Search results for: muscle stem cells
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 4366

Search results for: muscle stem cells

3796 Measurement System for Human Arm Muscle Magnetic Field and Grip Strength

Authors: Shuai Yuan, Minxia Shi, Xu Zhang, Jianzhi Yang, Kangqi Tian, Yuzheng Ma

Abstract:

The precise measurement of muscle activities is essential for understanding the function of various body movements. This work aims to develop a muscle magnetic field signal detection system based on mathematical analysis. Medical research has underscored that early detection of muscle atrophy, coupled with lifestyle adjustments such as dietary control and increased exercise, can significantly enhance muscle-related diseases. Currently, surface electromyography (sEMG) is widely employed in research as an early predictor of muscle atrophy. Nonetheless, the primary limitation of using sEMG to forecast muscle strength is its inability to directly measure the signals generated by muscles. Challenges arise from potential skin-electrode contact issues due to perspiration, leading to inaccurate signals or even signal loss. Additionally, resistance and phase are significantly impacted by adipose layers. The recent emergence of optically pumped magnetometers introduces a fresh avenue for bio-magnetic field measurement techniques. These magnetometers possess high sensitivity and obviate the need for a cryogenic environment unlike superconducting quantum interference devices (SQUIDs). They detect muscle magnetic field signals in the range of tens to thousands of femtoteslas (fT). The utilization of magnetometers for capturing muscle magnetic field signals remains unaffected by issues of perspiration and adipose layers. Since their introduction, optically pumped atomic magnetometers have found extensive application in exploring the magnetic fields of organs such as cardiac and brain magnetism. The optimal operation of these magnetometers necessitates an environment with an ultra-weak magnetic field. To achieve such an environment, researchers usually utilize a combination of active magnetic compensation technology with passive magnetic shielding technology. Passive magnetic shielding technology uses a magnetic shielding device built with high permeability materials to attenuate the external magnetic field to a few nT. Compared with more layers, the coils that can generate a reverse magnetic field to precisely compensate for the residual magnetic fields are cheaper and more flexible. To attain even lower magnetic fields, compensation coils designed by Biot-Savart law are involved to generate a counteractive magnetic field to eliminate residual magnetic fields. By solving the magnetic field expression of discrete points in the target region, the parameters that determine the current density distribution on the plane can be obtained through the conventional target field method. The current density is obtained from the partial derivative of the stream function, which can be represented by the combination of trigonometric functions. Optimization algorithms in mathematics are introduced into coil design to obtain the optimal current density distribution. A one-dimensional linear regression analysis was performed on the collected data, obtaining a coefficient of determination R2 of 0.9349 with a p-value of 0. This statistical result indicates a stable relationship between the peak-to-peak value (PPV) of the muscle magnetic field signal and the magnitude of grip strength. This system is expected to be a widely used tool for healthcare professionals to gain deeper insights into the muscle health of their patients.

Keywords: muscle magnetic signal, magnetic shielding, compensation coils, trigonometric functions.

Procedia PDF Downloads 57
3795 A Multi Function Myocontroller for Upper Limb Prostheses

Authors: Ayad Asaad Ibrahim

Abstract:

Myoelectrically controlled prostheses are becoming more and more popular, for below-elbow amputation, the wrist flexor and extensor muscle group, while for above-elbow biceps and triceps brachii muscles are used for control of the prosthesis. A two site multi-function controller is presented. Two stainless steel bipolar electrode pairs are used to monitor the activities in both muscles. The detected signals are processed by new pre-whitening technique to identify the accurate tension estimation in these muscles. These estimates will activate the relevant prosthesis control signal, with a time constant of 200 msec. It is ensured that the tension states in the control muscle to activate a particular prosthesis function are similar to those used to activate normal functions in the natural hand. This facilitates easier training.

Keywords: prosthesis, biosignal processing, pre-whitening, myoelectric controller

Procedia PDF Downloads 363
3794 Technique and Use of Machine Readable Dictionary: In Special Reference to Hindi-Marathi Machine Translation

Authors: Milind Patil

Abstract:

Present paper is a discussion on Hindi-Marathi Morphological Analysis and generating rules for Machine Translation on the basis of Machine Readable Dictionary (MRD). This used Transformative Generative Grammar (TGG) rules to design the MRD. As per TGG rules, the suffix of a particular root word is based on its Tense, Aspect, Modality and Voice. That's why the suffix is very important for the word meanings (or root meanings). The Hindi and Marathi Language both have relation with Indo-Aryan language family. Both have been derived from Sanskrit language and their script is 'Devnagari'. But there are lots of differences in terms of semantics and grammatical level too. In Marathi, there are three genders, but in Hindi only two (Masculine and Feminine), the Natural gender is absent in Hindi. Likewise other grammatical categories also differ in their level of use. For MRD the suffixes (or Morpheme) are of particular root word for GNP (Gender, Number and Person) are based on its natural phenomena. A particular Suffix and Morphine change as per the need of person, number and gender. The design of MRD also based on this format. In first, Person, Number, Gender and Tense are key points than root words and suffix of particular Person, Number Gender (PNG). After that the inferences are drawn on the basis of rules that is (V.stem) (Pre.T/Past.T) (x) + (Aux-Pre.T) (x) → (V.Stem.) + (SP.TM) (X).

Keywords: MRD, TGG, stem, morph, morpheme, suffix, PNG, TAM&V, root

Procedia PDF Downloads 325
3793 Improved Wearable Monitoring and Treatment System for Parkinson’s Disease

Authors: Bulcha Belay Etana, Benny Malengier, Janarthanan Krishnamoorthy, Timothy Kwa, Lieva VanLangenhove

Abstract:

Electromyography measures the electrical activity of muscles using surface electrodes or needle electrodes to monitor various disease conditions. Recent developments in the signal acquisition of electromyograms using textile electrodes facilitate wearable devices, enabling patients to monitor and control their health status outside of healthcare facilities. Here, we have developed and tested wearable textile electrodes to acquire electromyography signals from patients suffering from Parkinson’s disease and incorporated a feedback-control system to relieve muscle cramping through thermal stimulus. In brief, the textile electrodes made of stainless steel was knitted into a textile fabric as a sleeve, and their electrical characteristic, such as signal-to-noise ratio, was compared with traditional electrodes. To relieve muscle cramping, a heating element made of stainless-steel conductive yarn sewn onto cotton fabric, coupled with a vibration system, was developed. The system integrated a microcontroller and a Myoware muscle sensor to activate the heating element as well as the vibration motor when cramping occurs, and at the same time, the element gets deactivated when the muscle cramping subsides. An optimum therapeutic temperature of 35.5 °C is regulated by continuous temperature monitoring to deactivate the heating system when this threshold value is reached. The textile electrode exhibited a signal-to-noise ratio of 6.38dB, comparable to that of the traditional electrode’s value of 7.05 dB. For a given 9 V power supply, the rise time was about 6 minutes for the developed heating element to reach an optimum temperature.

Keywords: smart textile system, wearable electronic textile, electromyography, heating textile, vibration therapy, Parkinson’s disease

Procedia PDF Downloads 108
3792 Enhancing Power Conversion Efficiency of P3HT/PCBM Polymer Solar Cells

Authors: Nidal H. Abu-Zahra, Mahmoud Algazzar

Abstract:

In this research, n-dodecylthiol was added to P3HT/PC70BM polymer solar cells to improve the crystallinity of P3HT and enhance the phase separation of P3HT/PC70BM. The improved crystallinity of P3HT/PC70BM doped with 0-5% by volume of n-dodecylthiol resulted in improving the power conversion efficiency of polymer solar cells by 33%. In addition, thermal annealing of the P3HT/PC70MB/n-dodecylthiolcompound showed further improvement in crystallinity with n-dodecylthiol concentration up to 2%. The highest power conversion efficiency of 3.21% was achieved with polymer crystallites size L of 11.2nm, after annealing at 150°C for 30 minutes under a vacuum atmosphere. The smaller crystallite size suggests a shorter path of the charge carriers between P3HT backbones, which could be beneficial to getting a higher short circuit current in the devices made with the additive.

Keywords: n-dodecylthiol, congugated PSC, P3HT/PCBM, polymer solar cells

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3791 Annona muricata Leaves Induced Mitochondrial-Mediated Apoptosis in A549 Cells

Authors: Soheil Zorofchian Moghadamtousi, Habsah Abdul Kadir, Mohammadjavad Paydar, Elham Rouhollahi, Hamed Karimian

Abstract:

The present study was designed to evaluate the molecular mechanisms of Annona muricata leaves ethyl acetate extract (AMEAE) against lung cancer A549 cells. Cell viability analysis revealed the selective cytotoxic effect of AMEAE towards A549 cells. Treatment of A549 cells with AMEAE significantly elevated the reactive oxygen species formation, followed by attenuation of mitochondrial membrane potential via upregulation of Bax and downregulation of Bcl-2, accompanied by cytochrome c release to the cytosol. The released cytochrome c triggered the activation of caspase-9 followed by caspase-3. In addition, AMEAE-induced apoptosis was accompanied by cell cycle arrest at G1 phase. Our data showed for the first time that AMEAE inhibited the proliferation of A549 cells, leading to cell cycle arrest and programmed cell death through activation of the mitochondrial-mediated signaling pathway.

Keywords: Annona muricata, lung cancer, apoptosis, mitochondria

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3790 Viscoelastic Cell Concentration in a High Aspect Ratio Microchannel Using a Non-Powered Air Compressor

Authors: Jeonghun Nam, Seonggil Kim, Hyunjoo Choi, Chae Seung Lim

Abstract:

Quantification and analysis of rare cells are challenging in clinical applications and cell biology due to its extremely small number in blood. In this work, we propose a viscoelastic microfluidic device for continuous cell concentration without sheath flows. Due to the viscoelastic effect on suspending cells, cells with the blockage ratio higher than 0.1 could be tightly focused at the center of the microchannel. The blockage ratio was defined as the particle diameter divided by the channel width. Finally, cells were concentrated through the center outlet and the additional suspending medium was removed to the side outlets. Since viscoelastic focusing is insensitive to the flow rate higher than 10 μl/min, the non-powered hand pump sprayer could be used with no accurate control of the flow rate, which is suitable for clinical settings in resource-limited developing countries. Using multiple concentration processes, high-throughput concentration of white blood cells in lysed blood sample was achieved by ~ 300-fold.

Keywords: cell concentration, high-throughput, non-powered, viscoelastic fluid

Procedia PDF Downloads 288
3789 Greywater Reuse for Sunflower Irrigation Previously Radiated with Helium-Neon Laser: Evaluation of Growth, Flowering, and Chemical Constituents

Authors: Sami Ali Metwally, Bedour Helmy Abou-Leila, Hussien Ibrahim Abdel-Shafy

Abstract:

This study was carried out at the pilot plant area in the National Research Centre during the two successive seasons, 2020 and 2022. The aim is to investigate the response of vegetative growth and chemical constituents of sunflowers plants irrigated by two types of wastewater, namely: black wastewater W1 (Bathroom) and grey wastewater W1, under irradiation conditions of helium-neon (He-Ne) laser. The examined data indicated that irrigation of W1 significantly increased the growth and flowering parameters (plant height, leaves number, leaves area, leaves fresh and dry weight, flower diameter, flower stem length, flower stem thickness, number of days to flower, and total chlorophyll). Treated sunflower plants with 0 to 10 min. recorded an increase in the fresh weight and dry weight of leaves. However, the superiority of increasing vase life and delaying flowers were recorded by prolonging exposure time by up to 10 min. Regarding the effect of interaction treatments, the data indicated that the highest values on almost growth parameters were obtained from plants treated with W1+0 laser followed by W2+10 min. laser, compared with all interaction treatments. As for flowering parameters, the interactions between W2+2 min. time exposure, W1+0 time, w1+10 min., and w1+2 min. exposures recorded the highest values on flower diameter, flower stem length, flower stem thickness, vase life, and delaying flowering.

Keywords: greywater, sunflower plant, water reuse, vegetative growth, laser radiation

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3788 Human Brain Organoids-on-a-Chip Systems to Model Neuroinflammation

Authors: Feng Guo

Abstract:

Human brain organoids, 3D brain tissue cultures derived from human pluripotent stem cells, hold promising potential in modeling neuroinflammation for a variety of neurological diseases. However, challenges remain in generating standardized human brain organoids that can recapitulate key physiological features of a human brain. Here, this study presents a series of organoids-on-a-chip systems to generate better human brain organoids and model neuroinflammation. By employing 3D printing and microfluidic 3D cell culture technologies, the study’s systems enable the reliable, scalable, and reproducible generation of human brain organoids. Compared with conventional protocols, this study’s method increased neural progenitor proliferation and reduced heterogeneity of human brain organoids. As a proof-of-concept application, the study applied this method to model substance use disorders.

Keywords: human brain organoids, microfluidics, organ-on-a-chip, neuroinflammation

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3787 Determination of Circulating Tumor Cells in Breast Cancer Patients by Electrochemical Biosensor

Authors: Gökçe Erdemir, İlhan Yaylım, Serap Erdem-Kuruca, Musa Mutlu Can

Abstract:

It has been determined that the main reason for the death of cancer disease is caused by metastases rather than the primary tumor. The cells that leave the primary tumor and enter the circulation and cause metastasis in the secondary organs are called "circulating tumor cells" (CTCs). The presence and number of circulating tumor cells has been associated with poor prognosis in many major types of cancer, including breast, prostate, and colorectal cancer. It is thought that knowledge of circulating tumor cells, which are seen as the main cause of cancer-related deaths due to metastasis, plays a key role in the diagnosis and treatment of cancer. The fact that tissue biopsies used in cancer diagnosis and follow-up are an invasive method and are insufficient in understanding the risk of metastasis and the progression of the disease have led to new searches. Liquid biopsy tests performed with a small amount of blood sample taken from the patient for the detection of CTCs are easy and reliable, as well as allowing more than one sample to be taken over time to follow the prognosis. However, since these cells are found in very small amounts in the blood, it is very difficult to capture them and specially designed analytical techniques and devices are required. Methods based on the biological and physical properties of the cells are used to capture these cells in the blood. Early diagnosis is very important in following the prognosis of tumors of epithelial origin such as breast, lung, colon and prostate. Molecules such as EpCAM, vimentin, and cytokeratins are expressed on the surface of cells that pass into the circulation from very few primary tumors and reach secondary organs from the circulation, and are used in the diagnosis of cancer in the early stage. For example, increased EpCAM expression in breast and prostate cancer has been associated with prognosis. These molecules can be determined in some blood or body fluids to be taken from patients. However, more sensitive methods are required to be able to determine when they are at a low level according to the course of the disease. The aim is to detect these molecules found in very few cancer cells with the help of sensitive, fast-sensing biosensors, first in breast cancer cells reproduced in vitro and then in blood samples taken from breast cancer patients. In this way, cancer cells can be diagnosed early and easily and effectively treated.

Keywords: electrochemical biosensors, breast cancer, circulating tumor cells, EpCAM, Vimentin, Cytokeratins

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3786 Mirna Expression Profile is Different in Human Amniotic Mesenchymal Stem Cells Isolated from Obese Respect to Normal Weight Women

Authors: Carmela Nardelli, Laura Iaffaldano, Valentina Capobianco, Antonietta Tafuto, Maddalena Ferrigno, Angela Capone, Giuseppe Maria Maruotti, Maddalena Raia, Rosa Di Noto, Luigi Del Vecchio, Pasquale Martinelli, Lucio Pastore, Lucia Sacchetti

Abstract:

Maternal obesity and nutrient excess in utero increase the risk of future metabolic diseases in the adult life. The mechanisms underlying this process are probably based on genetic, epigenetic alterations and changes in foetal nutrient supply. In mammals, the placenta is the main interface between foetus and mother, it regulates intrauterine development, modulates adaptive responses to sub optimal in uterus conditions and it is also an important source of human amniotic mesenchymal stem cells (hA-MSCs). We previously highlighted a specific microRNA (miRNA) profiling in amnion from obese (Ob) pregnant women, here we compared the miRNA expression profile of hA-MSCs isolated from (Ob) and control (Co) women, aimed to search for any alterations in metabolic pathways that could predispose the new-born to the obese phenotype. Methods: We isolated, at delivery, hA-MSCs from amnion of 16 Ob- and 7 Co-women with pre-pregnancy body mass index (mean/SEM) 40.3/1.8 and 22.4/1.0 kg/m2, respectively. hA-MSCs were phenotyped by flow cytometry. Globally, 384 miRNAs were evaluated by the TaqMan Array Human MicroRNA Panel v 1.0 (Applied Biosystems). By the TargetScan program we selected the target genes of the miRNAs differently expressed in Ob- vs Co-hA-MSCs; further, by KEGG database, we selected the statistical significant biological pathways. Results: The immunophenotype characterization confirmed the mesenchymal origin of the isolated hA-MSCs. A large percentage of the tested miRNAs, about 61.4% (232/378), was expressed in hA-MSCs, whereas 38.6% (146/378) was not. Most of the expressed miRNAs (89.2%, 207/232) did not differ between Ob- and Co-hA-MSCs and were not further investigated. Conversely, 4.8% of miRNAs (11/232) was higher and 6.0% (14/232) was lower in Ob- vs Co-hA-MSCs. Interestingly, 7/232 miRNAs were obesity-specific, being expressed only in hA-MSCs isolated from obese women. Bioinformatics showed that these miRNAs significantly regulated (P<0.001) genes belonging to several metabolic pathways, i.e. MAPK signalling, actin cytoskeleton, focal adhesion, axon guidance, insulin signaling, etc. Conclusions: Our preliminary data highlight an altered miRNA profile in Ob- vs Co-hA-MSCs and suggest that an epigenetic miRNA-based mechanism of gene regulation could affect pathways involved in placental growth and function, thereby potentially increasing the newborn’s risk of metabolic diseases in the adult life.

Keywords: hA-MSCs, obesity, miRNA, biosystem

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3785 Impact of Helicobacter pylori Infection on Colorectal Adenoma-Colorectal Carcinoma Sequence

Authors: Jannis Kountouras, Nikolaos Kapetanakis, Stergios A. Polyzos, Apostolis Papaeftymiou, Panagiotis Katsinelos, Ioannis Venizelos, Christina Nikolaidou, Christos Zavos, Iordanis Romiopoulos, Elena Tsiaousi, Evangelos Kazakos, Michael Doulberis

Abstract:

Background & Aims: Helicobacter pylori infection (Hp-I) has been recognized as a substantial risk agent involved in gastrointestinal (GI) tract oncogenesis by stimulating cancer stem cells (CSCs), oncogenes, immune surveillance processes, and triggering GI microbiota dysbiosis. We aimed to investigate the possible involvement of active Hp-I in the sequence: chronic inflammation–adenoma–colorectal cancer (CRC) development. Methods: Four pillars were investigated: (i) endoscopic and conventional histological examinations of patients with CRC, colorectal adenomas (CRA) versus controls to detect the presence of active Hp-I; (ii) immunohistochemical determination of the presence of Hp; expression of CD44, an indicator of CSCs and/or bone marrow-derived stem cells (BMDSCs); expressions of oncogene Ki67 and anti-apoptotic Bcl-2 protein; (iii) expression of CD45, indicator of immune surveillance locally (assessing mainly T and B lymphocytes locally); and (iv) correlation of the studied parameters with the presence or absence of Hp-I. Results: Among 50 patients with CRC, 25 with CRA, and 10 controls, a significantly higher presence of Hp-I in the CRA (68%) and CRC group (84%) were found compared with controls (30%). The presence of Hp-I with accompanying immunohistochemical expression of CD44 in biopsy specimens was revealed in a high proportion of patients with CRA associated with moderate/severe dysplasia (88%) and CRC patients with moderate/severe degree of malignancy (91%). Comparable results were also obtained for Ki67, Bcl-2, and CD45 immunohistochemical expressions. Concluding Remarks: Hp-I seems to be involved in the sequence: CRA – dysplasia – CRC, similarly to the upper GI tract oncogenesis, by several pathways such as the following: Beyond Hp-I associated insulin resistance, the major underlying mechanism responsible for the metabolic syndrome (MetS) that increase the risk of colorectal neoplasms, as implied by other Hp-I related MetS pathologies, such as non-alcoholic fatty liver disease and upper GI cancer, the disturbance of the normal GI microbiota (i.e., dysbiosis) and the formation of an irritative biofilm could contribute to a perpetual inflammatory upper GIT and colon mucosal damage, stimulating CSCs or recruiting BMDSCs and affecting oncogenes and immune surveillance processes. Further large-scale relative studies with a pathophysiological perspective are necessary to demonstrate in-depth this relationship.

Keywords: Helicobacter pylori, colorectal cancer, colorectal adenomas, gastrointestinal oncogenesis

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3784 The Influence of Alginate Microspheres Modified with DAT on the Proliferation and Adipogenic Differentiation of ASCs

Authors: Shin-Yi Mao, Jiashing Yu

Abstract:

Decellularized adipose tissue (DAT) has received lots of attention as biological scaffolds recently. DAT that extracted from the extracellular matrix (ECM) of adipose tissues holds great promise as a xenogeneic biomaterial for tissue engineering and regenerative medicine. In our study, 2-D DATsol film was fabricated to enhance cell adhesion, proliferation, and differentiation of ASCs in vitro. DAT was also used to modify alginate for improvement of cell adhesion. Alginate microspheres modified with DAT were prepared by Nisco. These microspheres could provide a highly supportive 3-D environment for ASCs. In our works, ASCs were immobilized in alginate microspheres modified with DAT to promoted cell adhesion and adipogenic differentiation. Accordingly, we hypothesize that tissue regeneration in vivo could be promoted with the aid of modified microspheres in future.

Keywords: adipose stem cells, decellularize adipose tissue, Alginate, microcarries

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3783 Synergistic Cytotoxicity of Cisplatin and Taxol in Overcoming Taxol Resistance through the Inhibition of LDHA in Oral Squamous Cell Carcinoma

Authors: Lin Feng, Ling-Ling E., Hong-Chen Liu

Abstract:

The development of chemoresistance in patients represents a major challenge in cancer treatment. Lactate dehydrogenase‑A (LDHA) is one of the principle isoforms of LDH that is expressed in breast tissue, controlling the conversion of pyruvate to lactate and also playing a significant role in the metabolism of glucose. The aim of this study was to identify whether LDHA was involved in oral cancer cell resistance to Taxol and whether the downregulation of LDHA, as a result of cisplatin treatment, may overcome Taxol resistance in human oral squamous cells. The OECM‑1 oral epidermal carcinoma cell line was used, which has been widely used as a model of oral cancer in previous studies. The role of LDHA in Taxol and cisplatin resistance was investigated and the synergistic cytotoxicity of cisplatin and/or Taxol in oral squamous cells was analyzed. Cell viability was analyzed by MTT assay, LDHA expression was analyzed by western blot analysis and siRNA transfection was performed to knock down LDHA expression. The present study results showed that decreased levels of LDHA were responsible for the resistance of oral cancer cells to cisplatin (CDDP). CDDP treatments downregulated LDHA expression and lower levels of LDHA were detected in the CDDP‑resistant oral cancer cells compared with the CDDP‑sensitive cells. By contrast, the Taxol‑resistant cancer cells showed elevated LDHA expression levels. In addition, small interfering RNA‑knockdown of LDHA sensitized the cells to Taxol but desensitized them to CDDP treatment while exogenous expression of LDHA sensitized the cells to CDDP, but desensitized them to Taxol. The present study also revealed the synergistic cytotoxicity of CDDP and Taxol for killing oral cancer cells through the inhibition of LDHA. This study highlights LDHA as a novel therapeutic target for overcoming Taxol resistance in oral cancer patients using the combined treatments of Taxol and CDDP.

Keywords: cisplatin, Taxol, carcinoma, oral squamous cells

Procedia PDF Downloads 419
3782 Evaluation of Mito-Uncoupler Induced Hyper Metabolic and Aggressive Phenotype in Glioma Cells

Authors: Yogesh Rai, Saurabh Singh, Sanjay Pandey, Dhananjay K. Sah, B. G. Roy, B. S. Dwarakanath, Anant N. Bhatt

Abstract:

One of the most common signatures of highly malignant gliomas is their capacity to metabolize more glucose to lactic acid than normal brain tissues, even under normoxic conditions (Warburg effect), indicating that aerobic glycolysis is constitutively upregulated through stable genetic or epigenetic changes. However, oxidative phosphorylation (OxPhos) is also required to maintain the mitochondrial membrane potential for tumor cell survival. In the process of tumorigenesis, tumor cells during fastest growth rate exhibit both high glycolytic and high OxPhos. Therefore, metabolically reprogrammed cancer cells with combination of both aerobic glycolysis and altered OxPhos develop a robust metabolic phenotype, which confers a selective growth advantage. In our study, we grew the high glycolytic BMG-1 (glioma) cells with continuous exposure of mitochondrial uncoupler 2, 4, dinitro phenol (DNP) for 10 passages to obtain a phenotype of high glycolysis with enhanced altered OxPhos. We found that OxPhos modified BMG (OPMBMG) cells has similar growth rate and cell cycle distribution but high mitochondrial mass and functional enzymatic activity than parental cells. In in-vitro studies, OPMBMG cells showed enhanced invasion, proliferation and migration properties. Moreover, it also showed enhanced angiogenesis in matrigel plug assay. Xenografted tumors from OPMBMG cells showed reduced latent period, faster growth rate and nearly five folds reduction in the tumor take in nude mice compared to BMG-1 cells, suggesting that robust metabolic phenotype facilitates tumor formation and growth. OPMBMG cells which were found radio-resistant, showed enhanced radio-sensitization by 2-DG as compared to the parental BMG-1 cells. This study suggests that metabolic reprogramming in cancer cells enhances the potential of migration, invasion and proliferation. It also strengthens the cancer cells to escape the death processes, conferring resistance to therapeutic modalities. Our data also suggest that combining metabolic inhibitors like 2-DG with conventional therapeutic modalities can sensitize such metabolically aggressive cancer cells more than the therapies alone.

Keywords: 2-DG, BMG, DNP, OPM-BMG

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3781 Branched Chain Amino Acid Kinesio PVP Gel Tape from Extract of Pea (Pisum sativum L.) Based on Ultrasound-Assisted Extraction Technology

Authors: Doni Dermawan

Abstract:

Modern sports competition as a consequence of the increase in the value of the business and entertainment in the field of sport has been demanding athletes to always have excellent physical endurance performance. Physical exercise is done in a long time, and intensive may pose a risk of muscle tissue damage caused by the increase of the enzyme creatine kinase. Branched Chain Amino Acids (BCAA) is an essential amino acid that is composed of leucine, isoleucine, and valine which serves to maintain muscle tissue, keeping the immune system, and prevent further loss of coordination and muscle pain. Pea (Pisum sativum L.) is a kind of leguminous plants that are rich in Branched Chain Amino Acids (BCAA) where every one gram of protein pea contains 82.7 mg of leucine; 56.3 mg isoleucine; and 56.0 mg of valine. This research aims to develop Branched Chain Amino Acids (BCAA) from pea extract is applied in dosage forms Gel PVP Kinesio Tape technology using Ultrasound-assisted Extraction. The method used in the writing of this paper is the Cochrane Collaboration Review that includes literature studies, testing the quality of the study, the characteristics of the data collection, analysis, interpretation of results, and clinical trials as well as recommendations for further research. Extraction of BCAA in pea done using ultrasound-assisted extraction technology with optimization variables includes the type of solvent extraction (NaOH 0.1%), temperature (20-250C), time (15-30 minutes) power (80 watt) and ultrasonic frequency (35 KHz). The advantages of this extraction method are the level of penetration of the solvent into the membrane of the cell is high and can increase the transfer period so that the BCAA substance separation process more efficient. BCAA extraction results are then applied to the polymer PVP (Polyvinylpyrrolidone) Gel powder composed of PVP K30 and K100 HPMC dissolved in 10 mL of water-methanol (1: 1) v / v. Preparations Kinesio Tape Gel PVP is the BCAA in the gel are absorbed into the muscle tissue, and joints through tensile force then provides stimulation to the muscle circulation with variable pressure so that the muscle can increase the biomechanical movement and prevent damage to the muscle enzyme creatine kinase. Analysis and evaluation of test preparation include interaction, thickness, weight uniformity, humidity, water vapor permeability, the levels of the active substance, content uniformity, percentage elongation, stability testing, release profile, permeation in vitro and in vivo skin irritation testing.

Keywords: branched chain amino acid, BCAA, Kinesio tape, pea, PVP gel, ultrasound-assisted extraction

Procedia PDF Downloads 289
3780 The Effect of Resistance and Progressive Training on Hsp 70 and Glucose

Authors: F. Nameni, H. Poursadra

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The present study investigated resistance and progressive training alters the expression of chaperone proteins. These proteins function to maintain homeostasis, facilitate repair from injury, and provide protection. Nineteen training female in 2 groups taking part in the intervention volunteered to give blood samples. Levels of chaperone proteins were measured in response to resistance and progressive training. Hsp 70 levels were increased immediately after 2 h progressive training but decreased after resistance training. The data showed that human skeletal muscle responds to the stress of a single period of progressive training by up-regulating and resistance training by down-regulating expression of HSP70. Physical exercise can elevate core temperature and muscle temperatures and the expression pattern of HSP70 due to training status may be attributed to adaptive mechanisms.

Keywords: resistance training, heat shock proteins, leukocytes, Hsp 70

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3779 Effect of Ambient Oxygen Content and Lifting Frequency on the Participant’s Lifting Capabilities, Muscle Activities, and Perceived Exertion

Authors: Atef M. Ghaleb, Mohamed Z. Ramadan, Khalid Saad Aljaloud

Abstract:

The aim of this study is to assesses the lifting capabilities of persons experiencing hypoxia. It also examines the behavior of the physiological response induced through the lifting process related to changing in the hypoxia and lifting frequency variables. For this purpose, the study performed two consecutive tests by using; (1) training and acclimatization; and (2) an actual collection of data. A total of 10 male students from King Saud University, Kingdom of Saudi Arabia, were recruited in the study. A two-way repeated measures design, with two independent variables (ambient oxygen (15%, 18% and 21%)) and lifting frequency (1 lift/min and 4 lifts/min) and four dependent variables i.e., maximum acceptable weight of lift (MAWL), Electromyography (EMG) of four muscle groups (anterior deltoid, trapezius, biceps brachii, and erector spinae), rating of perceived exertion (RPE), and rating of oxygen feeling (ROF) were used in this study. The results show that lifting frequency has significantly impacted the MAWL and muscles’ activities. The oxygen content had a significant effect on the RPE and ROE. The study has revealed that acclimatization and training sessions significantly reduce the effect of the hypoxia on the human physiological parameters during the manual materials handling tasks.

Keywords: lifting capabilities, muscle activities, oxygen content, perceived exertion

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3778 Effect of Plasma Radiation on Keratinocyte Cells Involved in the Wound Healing Process

Authors: B. Fazekas, I. Korolov, K. Kutasi

Abstract:

Plasma medicine, which involves the use of gas discharge plasmas for medical applications is a rapidly growing research field. The use of non-thermal atmospheric pressure plasmas in dermatology to assist tissue regeneration by improving the healing of infected and/or chronic wounds is a promising application. It is believed that plasma can activate cells, which are involved in the wound closure. Non-thermal atmospheric plasmas are rich in chemically active species (such as O and N-atoms, O2(a) molecules) and radiative species such as the NO, N2+ and N2 excited molecules, which dominantly radiate in the 200-500 nm spectral range. In order to understand the effect of plasma species, both of chemically active and radiative species on wound healing process, the interaction of physical plasma with the human skin cells is necessary. In order to clarify the effect of plasma radiation on the wound healing process we treated keratinocyte cells – that are one of the main cell types in human skin epidermis – covered with a layer of phosphate-buffered saline (PBS) with a low power atmospheric pressure plasma. For the generation of such plasma we have applied a plasma needle. Here, the plasma is ignited at the tip of the needle in flowing helium gas in contact with the ambient air. To study the effect of plasma radiation we used a plasma needle configuration, where the plasma species – chemically active radicals and charged species – could not reach the treated cells, but only the radiation. For the comparison purposes, we also irradiated the cells using a UV-B light source (FS20 lamp) with a 20 and 40 mJ cm-2 dose of 312 nm. After treatment the viability and the proliferation of the cells have been examined. The proliferation of cells has been studied with a real time monitoring system called Xcelligence. The results have indicated, that the 20 mJ cm-2 dose did not affect cell viability, whereas the 40 mJ cm-2 dose resulted a decrease in cell viability. The results have shown that the plasma radiation have no quantifiable effect on the cell proliferation as compared to the non-treated cells.

Keywords: UV radiation, non-equilibrium gas discharges (non-thermal plasmas), plasma emission, keratinocyte cells

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3777 Micromechanical Compatibility Between Cells and Scaffold Mediates the Efficacy of Regenerative Medicine

Authors: Li Yang, Yang Song, Martin Y. M. Chiang

Abstract:

Objective: To experimentally substantiate the micromechanical compatibility between cell and scaffold, in the regenerative medicine approach for restoring bone volume, is essential for phenotypic transitions Methods: Through nanotechnology and electrospinning process, nanofibrous scaffolds were fabricated to host dental follicle stem cells (DFSCs). Blends (50:50) of polycaprolactone (PCL) and silk fibroin (SF), mixed with various content of cellulose nanocrystals (CNC, up to 5% in weight), were electrospun to prepare nanofibrous scaffolds with heterogeneous microstructure in terms of fiber size. Colloidal probe atomic force microscopy (AFM) and conventional uniaxial tensile tests measured the scaffold stiffness at the micro-and macro-scale, respectively. The cell elastic modulus and cell-scaffold adhesive interaction (i.e., a chemical function) were examined through single-cell force spectroscopy using AFM. The quantitative reverse transcription-polymerase chain reaction (qRT-PCR) was used to determine if the mechanotransduction signal (i.e., Yap1, Wwr2, Rac1, MAPK8, Ptk2 and Wnt5a) is upregulated by the scaffold stiffness at the micro-scale (cellular scale). Results: The presence of CNC produces fibrous scaffolds with a bimodal distribution of fiber diameter. This structural heterogeneity, which is CNC-composition dependent, remarkably modulates the mechanical functionality of scaffolds at microscale and macroscale simultaneously, but not the chemical functionality (i.e., only a single material property is varied). In in vitro tests, the osteogenic differentiation and gene expression associated with mechano-sensitive cell markers correlate to the degree of micromechanical compatibility between DFSCs and the scaffold. Conclusion: Cells require compliant scaffolds to encourage energetically favorable interactions for mechanotransduction, which are converted into changes in cellular biochemistry to direct the phenotypic evolution. The micromechanical compatibility is indeed important to the efficacy of regenerative medicine.

Keywords: phenotype transition, scaffold stiffness, electrospinning, cellulose nanocrystals, single-cell force spectroscopy

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3776 Poly(ε-Caprolactone)-Based Bilayered Scaffolds Prepared by Electrospinning for Tissue Engineering of Small-Diameter Vascular Grafts

Authors: Mohammed Fayez Al Rez

Abstract:

Nowadays, there is an unmet clinical need for new small-diameter vascular grafts to overcome the drawbacks of traditional methods used for treatment of widespread cardiovascular diseases. Vascular tissue engineering (VTE) is a promising approach that can be utilized to develop viable vascular grafts by in vitro seeding of functional cells onto a scaffold allowing them to attach, proliferate and differentiate. To achieve this purpose, the scaffold should provide cells with the initial necessary extracellular matrix environment and structure until being able to reconstruct the required vascular tissue. Therefore, producing scaffolds with suitable features is crucial for guiding cells properly to develop the desired tissue-engineered vascular grafts for clinical applications. The main objective of this work is fabrication and characterization of tubular small-diameter ( < 6 mm) bilayered scaffolds for VTE. The scaffolds were prepared via mixing electrospinning approach of biodegradable poly(ε-caprolactone) (PCL) polymer – due to its favorable physicochemical properties – to mimic the natural environment-extracellular matrix. Firstly, tubular nanofibrous construct with inner diameter of 3, 4 or 5 mm was electrospun as inner layer, and secondly, microfibrous construct was electrospun as outer layer directly on the first produced inner layer. To improve the biological properties of PCL, a group of the electrospun scaffolds was immersed in type-1 collagen solution. The morphology and structure of the resulting fibrous scaffolds were investigated by scanning electron microscope. The electrospun nanofibrous inner layer contained fibers measuring 219±35 nm in diameter, while the electrospun microfibrous outer layer contained fibers measuring 1011 ± 150 nm. Furthermore, mechanical, thermal and physical tests were conducted with both electrospun bilayered scaffold types where revealed improved properties. Biological investigations using endothelial, smooth muscle and fibroblast cell line showed good biocompatibility of both tested electrospun scaffolds. Better attachment and proliferation were obviously found when cells were cultured on the scaffolds immersed with collagen due to increasing the hydrophilicity of the PCL. The easy, inexpensive and versatile electrospinning approach used in this work was able to successfully produce double layered tubular elastic structures containing both nanofibers and microfibers to imitate the native vascular structure. The PCL – as a suitable and approved biomaterial for many biomedical and tissue engineering applications – can ensure favorable mechanical properties of scaffolds used for VTE. The VTE approach using electrospun bilayered scaffolds offers optimal solutions and holds significant promises for treatment of many cardiovascular diseases.

Keywords: electrospinning, poly(ε-caprolactone) (PCL), tissue-engineered vascular graft, tubular bilayered scaffolds, vascular cells

Procedia PDF Downloads 295
3775 The Biology of Persister Cells and Antibiotic Resistance

Authors: Zikora K. G. Anyaegbunam, Annabel A. Nnawuihe, Ngozi J. Anyaegbunam, Emmanuel A. Eze

Abstract:

The discovery and production of new antibiotics is unavoidable in the fight against drug-resistant bacteria. However, this is only part of the problem; we have never really had medications that could completely eradicate an infection. All pathogens create a limited number of dormant persister cells that are resistant to antibiotic treatment. When the concentration of antibiotics decreases, surviving persisters repopulate the population, resulting in a recurrent chronic infection. Bacterial populations have an alternative survival strategy to withstand harsh conditions or antibiotic exposure, in addition to the well-known methods of antibiotic resistance and biofilm formation. Persister cells are a limited subset of transiently antibiotic-tolerant phenotypic variations capable of surviving high-dose antibiotic therapy. Persisters that flip back to a normal phenotype can restart growth when antibiotic pressure drops, assuring the bacterial population's survival. Persister cells have been found in every major pathogen, and they play a role in antibiotic tolerance in biofilms as well as the recalcitrance of chronic infections. Persister cells has been implicated to play a role in the establishment of antibiotic resistance, according to growing research. Thusthe need to basically elucidate the biology of persisters and how they are linked to antibiotic resistance, and as well it's link to diseases.

Keywords: persister cells, phenotypic variations, repopulation, mobile genetic transfers, antibiotic resistance

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3774 Design and Construction of a Device to Facilitate the Stretching of a Plantiflexors Muscles in the Therapy of Rehabilitation for Patients with Spastic Hemiplegia

Authors: Nathalia Andrea Calderon Lesmes, Eduardo Barragan Parada, Diego Fernando Villegas Bermudez

Abstract:

Spasticity in the plantiflexor muscles as a product of stroke (CVA-Cerebrovascular accident) restricts the mobility and independence of the affected people. Commonly, physiotherapists are in charge of manually performing the rehabilitation therapy known as Sustained Mechanical Stretching, rotating the affected foot of the patient in the sagittal plane. However, this causes a physical wear on the professional because it is a fatiguing movement. In this article, a mechanical device is developed to implement this rehabilitation therapy more efficiently. The device consists of a worm-crown mechanism that is driven by a crank to gradually rotate a platform in the sagittal plane of the affected foot, in order to achieve dorsiflexion. The device has a range of sagittal rotation up to 150° and has velcro located on the footplate that secures the foot. The design of this device was modeled by using CAD software and was checked structurally with a general purpose finite element software to be sure that the device is safe for human use. As a measurement system, a goniometer is used in the lateral part of the device and load cells are used to measure the force in order to determine the opposing torque exerted by the muscle. Load cells sensitivity is 1.8 ± 0.002 and has a repeatability of 0.03. Validation of the effectiveness of the device is measured by reducing the opposition torque and increasing mobility for a given patient. In this way, with a more efficient therapy, an improvement in the recovery of the patient's mobility and therefore in their quality of life can be achieved.

Keywords: biomechanics, mechanical device, plantiflexor muscles, rehabilitation, spastic hemiplegia, sustained mechanical stretching

Procedia PDF Downloads 167
3773 Determination of Antibiotic Residues in Carcasses of Cows Slaughtered in Amol City by Four-Plate-Test Method

Authors: Arezou Ghadi, Nasrollah Vahedi, Azam Sinkakarimi

Abstract:

For determination of antibiotic residues in slaughtered cow carcasses of Amol city in Iran, sampling has done from 100 heads of cow. For this purpose, the microbiological F.P.T (Four-Plate Test) method was used. Basis of this method, a clear zone is creating around the leachate on the plate that already has cultured a uniform layer of under test bacteria on agar plate. In this study from 100 heads of cow carcasses, at least 75 cases (75%) in one of the tested organs (muscle-liver-kidney) have been antibiotic residues. Also, it has been found that kidney have the most positive cases (60%) than other organs (liver and muscle), then the liver (58%) and finally are muscles (51%).

Keywords: antibiotic residues, agar plate test, cow carcass

Procedia PDF Downloads 457
3772 Typical Emulsions as Probiotic Food Carrier: Effect of Cells Position on Its Viability

Authors: Mengfan Li, Filip Van Bockstaele, Wenyong Lou, Frank Devlighere

Abstract:

The development of probiotics-encapsulated emulsions that maintain the viability of probiotics during processing, storage and human gastrointestinal (GI) tract environment receives great scientific and commercial interest. In this study, typical W/O and O/W emulsions with and without oil gelation were used to encapsulate L. plantarum. The effects of emulsion types on the viability of L. plantarum during storage and GI tract were investigated. Besides, the position of L. plantarum in emulsion system and its number of viable cells when threating by adverse environment was correlated in order to figure out which type of emulsion is more suitable as food carrier for probiotics encapsulation and protection. As a result, probiotics tend to migrate from oil to water phase due to the natural hydrophilicity; however, it’s harmful for cells viability when surrounding by water for a long time. Oil gelation in emulsions is one of the promising strategies for inhibiting the cells mobility and decreasing the contact with adverse factors (e.g., water, exogenous enzymes and gastric acid), thus enhancing the number of viable cells that enough to exert its beneficial effects in host.

Keywords: emulsion, gelation, encapsulation, probiotics

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3771 The Stability and Performances of Terminalia Catappa L. Dye-Sensitized Solar Cell

Authors: A. O. Boyo, A. T. Akinwunmi

Abstract:

The effect of extracting solvent and adjustment of pHs on the stability of Terminalia catappa L. dye-sensitized solar cell was investigated. We introduced ZnO as an alternative to TiO2 in the dye sensitized solar cells (DSSCs) due to its band gap similar to TiO2, higher electron mobility, and flexible procedures of preparations. Dye-sensitized solar cells (DSSCs) based on Terminalia catappa L. was extracted in water (A), ethanol (B) and the mixture of ethanol and water in the ratio 1:1by volume (C). The best performance Solar cells sensitized was from extracts A and achieved up to Jsc 1.51 mAcm−2, Voc 0.75V, FF 0.88 and η 0.63%. We notice that as pHs decreases there is the increase in DSSC efficiency. There is Long period stability in efficiency of the cells prepared using A than in C and a fair stability in efficiency of B cell. The results obtained with extracts B and C confirmed that Ethanol with water could not be considered as a suitable solvent for the extraction of natural dye.

Keywords: zinc oxide, dye-sensitized solar cell, terminalia catappa L., TiO2

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3770 Effect of Exercise Training on Body Composition and Metabolic Profile in Older Adults during Cancer Treatment

Authors: Adeline Fontvieille, Hugo Parent-Roberge, Marie-France Langlois, Tamas Fulop, Michel Pavic, Eleonor Riesco

Abstract:

Introduction: Total lean body mass is reduced during cancer treatment. This loss is called cancer cachexia and is accompanied by a progressive loss of fat mass. In older adults, these body composition changes can have a larger impact on metabolic health, physical autonomy, and cancer survival. Although currently untreatable, exercise training could reduce these effects. Hence, the objective of this pilot study is to investigate if 12 weeks of exercise training during cancer treatment can mitigate the loss of muscle mass and fat mass in older adults. Methods: A total of 40 older adults (65-80 years) with an ongoing treatment for a curable cancer are currently recruited and randomised in two groups: 1) Combined training (EX, n=20) and 2) Control group (CON, n=20). All variables are measured before and after 12 weeks of intervention: Anthropometry (weight, height, body mass index), body composition (total fat mass, visceral adipose tissue, total and appendicular muscle mass; DXA), metabolic profile (HDL-C and LDL-C, triglycerides, glucose and insulin levels). Results: Preliminary analyses revealed no impact of exercise training on appendicular muscle mass (p=0,31) and fat mass (p=0,31). Furthermore, total body weight, waist circumference, HDL-cholesterol, LDL-cholesterol, glucose and insulin levels remained unchanged (all p ≥ 0.79) after 12 weeks of training. However, statistical analyses revealed that triglyceride levels slightly increased (p=0.03), irrespective of the group. Conclusion: Preliminary analyses did not reveal any impact of aerobic and resistance exercise training on body composition in oncogeriatric patients. Furthermore, exercise training seems not efficient to prevent the cancer treatment-related triglyceride levels increase.

Keywords: muscle mass, fat mass, metabolic profile, combined training, aging, cancer

Procedia PDF Downloads 364
3769 Inflammatory Changes in Postmenopausal Women including Th17 and Treg

Authors: Ae Ra Han, Seoung Eun Huh, Ji Yeon Kim, Joanne Kwak-Kim, Sung Ki Lee

Abstract:

Objective: Prevalence of osteoporosis, cardiovascular disorders, and Alzheimer's disease rapidly increase after menopause. Immune activation and inflammation are suggested as an important pathogenesis of these serious diseases. Several pro-inflammatory cytokines are increased in women with surgical or natural menopause. However, the little is known about IL-17 producing T cells and Foxp3+ regulatory T (Treg) cells in post-menopause. Methods: A total of 34 postmenopausal women, who had no active cardiovascular, endocrine and infectious disorders were recruited as study group and healthy premenopausal women participated as controls. Peripheral blood mononuclear cells were isolated. Immuno-morphologic (CD3, CD4, CD8, CD19, CD56/CD16), intracellular cytokine (TNF-alpha, IFN-gamma, IL-10, IL-17), and Treg cell (Foxp3) studies were carried out using flow cytometry. The proportion of peripheral lymphocytes, including IL-17 producing and Foxp3+ Treg cells immune cell in each group were statistically analyzed. Results: The proportion of NK cells was significantly increased in menopausal women as compared to that of controls (P=.005). The ratios of TNF-alpha/IL-10 producing CD3+CD4+ T cells were increased in postmenopausal women. CD3+IL-17+ T cell level was higher in postmenopausal women and CD4+ Foxp3+ Treg cells was lower than that of controls. The ratios of CD3+IL-17+ T cell to CD3+Foxp3+ and to CD4+Foxp3+ Treg cells were significantly increased in postmenopausal women (P=.001). Conclusions: We found enhanced innate immunity and Th1- and Th17-mediated adaptive immunity in postmenopausal women. This may explain increasing prevalence of chronic inflammatory diseases after menopause. Further studies are needed to elucidate what factors contribute to this inflammatory shift in the postmenopause.

Keywords: inflammation, immune cell, menopause, Th17, regulatory T cell

Procedia PDF Downloads 323
3768 Acoustic Radiation Pressure Detaches Myoblast from Culture Substrate by Assistance of Serum-Free Medium

Authors: Yuta Kurashina, Chikahiro Imashiro, Kiyoshi Ohnuma, Kenjiro Takemura

Abstract:

Research objectives and goals: To realize clinical applications of regenerative medicine, a mass cell culture is highly required. In a conventional cell culture, trypsinization was employed for cell detachment. However, trypsinization causes proliferation decrease due to injury of cell membrane. In order to detach cells using an enzyme-free method, therefore, this study proposes a novel cell detachment method capable of detaching adherent cells using acoustic radiation pressure exposed to the dish by the assistance of serum-free medium with ITS liquid medium supplement. Methods used In order to generate acoustic radiation pressure, a piezoelectric ceramic plate was glued on a glass plate to configure an ultrasonic transducer. The glass plate and a chamber wall compose a chamber in which a culture dish is placed in glycerol. Glycerol transmits acoustic radiation pressure to adhered cells on the culture dish. To excite a resonance vibration of transducer, AC signal with 29-31 kHz (swept) and 150, 300, and 450 V was input to the transducer for 5 min. As a pretreatment to reduce cell adhesivity, serum-free medium with ITS liquid medium supplement was spread to the culture dish before exposed to acoustic radiation pressure. To evaluate the proposed cell detachment method, C2C12 myoblast cells (8.0 × 104 cells) were cultured on a ø35 culture dish for 48 hr, and then the medium was replaced with the serum-free medium with ITS liquid medium supplement for 24 hr. We replaced the medium with phosphate buffered saline and incubated cells for 10 min. After that, cells were exposed to the acoustic radiation pressure for 5 min. We also collected cells by using trypsinization as control. Cells collected by the proposed method and trypsinization were respectively reseeded in ø60 culture dishes and cultured for 24 hr. Then, the number of proliferated cells was counted. Results achieved: By a phase contrast microscope imaging, shrink of lamellipodia was observed before exposed to acoustic radiation pressure, and no cells remained on the culture dish after the exposed of acoustic radiation pressure. This result suggests that serum-free medium with ITS liquid inhibits adhesivity of cells and acoustic radiation pressure detaches cells from the dish. Moreover, the number of proliferated cells 24 hr after collected by the proposed method with 150 and 300 V is the same or more than that by trypsinization, i.e., cells were proliferated 15% higher with the proposed method using acoustic radiation pressure than with the traditional cell collecting method of trypsinization. These results proved that cells were able to be collected by using the appropriate exposure of acoustic radiation pressure. Conclusions: This study proposed a cell detachment method using acoustic radiation pressure by the assistance of serum-free medium. The proposed method provides an enzyme-free cell detachment method so that it may be used in future clinical applications instead of trypsinization.

Keywords: acoustic radiation pressure, cell detachment, enzyme free, ultrasonic transducer

Procedia PDF Downloads 254
3767 Deciphering Tumor Stroma Interactions in Retinoblastoma

Authors: Rajeswari Raguraman, Sowmya Parameswaran, Krishnakumar Subramanian, Jagat Kanwar, Rupinder Kanwar

Abstract:

Background: Tumor microenvironment has been implicated in several cancers to regulate cell growth, invasion and metastasis culminating in outcome of therapy. Tumor stroma consists of multiple cell types that are in constant cross-talk with the tumor cells to favour a pro-tumorigenic environment. Not much is known about the existence of tumor microenvironment in the pediatric intraocular malignancy, Retinoblastoma (RB). In the present study, we aim to understand the multiple stromal cellular subtypes and tumor stromal interactions expressed in RB tumors. Materials and Methods: Immunohistochemistry for stromal cell markers CD31, CD68, alpha-smooth muscle (α-SMA), vimentin and glial fibrillary acidic protein (GFAP) was performed on formalin fixed paraffin embedded tissues sections of RB (n=12). The differential expression of stromal target molecules; fibroblast activation protein (FAP), tenascin-C (TNC), osteopontin (SPP1), bone marrow stromal antigen 2 (BST2), stromal derived factor 2 and 4 (SDF2 and SDF4) in primary RB tumors (n=20) and normal retina (n=5) was studied by quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) and Western blotting. The differential expression was correlated with the histopathological features of RB. The interaction between RB cell lines (Weri-Rb-1, NCC-RbC-51) and Bone marrow stromal cells (BMSC) was also studied using direct co-culture and indirect co-culture methods. The functional effect of the co-culture methods on the RB cells was evaluated by invasion and proliferation assays. Global gene expression was studied by using Affymetrix 3’ IVT microarray. Pathway prediction was performed using KEGG and the key molecules were validated using qRT-PCR. Results: The immunohistochemistry revealed the presence of several stromal cell types such as endothelial cells (CD31+;Vim+/-); macrophages (CD68+;Vim+/-); Fibroblasts (Vim+; CD31-;CD68- );myofibroblasts (α-SMA+/ Vim+) and invading retinal astrocytes/ differentiated retinal glia (GFAP+; Vim+). A characteristic distribution of these stromal cell types was observed in the tumor microenvironment, with endothelial cells predominantly seen in blood vessels and macrophages near actively proliferating tumor or necrotic areas. Retinal astrocytes and glia were predominant near the optic nerve regions in invasive tumors with sparse distribution in tumor foci. Fibroblasts were widely distributed with rare evidence of myofibroblasts in the tumor. Both gene and protein expression revealed statistically significant (P<0.05) up-regulation of FAP, TNC and BST2 in primary RB tumors compared to the normal retina. Co-culture of BMSC with RB cells promoted invasion and proliferation of RB cells in direct and indirect contact methods respectively. Direct co-culture of RB cell lines with BMSC resulted in gene expression changes in ECM-receptor interaction, focal adhesion, IL-8 and TGF-β signaling pathways associated with cancer. In contrast, various metabolic pathways such a glucose, fructose and amino acid metabolism were significantly altered under the indirect co-culture condition. Conclusion: The study suggests that the close interaction between RB cells and the stroma might be involved in RB tumor invasion and progression which is likely to be mediated by ECM-receptor interactions and secretory factors. Targeting the tumor stroma would be an attractive option for redesigning treatment strategies for RB.

Keywords: gene expression profiles, retinoblastoma, stromal cells, tumor microenvironment

Procedia PDF Downloads 385