedure, MRS has been shown to predict the detection of metastatic lesions.

Keywords: metabolite ratio, MRI images, metastatic lesion, MR spectroscopy, N-acetyl-aspartate

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3558 Evaluation of Gene Expression after in Vitro Differentiation of Human Bone Marrow-Derived Stem Cells to Insulin-Producing Cells

Authors: Mahmoud M. Zakaria, Omnia F. Elmoursi, Mahmoud M. Gabr, Camelia A. AbdelMalak, Mohamed A. Ghoneim

Abstract:

Many protocols were publicized for differentiation of human mesenchymal stem cells (MSCS) into insulin-producing cells (IPCs) in order to excrete insulin hormone ingoing to treat diabetes disease. Our aim is to evaluate relative gene expression for each independent protocol. Human bone marrow cells were derived from three volunteers that suffer diabetes disease. After expansion of mesenchymal stem cells, differentiation of these cells was done by three different protocols (the one-step protocol was used conophylline protein, the two steps protocol was depending on trichostatin-A, and the three-step protocol was started by beta-mercaptoethanol). Evaluation of gene expression was carried out by real-time PCR: Pancreatic endocrine genes, transcription factors, glucose transporter, precursor markers, pancreatic enzymes, proteolytic cleavage, extracellular matrix and cell surface protein. Quantitation of insulin secretion was detected by immunofluorescence technique in 24-well plate. Most of the genes studied were up-regulated in the in vitro differentiated cells, and also insulin production was observed in the three independent protocols. There were some slight increases in expression of endocrine mRNA of two-step protocol and its insulin production. So, the two-step protocol was showed a more efficient in expressing of pancreatic endocrine genes and its insulin production than the other two protocols.

Keywords: mesenchymal stem cells, insulin producing cells, conophylline protein, trichostatin-A, beta-mercaptoethanol, gene expression, immunofluorescence technique

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3557 Chemical Speciation and Bioavailability of Some Essential Metal Ions In Different Fish Organs at Lake Chamo, Ethiopia

Authors: Adane Gebresilassie Hailemariam, Belete Yilma Hirpaye

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The enhanced concentrations of heavy metals, especially in sediments, may indicate human-induced perturbations rather than natural enrichment through geological weathering. Heavy metals are non-biodegradable, persist in the environment, and are concentrated up to the food chain, leading to enhanced levels in the liver and muscle tissues of fishes, aquatic bryophytes, and aquatic biota. Marine organisms, in general fish in particular, accumulate metals to concentrations many times higher than present in water or sediment as they can take up metals in their organs and concentrate at different levels. Thus, metals acquired through the food chain due to pollution are potential chemical hazards, threatening consumers. The Nile tilapia (oreochromic niloticus), catfish (clarius garpinus), and water samples were collected from five sampling sites, namely, inlet-1, inlet-2, center, outlet-1 and outlet-2 of Lake Chamo. The concentration of major and trace metals Na, K, Mg, Ca, Cr, Co, Ni, Mn and Cu in the two fish muscles, gill and liver, was determined using an atomic absorption spectrometer (AAS) and flame photometer (FP). Metal concentrations in the water have also been evaluated within the two consecutive seasons, winter (dry) and spring (wet). The results revealed that the concentration of those metals in Tilapia’s (O. niloticus) muscle, gill, and liver were Na 44.5, 35.1, 28, Mg 2.8, 8.41, 4.61, K 43, 32, 30, Ca 1.5, 6.0, 5.5, Cr 0.91, 1.2, 3.5, Co 3.0, 2.89, 2.62, Ni 0.94, 1.99, 2.2, Mn 1.23, 1.51, 1.6 and Cu 1.1, 1.99, 3.5 mg kg-1 respectively and in catfish’s muscle, gill and liver Na 25, 39, 41.5, Mg 4.8, 2.87, 6, K 29, 38, 40, Ca 2.5, 8.10, 3.0, Cr 0.65, 3.5, 5.0, Co 2.62, 1.86, 1.73, Ni 1.10, 2.3, 3.1, Mn 1.54, 1.57, 1.59 and Cu 1.01, 1.10, 3.70 mg kg-1 respectively. The highest accumulation of Na and K were observed for tilapia muscle and catfish gill, Mg and Ca got higher in tilapia gill and catfish liver, while Co is higher in muscle of the two fish. The Cr, Ni, Mn and Cu levels were higher in the livers of the two fish species. In conculusion, metal toxicity through food chain is the current dangerous issue for human and othe animals. This needs deep focus to promot the health of living animals. The Details of the work are going to be discussed at the conference.

Keywords: bioaccumulation, catfish, essential metals, nile tilapia

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3556 Current Concepts of Male Aesthetics: Facial Areas to Be Focused and Prioritized with Botulinum Toxin and Hyaluronic Acid Dermal Fillers Combination Therapies, Recommendations on Asian Patients

Authors: Sadhana Deshmukh

Abstract:

Objective: Men represent only a fraction of the medical aesthetic practice. They are increasingly becoming more cosmetically-inclined. The primary objective is to harmonize facial proportion by prioritizing and focusing on forehead nose, cheek and chin complex. Introduction: Despite tremendous variability, diverse population of the Indian subcontinent, the male skull is unique in its overall larger size, and shape. Men tend to have a large forehead with prominent supraorbital ridges, wide glabella, square orbit, and a prominent protruding mandible. Men have increased skeletal muscle mass, with less facial subcutaneous fat. Facial aesthetics is evolving rapidly. Commonly published canons of facial proportions usually represent feminine standards and are not applicable to males. Strict adherence to these norms is therefore not necessary to obtain satisfying results in male patients. Materials and Methods: Male patients age group 30-60 years have been enrolled. Botulinum toxin and hyaluronic acid fillers were used to update consensus recommendations for facial rejuvenation using these two types of products alone and in combination. Results: There are specific recommendations by facial area, focusing on relaxing musculature, restoring volume, recontouring using toxin and dermal fillers alone and in combination. For upper face, though botulinum toxin remains the cornerstone of treatment, temples and forehead fillers are recommended for optimal results. In Mid face, these fillers are placed more laterally to maintain the masculine look. Botulinum toxin and fillers in combination can improve outcomes in the lower face. Chin augmentation remains the center point for lower face. Conclusions: Males are more likely to have shorter doctor visits, less likely to ask questions, have a lower attention to bodily changes. The physician must patiently gauge male patients’ aging and cosmetic goals. Clinicians can also benefit from ongoing guidance on products, tailoring treatments, treating multiple facial areas, and using combinations of products. An appreciation that rejuvenation is 3-dimensional process involving muscle control, volume restoration and recontouring helps.

Keywords: male aesthetics, botulinum toxin, hyaluronic acid dermal fillers, Asian patients

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3555 A Novel Application of CORDYCEPIN (Cordycepssinensis Extract): Maintaining Stem Cell Pluripotency and Improving iPS Generation Efficiency

Authors: Shih-Ping Liu, Cheng-Hsuan Chang, Yu-Chuen Huang, Shih-Yin Chen, Woei-Cherng Shyu

Abstract:

Embryonic stem cells (ES) and induced pluripotnet stem cells (iPS) are both pluripotent stem cells. For mouse stem cells culture technology, leukemia inhibitory factor (LIF) was used to maintain the pluripotency of stem cells in vitro. However, LIF is an expensive reagent. The goal of this study was to find out a pure compound extracted from Chinese herbal medicine that could maintain stem cells pluripotency to replace LIF and improve the iPS generation efficiency. From 20 candidates traditional Chinese medicine we found that Cordycepsmilitaris triggered the up-regulation of stem cells activating genes (Oct4 and Sox2) expression levels in MEF cells. Cordycepin, a major active component of Cordycepsmilitaris, also could up-regulate Oct4 and Sox2 gene expression. Furthermore, we used ES and iPS cells and treated them with different concentrations of Cordycepin (replaced LIF in the culture medium) to test whether it was useful to maintain the pluripotency. The results showed higher expression levels of several stem cells markers in 10 μM Cordycepin-treated ES and iPS cells compared to controls that did not contain LIF, including alkaline phosphatase, SSEA1, and Nanog. Embryonic body formation and differentiation confirmed that 10 μM Cordycepin-containing medium was capable to maintain stem cells pluripotency after four times passages. For mechanism analysis, microarray analysis indicated extracellular matrix and Jak/Stat signaling pathway as the top two deregulated pathways. In ECM pathway, we determined that the integrin αVβ5 expression levels and phosphorylated Src levels increased after Cordycepin treatment. In addition, the phosphorylated Jak2 and phosphorylated Sat3 protein levels were increased after Cordycepin treatment and suppressed with the Jak2 inhibitor, AG490. The expression of cytokines associated with Jak2/Stat3 signaling pathway were also up-regulated by Q-PCR and ELISA assay. Lastly, we used Oct4-GFP MEF cells to test iPS generation efficiency following Cordycepin treatment. We observed that 10 Μm Cordycepin significantly increased the iPS generation efficiency in day 21. In conclusion, we demonstrated Cordycepin could maintain the pluripotency of stem cells through both of ECM and Jak2/Stat3 signaling pathway and improved iPS generation efficiency.

Keywords: cordycepin, iPS cells, Jak2/Stat3 signaling pathway, molecular biology

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3554 The Comparison between bFGF and Small Molecules in Derivation of Chicken Primordial Germ Cells and Embryonic Germ Cells

Authors: Maryam Farzaneh, Seyyedeh Nafiseh Hassani, Hossein Baharvand

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Objective: Chicken gonadal tissue has a two population such primordial germ cells (PGCs) and stromal cells (somatic cells). PGCs and embryonic germ cells (EGCs) that is a pluripotent type of PGCs in long-term culture are suitable sources for the production of chicken pluripotent stem cell lines, transgenic birds, vaccine and recombinant protein production. In general, the effect of growth factors such bFGF and mouse LIF on derivation of PGCs in vitro are important and in this study we could see the unique effect of small molecules such PD032 and SB43 as a chemical, in comparison to growth factors. Materials and Methods: After incubation of fertilized chicken egg up to 6 days and isolation of primary gonadal tissues and culture of mixed cells like PGCs and stromal cells. PGCs proliferate in the present of fetal calf serum (FCS) and small molecules and in another group bFGF, that these factors are important for PGCs culture and derivation. Somatic cells produce a multilayer feeder under the PGCs in primary culture and PGCs make a small cluster under these cells. Results: In present of small molecules and high volume of FCS (15%), the present of EGCs as a pluripotent stem cells were clear four weeks, that they had a positive immune-staining and periodic acid-Schiff staining (PAS), but in present of growth factors like bFGF without any chemicals, the present of PGCs were clear but after 7 until 10 days, there were disappear. Conclusion: Until now we have seen many researches about derivation and maintenance of chicken PGCs, in the hope of understanding the mechanisms that occur during germline development and production of a therapeutic product by transgenic birds. There are still many unknowns in this area and this project will try to have efficient conditions for identification of suitable culture medium for long-term culture of PGCs in vitro without serum and feeder cells.

Keywords: chicken gonadal primordial germ cells, pluripotent stem cells, growth factors, small molecules, transgenic birds

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3553 An Ergonomic Evaluation of Three Load Carriage Systems for Reducing Muscle Activity of Trunk and Lower Extremities during Giant Puppet Performing Tasks

Authors: Cathy SW. Chow, Kristina Shin, Faming Wang, B. C. L. So

Abstract:

During some dynamic giant puppet performances, an ergonomically designed load carrier system is necessary for the puppeteers to carry a giant puppet body’s heavy load with minimum muscle stress. A load carrier (i.e. prototype) was designed with two small wheels on the foot; and a hybrid spring device on the knee in order to assist the sliding and knee bending movements respectively. Thus, the purpose of this study was to evaluate the effect of three load carriers including two other commercially available load mounting systems, Tepex and SuitX, and the prototype. Ten male participants were recruited for the experiment. Surface electromyography (sEMG) was used to collect the participants’ muscle activities during forward moving and bouncing and with and without load of 11.1 kg that was 60 cm above the shoulder. Five bilateral muscles including the lumbar erector spinae (LES), rectus femoris (RF), bicep femoris (BF), tibialis anterior (TA), and gastrocnemius (GM) were selected for data collection. During forward moving task, the sEMG data showed smallest muscle activities by Tepex harness which exhibited consistently the lowest, compared with the prototype and SuitX which were significantly higher on left LES 68.99% and 64.99%, right LES 26.57% and 82.45%; left RF 87.71% and 47.61%, right RF 143.57% and 24.28%; left BF 80.21% and 22.23%, right BF 96.02% and 21.83%; right TA 6.32% and 4.47%; left GM 5.89% and 12.35% respectively. The result above reflected mobility was highly restricted by tested exoskeleton devices. On the other hand, the sEMG data from bouncing task showed the smallest muscle activities by prototype which exhibited consistently the lowest, compared with the Tepex harness and SuitX which were significantly lower on lLES 6.65% and 104.93, rLES 23.56% and 92.19%; lBF 33.21% and 93.26% and rBF 24.70% and 81.16%; lTA 46.51% and 191.02%; rTA 12.75% and 125.76%; IGM 31.54% and 68.36%; rGM 95.95% and 96.43% respectively.

Keywords: exoskeleton, giant puppet performers, load carriage system, surface electromyography

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3552 Chemical Modification of Biosorbent for Prconcentation of Cadmium in Water Sample

Authors: Homayon Ahmad Panahi, Niusha Mohseni Darabi, Elham Moniri

Abstract:

A new biosorbent is prepared by coupling a cibacron blue to yeast cells. The modified yeast cells with cibacron blue has been characterized by Fourier transform infrared spectroscopy (FT-IR) and elemental analysis and applied for the preconcentration and solid phase extraction of trace cadmium ion from water samples. The optimum pH value for sorption of the cadmium ions by yeast cells- cibacron blue was 5.5. The sorption capacity of modified biosorbent was 45 mg. g−1. A recovery of 98.2% was obtained for Cd(II) when eluted with 0.5 M nitric acid. The method was applied for Cd(II) preconcentration and determination in sea water sample.

Keywords: solid phase extraction, yeast cells, Nickl, isotherm study

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3551 The Role of Il-6-Mediated NS5ATP9 Expression in Autophagy of Liver Cancer Cells

Authors: Hongping Lu, Kelbinur Tursun, Yaru Li, Yu Zhang, Shunai Liu, Ming Han

Abstract:

Objective: To investigate whether NS5ATP9 is involved in IL-6 mediated autophagy and the relationship between IL-6 and NS5ATP9 in liver cancer cells. Methods: 1. Detect the mRNA and protein levels of Beclin 1 after HepG2 cells were treated with or without recombinant human IL-6 protein. 2. Measure and compare of the changes of autophagy-related genes with their respective control, after IL-6 was silenced or neutralized with monoclonal antibody against human IL-6. 3. HepG2 cells were incubated with 50 ng/ml of IL-6 in the presence or absence of PDTC. The expression of NS5ATP9 was analyzed by Western blot after 48 h. 4. After NS5ATP9-silenced HepG2 cells had been treated with 50 ng/ml recombinant IL-6 protein, we detected the Beclin 1 and LC3B (LC3Ⅱ/Ⅰ) expression. 5. HepG2 cells were transfected with pNS5ATP9, si-NS5ATP9, and their respective control. Total RNA was isolated from cells and analyzed for IL-6. 6. Silence or neutralization of IL-6 in HepG2 cells which has been transfected with NS5ATP9. Beclin 1 and LC3 protein levels were analyzed by Western blot. Result: 1. After HepG2 were treated with recombinant human IL-6 protein, the expression of endogenous Beclin 1 was up-regulated at mRNA and protein level, and the conversion of endogenous LC3-I to LC3-II was also increased. These results indicated that IL-6 could induce autophagy. 2. When HepG2 cells were treated with IL-6 siRNA or monoclonal antibody against human IL-6, the expression of autophagy-related genes were decreased. 3. Exogenous human IL-6 recombinant protein up-regulated NS5ATP9 via NF-κB activation. 4. The expression of Beclin 1 and LC3B was down-regulated after IL-6 treated NS5ATP9-silenced HepG2 cells. 5. NS5ATP9 could reverse regulates IL-6 expression in HepG2 cells. 6. Silence or neutralization of IL-6 attenuates NS5ATP9-induced autophagy slightly. Conclusion: Our results implied that in HCC patients, maybe the higher level of IL-6 in the serum promoted the expression of NS5ATP9 and induced autophagy in cancer cells. And the over-expression of NS5ATP9 which induced by IL-6, in turn, increased IL-6 expression, further, promotes the IL-6/NS5ATP9-mediated autophagy and affects the progression of tumor. Therefore, NS5ATP9 silence might be a potential target for HCC therapy.

Keywords: autophagy, Hepatocellular carcinoma, IL-6, microenvironment, NS5ATP9

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3550 Aerobic Exercise Increases Circulating Hematopoietic Stem Cells and Endothelial Progenitor Cells

Authors: Khaled A. shady, Fagr B. Bazeed, Nashwa K. Abousamra, Ihab H. Elberawe, Ashraf E. shaalan, Mohamed A. Sobh

Abstract:

Physical activity activates a variety of adult stem cells which might be released into the circulation or might be activated in their organ-resident state. A variety of stimuli such as metabolic, mechanical, and hormonal stimuli might by responsible for the mobilization. This study was done to know the changes in hematopoietic stem cells and endothelial progenitor in athletes in the 24 hours following 30 min of aerobic exercise. Methods: Ten healthy male's athlete's (age 20.7± 0.61 y) performed moderate running with 30 min at 80% of velocity of The IAT. Blood samples taken pre-, and immediately, 30 min, 2h, 6h and 24h post-exercise were analyzed for hematopoietic stem cells (HSCs ), endothelial progenitor cells (EPCs(, vascular endothelial growth factor (VEGF), nitric oxide (NO), lactic acid (LA), and white blood cells . HSCs and EPCs were quantified by flow cytometry. Results: After 30min of aerobic exercise significant increases in HSCs, EPC, VEGF, NO, LA and WBCs (p ˂ 0.05). This increase will be at different rates according to the timing of taking blood sample and was in the maximum rate of increase after 30 min of aerobic exercise. HSCs, EPC, NO and WBCs were in the maximum rate of increase 2h post exercise. In addition, VEGF was in the maximum rate of increase immediately post exercise and LA concentration not affected after exercise. Conclusion: These data suggest that HSCs and EPCs increased after aerobic exercise due to increase of VEGF which play an important role in mobilization of stem cells and promotes NO increase which contributes to increase EPCs.

Keywords: physical activity, hematopoietic stem cells, mobilization, athletes

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3549 Teratogenic Effect of Bisphenol A in Development of Balb/C Mouse

Authors: Nazihe Sedighi, Mohsen Nokhbatolphoghaei

Abstract:

Bisphenol A (BPA) is a monomer used in the manufacture of polycarbonate plastics. Due to having properties such as transparency, heat and impact resistance, it is used widely in medicine, sorts, electronic components, and food containers. It is also used in the production of resins which is applied for lining cans. BPA releases from resins and polycarbonate when it is heated or continuously used the containers from which BPA can enter the body. There are several reports indicating the presence of BPA in the placenta, amniotic fluid, and the embryo itself. While researchers investigated the teratogenic effect of BPA on embryos, very limited work has been done on the effects of BPA when applied from early stages of development. In this study, The teratogenic effect of BPA was investigated at earliest preimplantation (day zero) through day 15.5 of the development of Balb/C mouse embryos. After ensuring the pregnancy via observing vaginal plug, Pregnant mice were divided into five groups. For the three experimental groups, the amount of 500, 750, and 1000 mg/kg/d Bisphenol A was given orally according to body weight. The sham group that was treated with sesame oil, which was used as vehicle and control group remained intact. On day 18.5 of gestation, embryos were removed from the uterus. Randomly half of the embryo were fixed in Bouin for tissue analysis. The other half were prepared for skeletal system staining using Alizarin Red and alcian blue dies. The results showed that the embryonic weight and the crown-rump length of embryos decreased significantly (P < 0.05) in all experimental groups compared to the control group and the sham. In this study, skeletal abnormalities such as delay in ossification of skull and limbs as well as the deviation in the backbone were seen. This research suggests that pregnant mothers need to be aware of possible teratogenic effects of BPA at any stage of pregnancy especially from early to mid stages. In this case, pregnant mothers may need to stop using any manufacture of polycarbonate plastics, as a container for food or drinking.

Keywords: bisphenol A, development, polycarbonate plastic, skeletal system, teratogenicity

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3548 The Effect of Mesenchymal Stem Cells on Full Thickness Skin Wound Healing in Albino Rats

Authors: Abir O. El Sadik

Abstract:

Introduction: Wound healing involves the interaction of multiple biological processes among different types of cells, intercellular matrix and specific signaling factors producing enhancement of cell proliferation of the epidermis over dermal granulation tissue. Several studies investigated multiple strategies to promote wound healing and to minimize infection and fluid losses. However, burn crisis, and its related morbidity and mortality are still elevated. The aim of the present study was to examine the effects of mesenchymal stem cells (MSCs) in accelerating wound healing and to compare the most efficient route of administration of MSCs, either intradermal or systemic injection, with focusing on the mechanisms producing epidermal and dermal cell regeneration. Material and methods: Forty-two adult male Sprague Dawley albino rats were divided into three equal groups (fourteen rats in each group): control group (group I); full thickness surgical skin wound model, Group II: Wound treated with systemic injection of MSCs and Group III: Wound treated with intradermal injection of MSCs. The healing ulcer was examined on day 2, 6, 10 and 15 for gross morphological evaluation and on day 10 and 15 for fluorescent, histological and immunohistochemical studies. Results: The wounds of the control group did not reach complete closure up to the end of the experiment. In MSCs treated groups, better and faster healing of wounds were detected more than the control group. Moreover, the intradermal route of administration of stem cells increased the rate of healing of the wounds more than the systemic injection. In addition, the wounds were found completely healed by the end of the fifteenth day of the experiment in all rats of the group injected intradermally. Microscopically, the wound areas of group III were hardly distinguished from the adjacent normal skin with complete regeneration of all skin layers; epidermis, dermis, hypodermis and underlying muscle layer. Fully regenerated hair follicles and sebaceous glands in the dermis of the healed areas surrounded by different arrangement of collagen fibers with a significant increase in their area percent were recorded in this group more than in other groups. Conclusion: MSCs accelerate the healing process of wound closure. The route of administration of MSCs has a great influence on wound healing as intradermal injection of MSCs was more effective in enhancement of wound healing than systemic injection.

Keywords: intradermal, mesenchymal stem cells, morphology, skin wound, systemic injection

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3547 Graphene Materials for Efficient Hybrid Solar Cells: A Spectroscopic Investigation

Authors: Mohammed Khenfouch, Fokotsa V. Molefe, Bakang M. Mothudi

Abstract:

Nowadays, graphene and its composites are universally known as promising materials. They show their potential in a large field of applications including photovoltaics. This study reports on the role of nanohybrids and nanosystems known as strong light harvesters in the efficiency of graphene hybrid solar cells. Our system included Graphene/ZnO/Porphyrin/P3HT layers. Moreover, the physical properties including surface/interface, optical and vibrational properties were also studied. Our investigations confirmed the interaction between the different components as well as the sensitivity of their photonics to the synthesis conditions. Remarkable energy and charge transfer were detected and deeply investigated. Hence, the optimization of the conditions will lead to the fabrication of higher conversion efficiency in graphene solar cells.

Keywords: graphene, optoelectronics, nanohybrids, solar cells

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3546 Induction of Apoptosis by Diosmin through Interleukins/STAT and Mitochondria Mediated Pathway in Hep-2 and KB Cells

Authors: M. Rajasekar, K. Suresh

Abstract:

Diosmin is a flavonoid, most abundantly found in many citrus fruits. As a flavonoid, it possesses a multitude of biological activities including anti-hyperglycemic, anti-lipid peroxidative, anti-inflammatory, antioxidant, and anti-mutagenic properties. At this point, we established the anti-proliferative and apoptosis-inducing activities of diosmin in Hep-2 and KB cells. Diosmin has cytotoxic effects through inhibiting cellular proliferation of Hep-2 and KB cells, which leads to the induction of apoptosis, as apparent by an increase in the fraction of cells in the sub-G1phase of the cell cycle. Results exposed that inhibition of cell proliferation is associated with regulation of the Interleukins/STAT pathway. In addition, Diosmin treatment with Hep-2 and KB cells actively stimulated reactive oxygen species (ROS) and mitochondrial membrane depolarization. And also an imbalance in the Bax/Bcl-2 ratio triggered the caspase cascade and shifting the balance in favor of apoptosis. These observations conclude that Diosmin induce apoptosis via Interleukins /STAT-mediated pathway.

Keywords: diosmin, apoptosis, antioxidant, STAT pathway

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3545 Following the Modulation of Transcriptional Activity of Genes by Chromatin Modifications during the Cell Cycle in Living Cells

Authors: Sharon Yunger, Liat Altman, Yuval Garini, Yaron Shav-Tal

Abstract:

Understanding the dynamics of transcription in living cells has improved since the development of quantitative fluorescence-based imaging techniques. We established a method for following transcription from a single copy gene in living cells. A gene tagged with MS2 repeats, used for mRNA tagging, in its 3' UTR was integrated into a single genomic locus. The actively transcribing gene was detected and analyzed by fluorescence in situ hybridization (FISH) and live-cell imaging. Several cell clones were created that differed in the promoter regulating the gene. Thus, comparative analysis could be obtained without the risk of different position effects at each integration site. Cells in S/G2 phases could be detected exhibiting two adjacent transcription sites on sister chromatids. A sharp reduction in the transcription levels was observed as cells progressed along the cell cycle. We hypothesized that a change in chromatin structure acts as a general mechanism during the cell cycle leading to down-regulation in the activity of some genes. We addressed this question by treating the cells with chromatin decondensing agents. Quantifying and imaging the treated cells suggests that chromatin structure plays a role both in regulating transcriptional levels along the cell cycle, as well as in limiting an active gene from reaching its maximum transcription potential at any given time. These results contribute to understanding the role of chromatin as a regulator of gene expression.

Keywords: cell cycle, living cells, nucleus, transcription

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3544 Induction of G1 Arrest and Apoptosis in Human Cancer Cells by Panaxydol

Authors: Dong-Gyu Leem, Ji-Sun Shin, Sang Yoon Choi, Kyung-Tae Lee

Abstract:

In this study, we focused on the anti-proliferative effects of panaxydol, a C17 polyacetylenic compound derived from Panax ginseng roots, against various human cancer cells. We treated with panaxydol to various cancer cells and panaxydol treatment was found to significantly inhibit the proliferation of human lung cancer cells (A549) and human pancreatic cancer cells (AsPC-1 and MIA PaCa-2), of which AsPC-1 cells were most sensitive to its treatment. DNA flow cytometric analysis indicated that panaxydol blocked cell cycle progression at the G1 phase in A549 cells, which accompanied by a parallel reduction of protein expression of cyclin-dependent kinase (CDK) 2, CDK4, CDK6, cyclin D1 and cyclin E. CDK inhibitors (CDKIs), such as p21CIP1/WAF1 and p27KIP1, were gradually upregulated after panaxydol treatment at the protein levels. Furthermore, panaxydol induced the activation of p53 in A549 cells. In addition, panaxydol also induced apoptosis of AsPC-1 and MIA PaCa-2 cells, as shown by accumulation of subG1 and apoptotic cell populations. Panaxydol triggered the activation of caspase-3, -8, -9 and the cleavage of poly (ADP-ribose) polymerase (PARP). Reduction of mitochondrial transmembrane potential by panaxydol was determined by staining with dihexyloxacarbocyanine iodide. Furthermore, panaxydol suppressed the levels of anti-apoptotic proteins, XIAP and Bcl-2, and increased the levels of proapoptotic proteins, Bax and Bad. In addition, panaxydol inhibited the activation of Akt and extracellular signal-regulated kinase (ERK) and activated the p38 mitogen-activated protein kinase kinase (MAPK). Our results suggest that panaxydol is an anti-tumor compound that causes p53-mediated cell cycle arrest and apoptosis via mitochondrial apoptotic pathway in various cancer cells.

Keywords: apoptosis, cancer, G1 arrest, panaxydol

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3543 Lipid-polymer Nanocarrier Platform Enables X-Ray Induced Photodynamic Therapy against Human Colorectal Cancer Cells

Authors: Rui Sang, Fei Deng, Alexander Engel, Ewa M. Goldys, Wei Deng

Abstract:

In this study, we brought together X-ray induced photodynamic therapy (X-PDT) and chemo-drug (5-FU) for the treatment on colorectal cancer cells. This was achieved by developing a lipid-polymer hybrid nanoparticle delivery system (FA-LPNPs-VP-5-FU). It was prepared by incorporating a photosensitizer (verteporfin), chemotherapy drug (5-FU), and a targeting moiety (folic acid) into one platform. The average size of these nanoparticles was around 100 nm with low polydispersity. When exposed to clinical doses of 4 Gy X-ray radiation, FA-LPNPs-VP-5-FU generated sufficient amounts of reactive oxygen species, triggering the apoptosis and necrosis pathway of cancer cells. Our combined X-PDT and chemo-drug strategy was effective in inhibiting cancer cells’ growth and proliferation. Cell cycle analyses revealed that our treatment induced G2/M and S phase arrest in HCT116 cells. Our results indicate that this combined treatment provides better antitumour effect in colorectal cancer cells than each of these modalities alone. This may offer a novel approach for effective colorectal cancer treatment with reduced off-target effect and drug toxicity.

Keywords: pdt, targeted lipid-polymer nanoparticles, verteporfin, colorectal cancer

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3542 Effect of Nitrogen-Based Cryotherapy on the Calf Muscle Spasticity in Stroke Patients

Authors: Engi E. I. Sarhan, Usama M. Rashad, Ibrahim M. I. Hamoda, Mohammed K. Mohamed

Abstract:

Background: This study aimed to know the effect of nitrogen-based cryotherapy on the spasticity of calf muscle in stroke patients. Patients were selected from the outpatient clinic of Neurology, Al-Mansoura general hospital, Al-Mansoura University. Subjects and methods: Thirty Stroke Patients of both sexes ranged from 45 to 60 years old were divided randomly into two equal groups, a study group (A) received a nitrogen-based cryotherapy, a selective physical therapy program and ankle foot orthosis (AFO), while as patients in control group (B) received the same program and AFO only. The treatment duration was three times per week for four weeks for both groups. We assessed spasticity of calf muscle before and after treatment subjectively using modified Ashworth scale (MAS) and objectively via measuring H / M ratio on electromyography machine. We also assessed ankle dorsiflexion ROM objectively using two dimensions motion analysis (2D). Results: After treatment, there was a highly significant improvement in the study group compared to the control group regarding the score of MAS, no significant difference in the study group compared to the control group regarding the readings of H / M ratio, highly significant improvement in the study group compared to the control group regarding the 2D motion analysis findings. Conclusion: This modality considers effective in reducing spasticity in the calf muscle and improving ankle dorsiflexion of the affected limb.

Keywords: ankle foot orthosis, nitrogen-based cryotherapy, stroke, spasticity

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3541 Hyaluronic Acid - Alginate Hydrogel for the Transdifferentiation of Testis Cells into Erythrocyte and Hepatocyte-like Cells; A Practice Within an Effective Agent Choice

Authors: Leila Rashki Ghaleno, Mohamad Amin Hajari, Leila Montazeri, Abdolhossein Shahverdi, Mojtaba Rezazadeh Valojerdi

Abstract:

Background: Spermatogonia stem cells (SSCs) exhibit pluripotency, enabling them to undergo differentiation into many cell lineages, including neurons, glia, endothelial cells, and hepatocytes when cultured in vitro. Although the specific mechanisms are not yet fully understood, it has been observed that biopolymer agents, such as hyaluronic acid (HA) and alginate (Alg), have the potential to induce transdifferentiation of SSCs. The current work aimed to examine the process of in vitro spermatogenesis and the conversion of mouse testicular cells into hepatocytes and erythrocyte-like cells utilizing the HA-Alg hydrogel. Method: After being extracted from the testes of a 5-day postpartum mouse (5 DPP), the testicular cells were separated into two enzymatic stages and then put into a composite hydrogel containing 0.5% HA and 1% alginate. On days 14 and 28 of culture, the colonies' growth, the cells' viability, and their histology were assessed. Result: Despite observing significant cell proliferation on day 14 and the development of circular-shaped organoids on day 28, it was noted that the organoids generated in the HA-Alg medium tended to maintain their circular morphology on day 28. Notably, the testicular cells underwent transdifferentiation into cell types resembling erythrocytes and hepatocytes. The hepatocyte-like cells exhibited the presence of glycogen and lipid deposits, indicating their hepatocyte-like characteristics. Interestingly, immunostaining analysis revealed the secretion of albumin and the presence of VEGFR on day 14. However, on day 28, albumin expression was not detected, while the expression of Sox9 (a marker for hepatocytes), Vegf, CD34, and C-kit (markers for erythrocytes) showed increased levels in the gene expression evaluation. Conclusion: The present findings indicated that HA-Alg could be a potent and effective agent for the transdifferentiation of testis cells into erythrocyte and hepatocyte-like cells, as recent studies have confirmed the transformation of SSCs into hepatocyte cells during in vitro culture.

Keywords: 3D culture, mouse testicular cell, hyaluronic acid, liver organoids

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