Search results for: multi-well 3d cell culture

Authors: E. M. Hassan, A. L. Kalamkarov

Abstract:

Two micromechanical models for 3D smart composite with embedded periodic or nearly periodic network of generally orthotropic reinforcements and actuators are developed and applied to cubic structures with unidirectional orientation of constituents. Analytical formulas for the effective piezothermoelastic coefficients are derived using the Asymptotic Homogenization Method (AHM). Finite Element Analysis (FEA) is subsequently developed and used to examine the aforementioned periodic 3D network reinforced smart structures. The deformation responses from the FE simulations are used to extract effective coefficients. The results from both techniques are compared. This work considers piezoelectric materials that respond linearly to changes in electric field, electric displacement, mechanical stress and strain and thermal effects. This combination of electric fields and thermo-mechanical response in smart composite structures is characterized by piezoelectric and thermal expansion coefficients. The problem is represented by unit-cell and the models are developed using the AHM and the FEA to determine the effective piezoelectric and thermal expansion coefficients. Each unit cell contains a number of orthotropic inclusions in the form of structural reinforcements and actuators. Using matrix representation of the coupled response of the unit cell, the effective piezoelectric and thermal expansion coefficients are calculated and compared with results of the asymptotic homogenization method. A very good agreement is shown between these two approaches.

Keywords: asymptotic homogenization method, finite element analysis, effective piezothermoelastic coefficients, 3D smart network composite structures

Procedia PDF Downloads 398

Authors: Raluca-Ioana Antonescu

Abstract:

The field research of the paper stands at the intersection between Linguistics and Sociology, while the problem of the research is the importance of language in the modernization process and in a globalized society. The research objective is to prove that language is a stimulant for modernity, while it defines the tradition and the culture of a specific society. In order to examine the linguistic change of the Korean language due to the modernity and globalization, the paper tries to answer one main question, What are the changes the Korean language underwent from a traditional version of Korean, towards one influenced by modernity?, and two secondary questions, How are explored in specialized literature the relations between globalization (and modernity) and culture (focusing on language)? and What influences the Korean language? For the purpose of answering the research questions, the paper has the main premise that due to modernity and globalization, the Korean language changed its discourse construction, and two secondary hypothesis, first is that in literature there are not much explored the relations between culture and modernity focusing on the language discourse construction, but more about identity issue and commodification problems, and the second hypothesis is that the Korean language is influenced by traditional values (like Confucianism) while receiving influence also of globalization process (especially from English language). In terms of methodology, the paper will analyze the two main influences upon the Korean language, referring to traditionalism (being defined as the influence of Confucianism) and modernism (as the influence of other countries’ language and culture), and how the Korean language it was constructed and modified due to these two elements. The paper will analyze at what level (grammatical, lexical, etc.) the traditionalism help at the construction of the Korean language, and what are the changes at each level that modernism brought along. As for the results of this research, the influence of modernism changed both lexically and grammatically the Korean language. In 60 years the increase of English influence is astonishing, and this paper shows the main changes the Korean language underwent, like the loanwords (Konglish), but also the reduction of the speech levels and the ease of the register variation use. Therefore the grammatical influence of modernity and globalization could be seen at the reduction of the speech level and register variation, while the lexical change comes with the influence of English language especially, where about 10% of the Korean vocabulary is considered to be loanwords. Also the paper presents the interrelation between traditionalism and modernity, with the example of Konglish, but not only (we can consider also the Korean greetings which are translated by Koreans when they speak in other languages, bringing their cultural characteristics in English discourse construction), which makes the Koreans global, since they speak in an international language, but still local since they cannot get rid completely of their culture.

Keywords: Confucianism, globalization, language and linguistic change, modernism, traditionalism

Procedia PDF Downloads 196

Authors: Leander Van Neste, Kirk Wojno

Abstract:

The innate immune system reacts to foreign insult in several unique ways, one of which is phagocytosis of perceived threats such as cancer, bacteria, and viruses. The goal of this study was to look for evidence of phagocytosed RNA from tumor cells in circulating monocytes. While all monocytes possess phagocytic capabilities, the non-classical CD14+/FCGR3A+ monocytes and the intermediate CD14++/FCGR3A+ monocytes most actively remove threatening ‘external’ cellular materials. Purified CD14-positive monocyte samples from fourteen patients recently diagnosed with clinically localized prostate cancer (PCa) were investigated by single-cell RNA sequencing using the 10X Genomics protocol followed by paired-end sequencing on Illumina’s NovaSeq. Similarly, samples were processed and used as controls, i.e., one patient underwent biopsy but was found not to harbor prostate cancer (benign), three young, healthy men, and three men previously diagnosed with prostate cancer that recently underwent (curative) radical prostatectomy (post-RP). Sequencing data were mapped using 10X Genomics’ CellRanger software and viable cells were subsequently identified using CellBender, removing technical artifacts such as doublets and non-cellular RNA. Next, data analysis was performed in R, using the Seurat package. Because the main goal was to identify differences between PCa patients and ‘control’ patients, rather than exploring differences between individual subjects, the individual Seurat objects of all 21 patients were merged into one Seurat object per Seurat’s recommendation. Finally, the single-cell dataset was normalized as a whole prior to further analysis. Cell identity was assessed using the SingleR and cell dex packages. The Monaco Immune Data was selected as the reference dataset, consisting of bulk RNA-seq data of sorted human immune cells. The Monaco classification was supplemented with normalized PCa data obtained from The Cancer Genome Atlas (TCGA), which consists of bulk RNA sequencing data from 499 prostate tumor tissues (including 1 metastatic) and 52 (adjacent) normal prostate tissues. SingleR was subsequently run on the combined immune cell and PCa datasets. As expected, the vast majority of cells were labeled as having a monocytic origin (~90%), with the most noticeable difference being the larger number of intermediate monocytes in the PCa patients (13.6% versus 7.1%; p<.001). In men harboring PCa, 0.60% of all purified monocytes were classified as harboring PCa signals when the TCGA data were included. This was 3-fold, 7.5-fold, and 4-fold higher compared to post-RP, benign, and young men, respectively (all p<.001). In addition, with 7.91%, the number of unclassified cells, i.e., cells with pruned labels due to high uncertainty of the assigned label, was also highest in men with PCa, compared to 3.51%, 2.67%, and 5.51% of cells in post-RP, benign, and young men, respectively (all p<.001). It can be postulated that actively phagocytosing cells are hardest to classify due to their dual immune cell and foreign cell nature. Hence, the higher number of unclassified cells and intermediate monocytes in PCa patients might reflect higher phagocytic activity due to tumor burden. This also illustrates that small numbers (~1%) of circulating peripheral blood monocytes that have interacted with tumor cells might still possess detectable phagocytosed tumor RNA.

Keywords: circulating monocytes, phagocytic cells, prostate cancer, tumor immune response

Procedia PDF Downloads 159

Authors: Fariyal Mir

Abstract:

The largest minority group over Pakistan lives in the farthest region of KPK, Chitral, which enclosed all its people in one closet despite their diversity. Chitral is considered a tourist hub and piece of paradise on earth. The major attraction of Chitral is the Kalash valley, which is also known as the homeland of kafir or 'weavers of the black robes'. Kalash people are counted as the primitive pagan tribe who practice culture, beliefs, and a lifestyle which dated beyond and distinct from the rest of the region. Their religious belief is known to be as 'animism', which means that objects, places, and creatures all possess a distinct spiritual essence. They also believe that nature plays a highly significant and spiritual role in their daily life. As they have their different festivals like (a) Chilam Josh: celebrated in May to welcome spring, (b) Uchaw: which is observed in late August to ensure good crops of wheat, (c) Chanmos: which is being observed in December for more than two weeks, grand festival and is celebrated to welcome New Year. Their concept of purity and impurity is also very traditional and part of their religious belief as well. They used to call the man as pure 'Oshniru' (pure, holy) and woman as impure 'Chetu' (dirty, polluted, or contaminated). This study is based on ethnomethodology, which focuses on the way people make sense of their daily life. Their unique belief system connects them with the descendants of the armies of Alexander the Great, who called the Hindu Kush 'Parapamisus', meaning mountains over which eagles can fly. With time, their cultural system was molded into many ways, and the same is going on with the true beauty of Chitral in the form of conversion to Islam and adaptation of modern lifestyle. Ethnomethodology also supports this, that people are always good in their original form so they can be called the true representatives of their origin and cultural identity. It also argues that human society is based on the method of achieving and displaying not on the imposition of others' order and laws. So, everybody is obliged to respect and not to harm the beliefs and culture of all other human fellows. Some historians believe that these Kalash people are from Afghanistan and their origin is Saim which is known as Thailand, but those Kalash tribe from Afghanistan were forced to embrace Islam and only Pakistan keeps these people who own unique cultural values, ornaments and entire culture. Kalasha culture has been listed by UNESCO for consideration as World Heritage. It is necessary to save these people from the cultural invasion caused by modernity, forced religious conversion to maintain the pluralistic diversity of our country.

Keywords: Chitral, cultural identity, ethnomethodology, Kalasha culture

Procedia PDF Downloads 146

Authors: Marinna Madrid, Nabiha Saklayen, Marinus Huber, Nicolas Vogel, Christos Boutopoulos, Michel Meunier, Eric Mazur

Abstract:

Delivering material into cells is important for a diverse range of biological applications, including gene therapy, cellular engineering and imaging. We present a plasmonic substrate for delivering membrane-impermeable material into cells at high throughput and high efficiency while maintaining cell viability. The substrate fabrication is based on an affordable and fast colloidal self-assembly process. When illuminated with a femtosecond laser, the light interacts with the electrons at the surface of the metal substrate, creating localized surface plasmons that form bubbles via energy dissipation in the surrounding medium. These bubbles come into close contact with the cell membrane to form transient pores and enable entry of membrane-impermeable material via diffusion. We use fluorescence microscopy and flow cytometry to verify delivery of membrane-impermeable material into HeLa CCL-2 cells. We show delivery efficiency and cell viability data for a range of membrane-impermeable cargo, including dyes and biologically relevant material such as siRNA. We estimate the effective pore size by determining delivery efficiency for hard fluorescent spheres with diameters ranging from 20 nm to 2 um. To provide insight to the cell poration mechanism, we relate the poration data to pump-probe measurements of micro- and nano-bubble formation on the plasmonic substrate. Finally, we investigate substrate stability and reusability by using scanning electron microscopy (SEM) to inspect for damage on the substrate after laser treatment. SEM images show no visible damage. Our findings indicate that self-assembled plasmonic substrates are an affordable tool for high-throughput, high-efficiency delivery of material into mammalian cells.

Keywords: femtosecond laser, intracellular delivery, plasmonic, self-assembly

Procedia PDF Downloads 526

Authors: Anil Koklu, Amin Mansoorifar, Ali Beskok

Abstract:

Dielectric spectroscopy (DS) is a noninvasive, label free technique for a long term real-time measurements of the impedance spectra of biological cells. DS enables characterization of cellular dielectric properties such as membrane capacitance and cytoplasmic conductivity. We have developed a lab-on-a-chip device that uses an electro-activated microwells array for loading, DS measurements, and unloading of biological cells. We utilized from dielectrophoresis (DEP) to capture target cells inside the wells and release them after DS measurement. DEP is a label-free technique that exploits differences among dielectric properties of the particles. In detail, DEP is the motion of polarizable particles suspended in an ionic solution and subjected to a spatially non-uniform external electric field. To the best of our knowledge, this is the first microfluidic chip that combines DEP and DS to analyze biological cells using electro-activated wells. Device performance is tested using two different cell lines of prostate cancer cells (RV122, PC-3). Impedance measurements were conducted at 0.2 V in the 10 kHz to 40 MHz range with 6 s time resolution. An equivalent circuit model was developed to extract the cell membrane capacitance and cell cytoplasmic conductivity from the impedance spectra. We report the time course of the variations in dielectric properties of PC-3 and RV122 cells suspended in low conductivity medium (LCB), which enhances dielectrophoretic and impedance responses, and their response to sudden pH change from a pH of 7.3 to a pH of 5.8. It is shown that microfluidic chip allowed online measurements of dielectric properties of prostate cancer cells and the assessment of the cellular level variations under external stimuli such as different buffer conductivity and pH. Based on these data, we intend to deploy the current device for single cell measurements by fabricating separately addressable N × N electrode platforms. Such a device will allow time-dependent dielectric response measurements for individual cells with the ability of selectively releasing them using negative-DEP and pressure driven flow.

Keywords: microfluidic, microfabrication, lab on a chip, AC electrokinetics, dielectric spectroscopy

Procedia PDF Downloads 148

Authors: Sajeli A. Begum, Ameer Basha, Kirti Hira, Rukaiyya Khan

Abstract:

Cyclic dipeptides are simple diketopiperazine derivatives being investigated by several scientists for their biological effects which include anticancer, antimicrobial, haematological, anticonvulsant, immunomodulatory effect, etc. They are potentially active microbial metabolites having been synthesized too, for developing into drug candidates. Cultures of Pseudomonas species have earlier been reported to produce cyclic dipeptides, helping in quorum sensing signals and bacterial–host colonization phenomena during infections, causing cell anti-proliferation and immunosuppression. Fluorescing Pseudomonas species have been identified to secrete lipid derivatives, peptides, pyrroles, phenazines, indoles, aminoacids, pterines, pseudomonic acids and some antibiotics. In the present work, results of investigation on the cyclic dipeptide metabolites secreted by the culture broth of Pseudomonas species as potent pro-inflammatory cytokine inhibitors are discussed. The bacterial strain was isolated from the rhizospheric soil of groundnut crop and identified as Pseudomonas aeruginosa by 16S rDNA sequence (GenBank Accession No. KT625586). Culture broth of this strain was prepared by inoculating into King’s B broth and incubating at 30 ºC for 7 days. The ethyl acetate extract of culture broth was prepared and lyophilized to get a dry residue (EEPA). Lipopolysaccharide (LPS)-induced ELISA assay proved the inhibition of tumor necrosis factor-alpha (TNF-α) secretion in culture supernatant of RAW 264.7 cells by EEPA (IC50 38.8 μg/mL). The effect of oral administration of EEPA on plasma TNF-α level in rats was tested by ELISA kit. The LPS mediated plasma TNF-α level was reduced to 45% with 125 mg/kg dose of EEPA. Isolation of the chemical constituents of EEPA through column chromatography yielded ten cyclic dipeptides, which were characterized using nuclear magnetic resonance and mass spectroscopic techniques. These cyclic dipeptides are biosynthesized in microorganisms by multifunctional assembly of non-ribosomal peptide synthases and cyclic dipeptide synthase. Cyclo (Gly-L-Pro) was found to be more potentially (IC50 value 4.5 μg/mL) inhibiting TNF-α production followed by cyclo (trans-4-hydroxy-L-Pro-L-Phe) (IC50 value 14.2 μg/mL) and the effect was equal to that of standard immunosuppressant drug, prednisolone. Further, the effect was analyzed by determining mRNA expression of TNF-α in LPS-stimulated RAW 264.7 macrophages using quantitative real-time reverse transcription polymerase chain reaction. EEPA and isolated cyclic dipeptides demonstrated diminution of TNF-α mRNA expression levels in a dose-dependent manner under the tested conditions. Also, they were found to control the expression of other pro-inflammatory cytokines like IL-1β and IL-6, when tested through their mRNA expression levels in LPS-stimulated RAW 264.7 macrophages under LPS-stimulated conditions. In addition, significant inhibition effect was found on Nitric oxide production. Further all the compounds exhibited weak toxicity to LPS-induced RAW 264.7 cells. Thus the outcome of the study disclosed the effectiveness of EEPA and the isolated cyclic dipeptides in down-regulating key cytokines involved in pathophysiology of autoimmune diseases.In another study led by the investigators, microbial cyclic dipeptides were found to exhibit excellent antimicrobial effect against Fusarium moniliforme which is an important causative agent of Sorghum grain mold disease. Thus, cyclic dipeptides are emerging small molecular drug candidates for various autoimmune diseases.

Keywords: cyclic dipeptides, cytokines, Fusarium moniliforme, Pseudomonas, TNF-alpha

Procedia PDF Downloads 208

Authors: Denis Mustafov, Emmanouil Karteris, Maria Braoudaki

Abstract:

Glioblastoma multiform (GBM) is a heterogenous primary brain tumour that kills most affected patients. To the authors best knowledge, despite all research efforts there is no early diagnostic biomarker for GBM. MicroRNAs (miRNAs) are short non-coding RNA molecules which are deregulated in many cancers. The aim of this research was to determine miRNAs with a diagnostic impact and to potentially identify promising therapeutic targets for glioblastoma multiform. In silico analysis was performed to identify deregulated miRNAs with diagnostic relevance for glioblastoma. The expression profiles of the chosen miRNAs were then validated in vitro in the human glioblastoma cell lines A172 and U-87MG. Briefly, RNA extraction was carried out using the Trizol method, whilst miRNA extraction was performed using the mirVANA miRNA isolation kit. Quantitative Real-Time Polymerase Chain Reaction was performed to verify their expression. The presence of five target proteins within the A172 cell line was evaluated by Western blotting. The expression of the CORO1C protein within 32 GBM cases was examined via immunohistochemistry. The miRNAs identified in silico included miR-21-5p, miR-34a and miR-128a. These miRNAs were shown to target deregulated GBM genes, such as CDK6, E2F3, BMI1, JAG1, and CORO1C. miR-34a and miR-128a showed low expression profiles in comparison to a control miR-RNU-44 in both GBM cell lines suggesting tumour suppressor properties. Opposing, miR-21-5p demonstrated greater expression indicating that it could potentially function as an oncomiR. Western blotting revealed expression of all five proteins within the A172 cell line. In silico analysis also suggested that CORO1C is a target of miR-128a and miR-34a. Immunohistochemistry demonstrated that 75% of the GBM cases showed moderate to high expression of CORO1C protein. Greater understanding of the deregulated expression of miR-128a and the upregulation of CORO1C in GBM could potentially lead to the identification of a promising diagnostic biomarker signature for glioblastomas.

Keywords: non-coding RNAs, gene expression, brain tumours, immunohistochemistry

Procedia PDF Downloads 83

Authors: Uftah Ali M. Shushni, Viola Stuppec, Ulrike Lindequist

Abstract:

Because of the evolving resistance of microorganisms to existing antibiotics, there is an increasing need for new antibiotics not only in human but also in veterinary medicine. As part of our ongoing work on the secondary metabolites produced by marine fungi, the organic extract of the culture filtrate of an Ascomycetous fungus, which was found on driftwood collected from the coast of the Greifswalder Bodden, Baltic Sea, Germany displayed antimicrobial activity against some fish and human pathogenic bacteria. Bioactivity-guided column chromatographic separation led to the isolation of 6-Deoxybostrycoidin. The structure was determined from the interpretation of spectroscopic data (UV, MS, and NMR). 6-Deoxybostrycoidin exhibited in vitro activity against Bacillus subtilis, Staphylococcus aureus and Flexibacter maritimus with minimal inhibitory concentrations of 25, 12.5 and 12.5 μg/ml respectively.

Keywords: marine fungi, fish pathogenic bacteria, microorganism, medicine

Procedia PDF Downloads 527

Authors: Hala M. El-hanbuli, Mohammed F. Darweesh

Abstract:

The incidence rates of any tumor vary hugely with geographical location. Basal Cell Carcinoma (BCC) is one of the most common skin cancer that has many histopathologic subtypes. Objective: The aim was to study the histopathological features of BCC cases that were received in the Pathology Department, Kasr El-Aini hospital, Cairo University, Egypt during the period from Jan 2004 to Dec 2013 and to evaluate the clinical characters through the patient data available in the request sheets. Methods: Slides and data of BCC cases were collected from the archives of the pathology department, Kasr El-Aini hospital. Revision of all available slides and histological classification of BCC according to WHO (2006) was done. Results: A total number of 310 cases of BCC representing about 65% from the total number of malignant skin tumors examined during the 10-years duration in the department. The age ranged from 8 to 84 years, the mean age was (55.7 ± 15.5). Most of the patients (85%) were above the age of 40 years. There was a slight male predominance (55%). Ulcerated BCC was the most common gross picture (60%), followed by nodular lesion (30%) and finally the ulcerated nodule (10%). Most of the lesions situated in the high-risk sites (77%) where the nose was the most common site (35%) followed by the periocular area (22%), then periauricular (15%) and finally perioral (5%). No lesion was reported outside the head. The tumor size was less than 2 centimeters in 65% of cases, and from 2-5 centimeters in the lesions' greatest dimension in the rest of cases. Histopathological reclassification revealed that the nodular BCC was the most common (68%) followed by the pigmented nodular (18.75%). The histologic high-risk groups represented (7.5%) about half of them (3.75%) being basosquamous carcinoma. The total incidence for multiple BCC and 2nd primary was 12%. Recurrent BCC represented 8%. All of the recurrent lesions of BCC belonged to the histologic high-risk group. Conclusion: Basal Cell Carcinoma is the most common skin cancer in the 10-year survey. Histopathological diagnosis and classification of BCC cases are essential for the determination of the tumor type and its biological behavior.

Keywords: basal cell carcinoma, high risk, histopathological features, statistical analysis

Procedia PDF Downloads 148

Authors: Goodness Thandi Ntuli

Abstract:

Some cultural aspects in the African context have a tendency of uplifting women while some thrust them into the worst denigration scenarios; hence African Women Theologians refer to culture as a ‘double edged sword. Through socialization and internalization of social norms, some women become custodians of life, denying aspects of the culture that are against them and hand them down to the next generation. This indirectly contributes to the perpetuation of patriarchal tendencies wherein women themselves uphold and endorse such tendencies to their own detriment. One of the findings of the empirical research study conducted among the Zulu young women in the South African context was that, on the one hand, lobola (the bride-price) is one of the cultural practices that contribute a great deal in the vilification of women. On the other hand, a woman whose lobola has been paid is highly esteemed in the cultural context not only by society at large but also by the implicated woman who takes pride in it. Consequently, lobola becomes an ambiguous cultural practice. Thus from the postcolonial feminist perspective, this paper examines and critiques lobola practice while also disclosing and exposing its deep seated cultural reinforcement that is life denying to women. The paper elucidates the original lobola as a cultural practice before colonization and how it became commercialized during colonial times. With commercialization in the modern world, lobola has completely lost its preliminary meaning and ceased to be a life-giving cultural practice, particularly for women. It turned out to be the worst cultural practice that demeans women to the extent that it becomes suicidal to women dignity because, in marriage, they become objects or property to the men who purchased them. Women objectification in marriage does not only leave them culturally trapped in what was perceived to be a good practice, but it also leads to women abuse and gender based or domestic violence. The research has indicated that this kind of violence is escalating and has become so pervasive in the South African context that the country is rated as one of the capital cities of violence against women in the world. Therefore, this paper demonstrates how cultural practices at times indirectly contribute to this national scourge that needs to be condemned, disparaged and rejected. Women in the African context where such cultural activities are still viewed as a norm are in desperate need for true liberation from such ambiguous cultural practices that leave them in the margins in spite of the earned social status they might have achieved.

Keywords: african, ambiguity, critique, culture, feminist, lobola, postcolonial, society

Procedia PDF Downloads 190

Authors: Nathalie Quinto, Danielle Solancho

Abstract:

In 1986, after Marcos’ Martial Law of 1972, the Philippines revised its constitution for the fifth time, under the Aquino Administration. Extrajudicial violence was expected to be lessened, if not completely eradicated after this was passed. However, state-sponsored executions continued to persist even in the present time. There are currently identified policy gaps when it comes to extrajudicial cases, as there is no generally accepted definition of the term in the Philippines. In this paper, a triangulation method of historically published papers, key informant interviews, and focus group discussions of academics, scholars, and people who are involved in various cases found, was utilized for the methodology. This paper explores the establishment of a normalized system of state-sponsored executions in the country and why the state resorts to this kind of action. It found that due to a weak political, and social institution, a culture of extrajudicial executions was established.

Keywords: extrajudicial execution, human rights, justice, security

Procedia PDF Downloads 258

Authors: Sanjay Kumar Behera, Kanhu Charan Patra

Abstract:

A GIS-based method has been applied for the determination of soil erosion and sediment yield in a small watershed in Ong River basin, Odisha, India. The method involves spatial disintegration of the catchment into homogenous grid cells to capture the catchment heterogeneity. The gross soil erosion in each cell was calculated using Universal Soil Loss Equation (USLE) by carefully determining its various parameters. The concept of sediment delivery ratio is used to route surface erosion from each of the discretized cells to the catchment outlet. The process of sediment delivery from grid cells to the catchment outlet is represented by the topographical characteristics of the cells. The effect of DEM resolution on sediment yield is analyzed using two different resolutions of DEM. The spatial discretization of the catchment and derivation of the physical parameters related to erosion in the cell are performed through GIS techniques.

Keywords: DEM, GIS, sediment delivery ratio, sediment yield, soil erosion

Procedia PDF Downloads 444

Authors: Shima Fasahat

Abstract:

This research is an experimental research which was done about microbial fuel cells in order to study them for electricity generating and wastewater treatment. These days, it is very important to find new, clean and sustainable ways for energy supplying. Because of this reason there are many researchers around the world who are studying about new and sustainable energies. There are different ways to produce these kind of energies like: solar cells, wind turbines, geothermal energy, fuel cells and many other ways. Fuel cells have different types one of these types is microbial fuel cell. In this research, an MFC was built in order to study how it can be used for electricity generating and wastewater treatment. The microbial fuel cell which was used in this research is a reactor that has two tanks with a catalyst solution. The chemical reaction in microbial fuel cells is a redox reaction. The microbial fuel cell in this research is a two chamber MFC. Anode chamber is an anaerobic one (ABR reactor) and the other chamber is a cathode chamber. Anode chamber consists of stabilized sludge which is the source of microorganisms that do redox reaction. The main microorganisms here are: Propionibacterium and Clostridium. The electrodes of anode chamber are graphite pages. Cathode chamber consists of graphite page electrodes and catalysts like: O₂, KMnO₄ and C₆N₆FeK₄. The membrane which separates the chambers is Nafion117. The reason of choosing this membrane is explained in the complete paper. The main goal of this research is to generate electricity and treating wastewater. It was found that when you use electron receptor compounds like: O_2, MnO₄, C₆N₆FeK₄ the velocity of electron receiving speeds up and in a less time more current will be achieved. It was found that the best compounds for this purpose are compounds which have iron in their chemical formula. It is also important to pay attention to the amount of nutrients which enters to bacteria chamber. By adding extra nutrients in some cases the result will be reverse.  By using ABR the amount of chemical oxidation demand reduces per day till it arrives to a stable amount.

Keywords: anaerobic baffled reactor, bioenergy, electrode, energy efficient, microbial fuel cell, renewable chemicals, sustainable

Procedia PDF Downloads 223

Authors: Aditi Mittal, Nishit Gupta, Tina Dadu, Anil Handoo

Abstract:

Introduction: Hematopoietic stem cell transplantation (HSCT) is a curative treatment done for hematologic malignancies and other clinical conditions. Its main objective is to reconstitute the hematopoietic system of the recipient by administering an infusion of donor hematopoietic stem cells. Transplant engraftment is the first sign of bone marrow recovery. The main objective of this study is to assess immature platelet fraction (IPF) and immature reticulocyte fraction (IRF) as early indicators of post-hematopoietic stem cell transplant engraftment. Methods: Patients of all age groups and both genders undergoing both autologous and allogeneic transplants were included in the study. All the CBC samples were run on Mindray CAL-8000 (BC-6800 plus; Shenzhen, China) analyser and assessed for IPF and IRF. Neutrophil engraftment was defined as the first of three consecutive days with an ANC >0.5 x 109/L and platelet engraftment with a count >20 x 109/L. The cut-off values for IRF were calculated as 13.5% with a CV of 5% and for IPF was 19% with a CV of 12%. Results: The study sample comprised 200 patients, of whom 116 had undergone autologous HSCT, and 84 had undergone allogeneic HSCT. We observed that IRF anticipated the neutrophil recovery by an average of 5 days prior to IPF. Though there was no significant variation in IPF and IRF for the prediction of platelet recovery, IRF was preceded by 1 or 2 days to IPF in 25% of cases. Conclusions: Both IPF and IRF can be used as reliable parameters as predictors for post-transplant engraftment; however, IRF seems to be more reliable than IPF as a simple, inexpensive, and widely available tool for predicting marrow recovery several days before engraftment.

Keywords: transplantation, stem cells, reticulocyte, engraftment

Procedia PDF Downloads 85

Authors: Chantal Maille

Abstract:

This paper is about gender-based analysis plus (GBA+), an intersectional gender policy used in Canada to assess the impact of policies and programs for men and women from different origins. It looks at Canada’s political culture to explain the nature of its gender policies. GBA+ is defined as an analysis method that makes it possible to assess the eventual effects of policies, programs, services, and other initiatives on women and men of different backgrounds because it takes account of gender and other identity factors. The ‘plus’ in the name serves to emphasize that GBA+ goes beyond gender to include an examination of a wide range of other related identity factors, such as age, education, language, geography, culture, and income. The point of departure for GBA+ is that women and men are not homogeneous populations and gender is never the only factor in defining a person’s identity; rather, it interacts with factors such as ethnic origin, age, disabilities, where the person lives, and other aspects of individual and social identity. GBA+ takes account of these factors and thus challenges notions of similarity or homogeneity within populations of women and men. Comparative analysis based on sex and gender may serve as a gateway to studying a given question, but women, men, girls, and boys do not form homogeneous populations. In the 1990s, intersectionality emerged as a new feminist framework. The popularity of the notion of intersectionality corresponds to a time when, in hindsight, the damage done to minoritized groups by state disengagement policies in concert with global intensification of neoliberalism, and vice versa, can be measured. Although GBA+ constitutes a form of intersectionalization of GBA, it must be understood that the two frameworks do not spring from a similar logic. Intersectionality first emerged as a dynamic analysis of differences between women that was oriented toward change and social justice, whereas GBA is a technique developed by state feminists in a context of analyzing governmental policies and aiming to promote equality between men and women. It can nevertheless be assumed that there might be interest in such a policy and program analysis grid that is decentred from gender and offers enough flexibility to take account of a group of inequalities. In terms of methodology, the research is supported by a qualitative analysis of governmental documents about GBA+ in Canada. Research findings identify links between Canadian gender policies and its political culture. In Canada, diversity has been taken into account as an element at the basis of gendered analysis of public policies since 1995. The GBA+ adopted by the government of Canada conveys an opening to intersectionality and a sensitivity to multiculturalism. The Canadian Multiculturalism Act, adopted 1988, proposes to recognize the fact that multiculturalism is a fundamental characteristic of the Canadian identity and heritage and constitutes an invaluable resource for the future of the country. In conclusion, Canada’s distinct political culture can be associated with the specific nature of its gender policies.

Keywords: Canada, gender-based analysis, gender policies, political culture

Procedia PDF Downloads 220

Authors: Sendy Ghirardi, Pau Rausell Köster

Abstract:

The European Commission increasingly pushes cultural policies towards social outcomes and local and regional authorities also call for culture-driven strategies for local development and prosperity and therefore, the measurement of cultural participation becomes increasingly more significant for evidence-based policy-making processes. Cultural participation involves various kinds of social and economic spillovers that combine social and economic objectives of value creation, including social sustainability and respect for human values. Traditionally, from the economic perspective, cultural consumption is measured by the value of financial transactions in purchasing, subscribing to, or renting cultural equipment and content, addressing the market value of cultural products and services. The main sources of data are the household spending survey and merchandise trade survey, among others. However, what characterizes the cultural consumption is that it is linked with the hedonistic and affective dimension rather than the utilitarian one. In fact, nowadays, more and more attention is being paid to the social and psychological dimensions of culture. The aim of this work is to present a comprehensive approach to measure the impacts of cultural participation and cultural users’ behaviour, combining both socio-psychological and economic approaches. The model combines contingent evaluation techniques with the individual characteristic and perception analysis of the cultural experiences to evaluate the cognitive, aesthetic, emotive and social impacts of cultural participation. To investigate the comprehensive approach to measure the impact of the cultural events on individuals, the research has been designed on the basis of prior theoretical development. A deep literature methodology has been done to develop the theoretical model applied to the web platform to measure the impacts of cultural experience on individuals. The developed framework aims to become a democratic tool for evaluating the services that cultural or policy institutions can adopt through the use of an interacting platform that produces big data benefiting academia, cultural management and policies. The Au Culture is a prototype based on an application that can be used on mobile phones or any other digital platform. The development of the AU Culture Platform has been funded by the Valencian Innovation Agency (Government of the Region of Valencia) and it is part of the Horizon 2020 project MESOC.

Keywords: comprehensive approach, cultural participation, economic dimension, socio-psychological dimension

Procedia PDF Downloads 111

Authors: H. M. Avanesyan

Abstract:

Economists have been discussing the importance of intangible assets for the success of organization for many years. The term intellectual capital was popularized in the 1990s by Thomas Stewart. “Intellectual capital is the knowledge, applied experience, enterprise processes and technology customer relationship and professional skills which are valuable assets to an organization.” Human capital – includes employee brainpower, competence, skills, experience and knowledge. Customer capital – includes relations and networks with partners, suppliers, distributors, and customers. The objective of the article is to assess one of the key components of organizational culture – organizational values. The focus of the survey was on assessing how intellectual capital presented in these values of the organization. In the conclusion section the article refers to underestimation of intellectual capital by the organization management and the various possible negative effects of the latter.

Keywords: human capital, intellectual capital, organizational culture, management, social identity, organization

Procedia PDF Downloads 459

Authors: Vishnu Varthan Vaithiyalingam Jagannathan, Lakshmi Karunanidhi Santhanalakshmi, Srividya Ammayappan Rajam

Abstract:

Formononetin is the O-Methoxy Flavonol that has many pharmacological activities, which belongs to the flavonoid family. In the current study, activity of this molecule was evaluated in lung cancer cell lines. In general, flavonoids possess certain anticancer mechanism. Being a flavonoid subfamily, this molecule was subjected to evaluate cytotoxicity assay by MTT ((3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide)) stain, mode of cell death assay stained by acridine orange and ethidium bromide and Evaluation of Apoptosis pathway (extrinsic or intrinsic) by Caspase 3/7 stain and Rhodamine-123 stain. From the results, we could able to confirm that the investigatory molecule formononetin has anticancer activity and in future, the study will propose to evaluate the formononetin action against genetic changes occurs during lung cancer progression.

Keywords: Caspase 3/7, formononetin, lung cancer, Rhodamine-123

Procedia PDF Downloads 208

Authors: Feng-Sheng Wang, Chao-Ting Cheng

Abstract:

Developing a drug from conception to launch is costly and time-consuming. Computer-aided methods can reduce research costs and accelerate the development process during the early drug discovery and development stages. This study developed a fuzzy multi-objective hierarchical optimization framework for identifying potential anticancer targets in a metabolic model. First, RNA-seq expression data of colorectal cancer samples and their healthy counterparts were used to reconstruct tissue-specific genome-scale metabolic models. The aim of the optimization framework was to identify anticancer targets that lead to cancer cell death and evaluate metabolic flux perturbations in normal cells that have been caused by cancer treatment. Four objectives were established in the optimization framework to evaluate the mortality of cancer cells for treatment and to minimize side effects causing toxicity-induced tumorigenesis on normal cells and smaller metabolic perturbations. Through fuzzy set theory, a multiobjective optimization problem was converted into a trilevel maximizing decision-making (MDM) problem. The applied nested hybrid differential evolution was applied to solve the trilevel MDM problem using two nutrient media to identify anticancer targets in the genome-scale metabolic model of colorectal cancer, respectively. Using Dulbecco’s Modified Eagle Medium (DMEM), the computational results reveal that the identified anticancer targets were mostly involved in cholesterol biosynthesis, pyrimidine and purine metabolisms, glycerophospholipid biosynthetic pathway and sphingolipid pathway. However, using Ham’s medium, the genes involved in cholesterol biosynthesis were unidentifiable. A comparison of the uptake reactions for the DMEM and Ham’s medium revealed that no cholesterol uptake reaction was included in DMEM. Two additional media, i.e., a cholesterol uptake reaction was included in DMEM and excluded in HAM, were respectively used to investigate the relationship of tumor cell growth with nutrient components and anticancer target genes. The genes involved in the cholesterol biosynthesis were also revealed to be determinable if a cholesterol uptake reaction was not induced when the cells were in the culture medium. However, the genes involved in cholesterol biosynthesis became unidentifiable if such a reaction was induced.

Keywords: Cancer metabolism, genome-scale metabolic model, constraint-based model, multilevel optimization, fuzzy optimization, hybrid differential evolution

Procedia PDF Downloads 73

Authors: Abdul Matin, Muhammad Ajmal, Uzma Yunus, Noaman-ul Haq, Hafiz M. Shohaib, Ambreen G. Muazzam

Abstract:

Carbon nanoparticles (CNPs) have unique properties that are useful for the diagnosis and treatment of cancer due to their precise properties like small size (ideal for delivery within the body) stability in solvent and tunable surface chemistry for targeted delivery. Here, highly fluorescent, monodispersed and water-soluble CNPs were synthesized directly from a suitable carbohydrate source (glucose and sucrose) by one-step acid assisted ultrasonic treatment at 35 KHz for 4 hours. This method is green, simple, rapid and economical and can be used for large scale production and applications. The average particle sizes of CNPs are less than 10nm and they emit bright and colorful green-blue fluorescence under the irradiation of UV-light at 365nm. The CNPs were characterized by scanning electron microscopy, fluorescent spectrophotometry, Fourier transform infrared spectrophotometry, ultraviolet-visible spectrophotometry and TGA analysis. Fluorescent CNPs were used as fluorescent probe and nano-carriers for anticancer drug. Functionalized CNPs (with ethylene diamine) were attached with anticancer drug-Methotrexate. In vitro bioactivity and biocompatibility of CNPs-drug conjugates was evaluated by LDH assay and Sulforhodamine B assay using human lung carcinoma cell lines (H157). Our results reveled that CNPs showed biocompatibility and CNPs-anticancer drug conjugates have shown potent cytotoxic effects and high antitumor activities in lung cancer cell lines. CNPs are proved to be excellent substitute for conventional drug delivery cargo systems and anticancer therapeutics in vitro. Our future studies will be more focused on using the same nanoparticles in vivo model system.

Keywords: carbon nanoparticles, carbon nanoparticles-methotrexate conjugates, human lung carcinoma cell lines, lactate dehydrogenase, methotrexate

Procedia PDF Downloads 299

Authors: Ali Azam Talukder, Jamsheda Ferdous Tuli, Tanzina Islam Reba, Shuvra Kanti Dey, Mamoru Yamada

Abstract:

Recent global warming created by various pollutants prompted us to find new energy sources instead of fossil fuels. Fossil fuels are one of the key factors to emit various toxic gases in this planet. To solve this problem, along with the scarcity of the worldwide energy crisis, scientists are looking for various alternative options to mitigate the necessity of required future fuels. In this context, bioethanol can be one of the most suitable alternative energy sources. Bioethanol is a renewable, environment-friendly and carbon-neutral sustainable energy. In our previous study, we identified several bioethanol-producing microbes from the natural fermented sources of Bangladesh. Among them, the strain 4C encoded Saccharomyces cerevisiae produced maximum bioethanol when the fermentation temperature was 25˚C. In this study, we have established high-temperature simultaneous saccharification and fermentation process (HTSSF) by co-culturing of thermally adapted thermosensitive 4C as a fermenting agent and Bacillus amyloliquefaciens (C7), as a saccharifying agent under various physiological conditions or treatments. Conventional methods were applied for cell culture, media preparation and other experimental purposes. High-temperature adaptation of strain 4C was made from 30-42ᵒC, using either YPD or YPS media. In brief, for thermal adaptation, the temperature was periodically increased by 2ᵒC, 1ᵒC and 0.5ᵒC when medium growth temperatures were 30-36ᵒC, 36-40ᵒC, and 40-42ᵒC, respectively, where applicable. Amylase activity and bioethanol content were measured by DNS (3, 5-dinitrosalicylic acid) and solvent extraction and dichromate oxidation method, respectively. Among the various growth parameters like temperatures (30˚C, 37˚C and 42˚C), pHs (5.0, 6.0 and 7.0), carbon sources (5.0-10.0%) and ethanol stress tolerance (0.0-12.0%) etc. were tested, maximum Amylase activity (4.0 IU/ml/min) was recorded for Bacillus amyloliquefaciens (C7) at 42˚C, pH 6.0 and 10% starch. On the other hand, 4.10% bioethanol content was recorded when the thermally adapted strain 4C was co-cultured with C7 at 37ᵒC, pH 6.0 and 10.0% starch for 72 hours at HTSSF process. On the other hand, thermally non-adapted strains gave only 0.5-2.0% bioethanol content under the same physiological conditions. The thermally adapted strain 4C and strain C7, both can tolerate ethanol stress up to 12%. Altogether, a comparative study revealed that our established HTSSF process may be suitable for pilot scale and subsequently at industrial level bioethanol production.

Keywords: bioethanol, co-culture, fermentation, saccharification

Procedia PDF Downloads 82

Authors: Hussen Nasir, Mohd Fitri Mansor, Noor Hidayah Abu

Abstract:

Liberation of the SME market led to a dynamic competitive business environment. SMEs are considered as an engine for economic growth of most nations. SMEs must be willing to adopt the latest technology and incorporate innovation as part of their business strategies. The aim of this study is to examine the innovation practices (innovation strategy and innovation culture) towards sustaining SME’s performance and the moderating effect of intellectual capital on the relationship of innovation practices on SME’s performance. The study will identify the strongest variables that influence the SMEs performance and proposed several hypothesis toward innovation and intellectual capital. Finally, the study will propose a theoretical framework. The current study will contribute to the knowledge of important on the element of intellectual capital as well as innovation practices in the SMEs performance.

Keywords: intellectual capital, innovation practices, SMEs, performance, innovation strategy, innovation culture

Procedia PDF Downloads 467

Authors: Vinaya Kambappa, Chinnadurai Mani, Komaraiah Palle

Abstract:

Non-small cell-lung cancer (NSCLC) cells have increased expression of EGFR, which makes them a potential target for cancer therapy. Based on molecular docking and previous reports, we designed and synthesized quinazoline derivatives as potent EGFR inhibitors. Among the derivatives, three compounds showed good antiproliferative activity against A-549 and H-1299 cells. Furthermore, these compounds inhibited EGFR signaling exhibiting diminishing p-EGFR and its downstream proteins like p-Akt, p-Erk1/2, and p-mTOR; however, it did not alter the levels of EGFR, Akt, Erk1/2 and mTOR proteins. Flow cytometric analysis indicated the accumulation of cells at G1 phase suggesting induction of apoptosis, which was further confirmed by annexin V/propidium iodide staining. Our study suggested that quinazoline scaffold can be developed as novel EGFR kinase inhibitors for cancer therapy.

Keywords: apoptosis, non-small cell-lung cancer cells, EGFR, quinazoline

Procedia PDF Downloads 180

Authors: Arnab Sarkar, Michael Huang, Chuang Ren, Jun Li

Abstract:

The increasing importance of computing in modern society has brought substantial growth in the demand for more computational power. In some problem domains such as scientific simulations, available computational power still sets a limit on what can be practically explored in computation. For many types of code, there is non-uniformity in the utility of computation. That is not every piece of computation contributes equally to the quality of the result. If this non-uniformity is understood well and exploited effectively, we can much more effectively utilize available computing power. In this paper, we discuss a case study of exploring such non-uniformity in a particle-in-cell simulation platform. We find both the existence of significant non-uniformity and that it is generally straightforward to exploit it. We show the potential of order-of-magnitude effective performance gain while keeping the comparable quality of output. We also discuss some challenges in both the practical application of the idea and evaluation of its impact.

Keywords: approximate computing, landau damping, non uniform utility computing, particle-in-cell

Procedia PDF Downloads 255

Authors: Marina Laškarin, Vlado Galičić

Abstract:

Quick adoption of e-business and emerging influence of “Electronic Word of Mouth e-WOM” communication on guests made leading hotel brands successful examples of electronic guest relationship management. Main reasons behind such success are well established procedures in collection, analysis and usage of highly valuable data available on the Internet, generated through some form of e-GRM programme. E-GRM is more than just a technology solution. It’s a system which balance respective guest demands, hotel technological capabilities and organizational culture of employees, discharging the universal approach in guest relations “same for all”. The purpose of this research derives from the necessity of determining the importance of monitoring and applying e-WOM communication as one of the methods used in managing guest relations. This paper analyses and compares different hotelier’s opinions on e-WOM communication.

Keywords: brand loyalty, e-WOM communication, GRM programmes, organizational culture

Procedia PDF Downloads 288

Authors: Jaroslav Kusnir

Abstract:

In her short story collection, Sabrina & Corina, Kali Fajardo-Anstine depicts mostly Latina characters of indigenous background living and travelling in the American West and the Southwest. In all the stories, place and the environment plays an important role in the construction of cultural identity of these characters that is influenced by their indigenous background, a specificity of the American West, its culture and environment, as well as a contemporary (modern) American culture, position of women and gender roles in a Latino community in the USA. This paper will analyze Fajardo-Anstine´s depiction of a specificity of place, especially of the American West and its role in a construction of Latino/a cultural identity in a modern American society as manifested especially in Fajardo-Anstine´s stories Any Further West and Sabrina & Corina. At the same time, the paper will point out Fajardo-Anstine´s construction of cultural identity of female characters and their gender roles in both Latino and a contemporary American societies. The research results show that the formation of Latina cultural identity is closely connected with both place, that is the American West and the Soutwest as well as with Latina and contemporary American cultures.

Keywords: American culture, american west, cultural identity, female identity, latina identity, place

Procedia PDF Downloads 82

Authors: Nguyen Duc An

Abstract:

To the movement towards learning community, creating friendly, positive and appropriate learning environments which best suit the local features is the most salient and decisive factor of the development and success of that learning society. With the aim at building such an English language learning community for the inhabitants in Moc Chau - the national tourist zone, Tay Bac University has successfully designed and deployed the program ‘English in Tourism’ in the project ‘English for Community’. With the strong attachment to the local reality and close knit to the certain communicative situations, this program which was carefully designed and compiled with interesting and practical activities, has greatly helped the locals confidently introduce and popularize the natural beauty, unique culture and specific characteristics of Moc Chau to the foreign tourists; in addition, reinforce awareness of the native culture of the local people as well as improve the professional development in tourism and service.

Keywords: English for community, learning society, learning community, English in tourism

Procedia PDF Downloads 365

Authors: Natalie Riley, Anita Quigley, Robert M. I. Kapsa, George W. Greene

Abstract:

As the fields of medicine and bionics grow rapidly in technological advancement, the future and success of it depends on the ability to effectively interface between the artificial and the biological worlds. The biggest obstacle when it comes to implantable, electronic medical devices, is maintaining a ‘clean’, low noise electrical connection that allows for efficient sharing of electrical information between the artificial and biological systems. Implant fouling occurs with the adhesion and accumulation of proteins and various cell types as a result of the immune response to protect itself from the foreign object, essentially forming an electrical insulation barrier that often leads to implant failure over time. Lubricin (LUB) functions as a major boundary lubricant in articular joints, a unique glycoprotein with impressive anti-adhesive properties that self-assembles to virtually any substrate to form a highly ordered, ‘telechelic’ polymer brush. LUB does not passivate electroactive surfaces which makes it ideal, along with its innate biocompatibility, as a coating for implantable bionic electrodes. It is the aim of the study to investigate LUB’s anti-fouling properties and its potential as a safe, bioinspired material for coating applications to enhance the performance and longevity of implantable medical devices as well as reducing the frequency of implant replacement surgeries. Native, bovine-derived LUB (N-LUB) and recombinant LUB (R-LUB) were applied to gold-coated mylar surfaces. Fibroblast, chondrocyte and neural cell types were cultured and grown on the coatings under both passive and electrically stimulated conditions to test the stability and anti-adhesive property of the LUB coating in the presence of an electric field. Lactate dehydrogenase (LDH) assays were conducted as a directly proportional cell population count on each surface along with immunofluorescent microscopy to visualize cells. One-way analysis of variance (ANOVA) with post-hoc Tukey’s test was used to test for statistical significance. Under both passive and electrically stimulated conditions, LUB significantly reduced cell attachment compared to bare gold. Comparing the two coating types, R-LUB reduced cell attachment significantly compared to its native counterpart. Immunofluorescent micrographs visually confirmed LUB’s antiadhesive property, R-LUB consistently demonstrating significantly less attached cells for both fibroblasts and chondrocytes. Preliminary results investigating neural cells have so far demonstrated that R-LUB has little effect on reducing neural cell attachment; the study is ongoing. Recombinant LUB coatings demonstrated impressive anti-adhesive properties, reducing cell attachment in fibroblasts and chondrocytes. These findings and the availability of recombinant LUB brings into question the results of previous experiments conducted using native-derived LUB, its potential not adequately represented nor realized due to unknown factors and impurities that warrant further study. R-LUB is stable and maintains its anti-fouling property under electrical stimulation, making it suitable for electroactive surfaces.

Keywords: anti-fouling, bioinspired, cell attachment, lubricin

Procedia PDF Downloads 119

Authors: Roxane A. Legaie, Kjiana E. Schwab, Caroline E. Gargett

Abstract:

Studies looking at the changes in gene expression from RNAseq data often make use of linear models. It is also common practice to focus on a subset of data for a comparison of interest, leaving aside the samples not involved in this particular comparison. This work shows the importance of including all observations in the modeling process to better estimate variance parameters, even when the samples included are not directly used in the comparison under test. The human endometrium is a dynamic tissue, which undergoes cycles of growth and regression with each menstrual cycle. The mesenchymal stem cells (MSCs) present in the endometrium are likely responsible for this remarkable regenerative capacity. However recent studies suggest that MSCs also plays a role in the pathogenesis of endometriosis, one of the most common medical conditions affecting the lower abdomen in women in which the endometrial tissue grows outside the womb. In this study we compared gene expression profiles between MSCs and non-stem cell counterparts (‘non-MSC’) obtained from women with (‘E’) or without (‘noE’) endometriosis from RNAseq. Raw read counts were used for differential expression analysis using a linear model with the limma-voom R package, including either all samples in the study or only the samples belonging to the subset of interest (e.g. for the comparison ‘E vs noE in MSC cells’, including only MSC samples from E and noE patients but not the non-MSC ones). Using the full dataset we identified about 100 differentially expressed (DE) genes between E and noE samples in MSC samples (adj.p-val < 0.05 and |logFC|>1) while only 9 DE genes were identified when using only the subset of data (MSC samples only). Important genes known to be involved in endometriosis such as KLF9 and RND3 were missed in the latter case. When looking at the MSC vs non-MSC cells comparison, the linear model including all samples identified 260 genes for noE samples (including the stem cell marker SUSD2) while the subset analysis did not identify any DE genes. When looking at E samples, 12 genes were identified with the first approach and only 1 with the subset approach. Although the stem cell marker RGS5 was found in both cases, the subset test missed important genes involved in stem cell differentiation such as NOTCH3 and other potentially related genes to be used for further investigation and pathway analysis.

Keywords: differential expression, endometriosis, linear model, RNAseq

Procedia PDF Downloads 429