Search results for: cutaneous head and neck squamous cell carcinoma

Authors: Mariam Khaled, Noha Farag, Ghada Mohamed Abdel Salam, Khaled Abu-Aisha, Mohamed El-Azizi

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Gastric and colorectal cancers are among the most frequent causes of cancer-associated mortalities in Africa. They have been considered as a global public health concern, as nearly one million new cases are reported per year. IL-1β is a pro-inflammatory cytokine-produced by activated macrophages and monocytes- and a member of the IL-1 family. The inactive IL-1β precursor is cleaved and activated by caspase-1 enzyme, which itself is activated by the assembly of intracellular structures defined as NLRP3 (Nod Like receptor P3) inflammasomes. Activated IL-1β stimulates the Interleukin-1 receptor type-1 (IL-1R1), which is responsible for the initiation of a signal transduction pathway leading to cell proliferation. It has been proven that the IL-1β gene is a highly polymorphic gene in which single nucleotide polymorphisms (SNPs) may affect its expression. It has been previously reported that SNPs including base transitions between C and T at positions, -511 (C-T; dbSNP: rs16944) and -31 (C-T; dbSNP: rs1143627), from the transcriptional start site, contribute to the pathogenesis of gastric and colorectal cancers by affecting IL-1β levels. Altered production of IL-1β due to such polymorphisms is suspected to stimulate an amplified inflammatory response and promote Epithelial Mesenchymal Transition leading to malignancy. Allele frequency distribution of the IL-1β-31 and -511 SNPs, in different populations, and their correlation to the incidence of gastric and colorectal cancers, has been intriguing to researchers worldwide. The current study aims to investigate allele distributions of the IL-1β SNPs among gastric and colorectal cancers Egyptian patients. In order to achieve to that, 89 Biopsy and surgical specimens from the antrum and corpus mucosa of chronic gastritis subjects and gastric and colorectal carcinoma patients was collected for DNA extraction followed by restriction fragment length polymorphism polymerase chain reaction (RFLP-PCR). The amplified PCR products of IL-1β-31C > T and IL-1β-511T > C were digested by incubation with the restriction endonuclease enzymes ALu1 and Ava1. Statistical analysis was carried out to determine the allele frequency distribution in the three studied groups. Also, the effect of the IL-1β -31 and -511 SNPs on nuclear factor binding was analyzed using Fluorescence Electrophoretic Mobility Shift Assay (EMSA), preceded by nuclear factor extraction from gastric and colorectal tissue samples and LPS stimulated monocytes. The results of this study showed that a significantly higher percentage of Egyptian gastric cancer patients have a homozygous CC genotype at the IL-1β-31 position and a heterozygous TC genotype at the IL-1β-511 position. Moreover, a significantly higher percentage of the colorectal cancer patients have a homozygous CC genotype at the IL-1β-31 and -511 positions as compared to the control group. In addition, the EMSA results showed that IL-1β-31C/T and IL-1β-511T/C SNPs do not affect nuclear factor binding. Results of this study suggest that the IL-1β-31 C/T and IL-1β-511 T/C may be correlated to the incidence of gastric cancer in Egyptian patients; however, similar findings couldn’t be proven in the colorectal cancer patients group for the IL-1β-511 T/C SNP. This is the first study to investigate IL-1β -31 and -511 SNPs in the Egyptian population.

Keywords: colorectal cancer, Egyptian patients, gastric cancer, interleukin-1β, single nucleotide polymorphisms

Procedia PDF Downloads 140

Authors: Catherine Felce, Lior Pachter

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Current methods for building phylogenetic trees from gene expression data consider mean expression levels. With single-cell technologies, we can leverage more information about cell dynamics by considering the entire distribution of gene expression across cells. Using biophysical modeling, we propose a method for constructing phylogenetic trees from scRNA-seq data, building on Felsenstein's method of continuous characters. This method can highlight genes whose level of expression may be unchanged between species, but whose rates of transcription/decay may have evolved over time.

Keywords: phylogenetics, single-cell, biophysical modeling, transcription

Procedia PDF Downloads 47

Authors: Venkat Garigapati

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Cell-based therapies are limited due to underlying host immune system activity. Microencapsulation of living cells to overcome this issue has some serious drawbacks, such as limitations of nutrient and oxygen diffusion, which pose a threat to the function and longevity of cells. The conformal coating could overcome the issues which are generally involved in traditional microencapsulation. Some of the theoretical advantages of conformal coating include superior nutrient and oxygen supply to cells, prolonged lifespan, improved drug-secreting cell functionality and an opportunity to load high cell doses in small volumes. Despite several advantages to the conformal coating, there are no suitable methods available to apply to living cells. The ultra-thin conformal coating was achieved utilizing click-reactive methacryloyloxyethyl phosphorylcholine (MPC) polymers, which are capable of specifically reacting one polymer to another at neutral pH in the aqueous isotonic system at the desired temperature suitable for living cells without the need of deleterious initiators. ARPE-19 (Adult Retinal Pigment Epithelial cell line-19) cell-spheroids and rat pancreatic islets were used in the formulation studies. The in vitro studies of coated ARPE-19 cell-spheroids and rat islets indicate that the coat was intact; cells were viable and functioning. The in vitro study results revealed that the conformal coating technology seems promising and in vivo studies are being planned.

Keywords: cells, hydrogel, conformal coating, microencapsulation, insulin

Procedia PDF Downloads 89

Authors: Baljit Singh Khehra, Amar Partap Singh Pharwaha

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Clusters of Microcalcifications (MCCs) are most frequent symptoms of Ductal Carcinoma in Situ (DCIS) recognized by mammography. Least-Square Support Vector Machine (LS-SVM) is a variant of the standard SVM. In the paper, LS-SVM is proposed as a classifier for classifying MCCs as benign or malignant based on relevant extracted features from enhanced mammogram. To establish the credibility of LS-SVM classifier for classifying MCCs, a comparative evaluation of the relative performance of LS-SVM classifier for different kernel functions is made. For comparative evaluation, confusion matrix and ROC analysis are used. Experiments are performed on data extracted from mammogram images of DDSM database. A total of 380 suspicious areas are collected, which contain 235 malignant and 145 benign samples, from mammogram images of DDSM database. A set of 50 features is calculated for each suspicious area. After this, an optimal subset of 23 most suitable features is selected from 50 features by Particle Swarm Optimization (PSO). The results of proposed study are quite promising.

Keywords: clusters of microcalcifications, ductal carcinoma in situ, least-square support vector machine, particle swarm optimization

Procedia PDF Downloads 352

Authors: Leila Ayat, Afak Meftah

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Amorphous-silicon alloys have great promise as low cost solar cell materials. They have excellent photo-conductivity and high optical absorption to sunlight. Now PIN a-Si:H based solar cells are widely used in power generation modules. However, to improve the performance of these cells further, a better fundamental under-standing of the factors limiting cell performance in the homo junction PIN structure is necessary. In this paper we discuss the sensitivity of light J-V characteristics to various device and material parameters in PIN homo junction solar cells. This work is a numerical simulation of the output parameters of a PIN a-Si:H solar cell under AM1.5 spectrum. These parameters are the short circuit current (Jsc), the open circuit voltage (Voc), the fill factor (FF), the conversion efficiency. The simulation was performed with SCAPS-1D software version 3.3 developed at ELIS in Belgium by Marc Burgelman et al. The obtained results are in agreement with experiment. In addition, the effect of the thickness, doping density, capture cross sections of the gap states and the band microscopic mobilities on the output parameters of the cell are also presented.

Keywords: amorphous silicon p-i-n junctions, thin film, solar cells, sensitivity

Procedia PDF Downloads 518

Authors: Rajapaksha Gedara Prasad Tharanga Jayasooriya, Matharage Gayani Dilshara, Yung Hyun Choi, Gi-Young Kim

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Camptothecin (CPT) is a quinolone alkaloid which inhibits DNA topoisomerase I that induces cytotoxicity in a variety of cancer cell lines. We previously showed that CPT effectively inhibited invasion of prostate cancer cells and also combined treatment with subtoxic doses of CPT and TNF-related apoptosis-inducing ligand (TRAIL) potentially enhanced apoptosis in a caspase-dependent manner in hepatoma cancer cells. Here, we found that treatment with CPT caused an irreversible cell cycle arrest in the G2/M phase. CPT-induced cell cycle arrest was associated with a decrease in protein levels of cell division cycle 25C (Cdc25C) and increased the level of cyclin B and p21. The CPT-induced decrease in Cdc25C was blocked in the presence of proteasome inhibitor MG132, thus reversed the cell cycle arrest. In addition to that treatment of CPT-increased phosphorylation of Cdc25C was the resulted of activation of checkpoint kinase 2 (Chk2), which was associated with phosphorylation of ataxia telangiectasia-mutated. Interestingly CPT induced G2/M phase of the cell cycle arrest is reactive oxygen species (ROS) dependent where ROS inhibitors NAC and GSH reversed the CPT-induced cell cycle arrest. These results further confirm by using transient knockdown of nuclear factor-erythroid 2-related factor 2 (Nrf2) since it regulates the production of ROS. Our data reveal that treatment of siNrf2 increased the ROS level as well as further increased the CPT induce G2/M phase cell cycle arrest. Our data also indicate CPT-enhanced cell cycle arrest through the extracellular signal-regulated kinase (ERK) and the c-Jun N-terminal kinase (JNK) pathway. Inhibitors of ERK and JNK more decreased the Cdc25C expression and protein expression of p21 and cyclin B. These findings indicate that Chk2-mediated phosphorylation of Cdc25C plays a major role in G2/M arrest by CPT.

Keywords: camptothecin, cell cycle, checkpoint kinase 2, nuclear factor-erythroid 2-related factor 2, reactive oxygen species

Procedia PDF Downloads 440

Authors: Harry J. Witchel, James Ackah, Carlos P. Santos, Nachiappan Chockalingam, Carina E. I. Westling

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Introduction: Proxemics is the study of how people perceive and use space. It is commonly proposed that when people like or engage with a person/object, they will move slightly closer to it, often quite subtly and subconsciously. Music videos are known to add entertainment value to a pop song. Our hypothesis was that by adding appropriately matched video to a pop song, it would lead to a net approach of the head to the monitor screen compared to simply listening to an audio-only version of the song. Methods: We presented to 27 participants (ages 21.00 ± 2.89, 15 female) seated in front of 47.5 x 27 cm monitor two musical stimuli in a counterbalanced order; all stimuli were based on music videos by the band OK Go: Here It Goes Again (HIGA, boredom ratings (0-100) = 15.00 ± 4.76, mean ± SEM, standard-error-of-the-mean) and Do What You Want (DWYW, boredom ratings = 23.93 ± 5.98), which did not differ in boredom elicited (P = 0.21, rank-sum test). Each participant experienced each song only once, and one song (counterbalanced) as audio-only versus the other song as a music video. The movement was measured by video-tracking using Kinovea 0.8, based on recording from a lateral aspect; before beginning, each participant had a reflective motion tracking marker placed on the outer canthus of the left eye. Analysis of the Kinovea X-Y coordinate output in comma-separated-variables format was performed in Matlab, as were non-parametric statistical tests. Results: We found that the audio-only stimuli (combined for both HIGA and DWYW, mean ± SEM, 35.71 ± 5.36) were significantly more boring than the music video versions (19.46 ± 3.83, P = 0.0066 Wilcoxon Signed Rank Test (WSRT), Cohen's d = 0.658, N = 28). We also found that participants' heads moved around twice as much during the audio-only versions (speed = 0.590 ± 0.095 mm/sec) compared to the video versions (0.301 ± 0.063 mm/sec, P = 0.00077, WSRT). However, the participants' mean head-to-screen distances were not detectably smaller (i.e. head closer to the screen) during the music videos (74.4 ± 1.8 cm) compared to the audio-only stimuli (73.9 ± 1.8 cm, P = 0.37, WSRT). If anything, during the audio-only condition, they were slightly closer. Interestingly, the ranges of the head-to-screen distances were smaller during the music video (8.6 ± 1.4 cm) compared to the audio-only (12.9 ± 1.7 cm, P = 0.0057, WSRT), the standard deviations were also smaller (P = 0.0027, WSRT), and their heads were held 7 mm higher (video 116.1 ± 0.8 vs. audio-only 116.8 ± 0.8 cm above floor, P = 0.049, WSRT). Discussion: As predicted, sitting and listening to experimenter-selected pop music was more boring than when the music was accompanied by a matched, professionally-made video. However, we did not find that the proxemics of the situation led to approaching the screen. Instead, adding video led to efforts to control the head to a more central and upright viewing position and to suppress head fidgeting.

Keywords: boredom, engagement, music videos, posture, proxemics

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

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

Keywords: cell cycle, living cells, nucleus, transcription

Procedia PDF Downloads 310

Authors: Christelle E. Chua, Alicia L. Ho

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The development of a panel of differential gene expression signatures has been of interest in the field of biomarker discovery for radiation exposure. In the absence of the availability of exposed human subjects, lymphocyte cell lines have often been used as a surrogate to human whole blood, when performing ex vivo irradiation studies. The extent of variation between different lymphocyte cell lines is currently unclear, especially with regard to the expression of extracellular miRNA. This study compares the expression profile of extracellular miRNA isolated from different lymphocyte cell lines. It also compares the profile of miRNA obtained when different exosome isolation kits are used. Lymphocyte cell lines were created using lymphocytes isolated from healthy adult males of similar racial descent (Chinese American and Chinese Singaporean) and immortalised with Epstein-Barr virus. The cell lines were cultured in exosome-free cell culture media for 72h and the cell culture supernatant was removed for exosome isolation. Two exosome isolation kits were used. Total exosome isolation reagent (TEIR, ThermoFisher) is a polyethylene glycol (PEG)-based exosome precipitation kit, while ExoSpin (ES, Cell Guidance Systems) is a PEG-based exosome precipitation kit that includes an additional size exclusion chromatography step. miRNA from the isolated exosomes were isolated using miRNEASY minikit (Qiagen) and analysed using nCounter miRNA assay (Nanostring). Principal component analysis (PCA) results suggested that the overall extracellular miRNA expression profile differed between the lymphocyte cell line originating from the Chinese American donor and the cell line originating from the Chinese Singaporean donor. As the gender, age and racial origins of both donors are similar, this may suggest that there are other genetic or epigenetic differences that account for the variation in extracellular miRNA gene expression in lymphocyte cell lines. However, statistical analysis showed that only 3 miRNA genes had a fold difference > 2 at p < 0.05, suggesting that the differences may not be of that great a significance as to impact overall conclusions drawn from different cell lines. Subsequent analysis using cell lines from other donors will give further insight into the reproducibility of results when difference cell lines are used. PCA results also suggested that the method of exosome isolation impacted the expression profile. 107 miRNA had a fold difference > 2 at p < 0.05. This suggests that the inclusion of an additional size exclusion chromatography step altered the subset of the extracellular vesicles that were isolated. In conclusion, these results suggest that extracellular miRNA can be isolated and analysed from exosomes derived from lymphocyte cell lines. However, care must be taken in the choice of cell line and method of exosome isolation used.

Keywords: biomarker, extracellular miRNA, isolation methods, lymphocyte cell line

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Authors: Yessica Nataliani, Miin-Shen Yang

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Cell formation (CF) is an important step in group technology. It is used in designing cellular manufacturing systems using similarities between parts in relation to machines so that it can identify part families and machine groups. There are many CF methods in the literature, but there is less spectral clustering used in CF. In this paper, we propose a spectral clustering algorithm for machine-part CF. Some experimental examples are used to illustrate its efficiency. Overall, the spectral clustering algorithm can be used in CF with a wide variety of machine/part matrices.

Keywords: group technology, cell formation, spectral clustering, grouping efficiency

Procedia PDF Downloads 403

Authors: Hend Mesbah, Yehia Youssef, Ibrahim Hassan, Shaaban Nosier, Ahmed El-Shazly, Ahmed Helal

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The problem of drinking water shortage is becoming more crucial nowadays as a result of the increased demand due to the population growth and the rise in the standard living. In recent years, desalination using electrodialysis powered by solar energy (PV-ED) is being widely used to help provide treated water and reduce the scarcity in water supply. In the present study, a water desalination laboratory scale ED cell with a fixed bed circulation system was designed, developed, and tested. The effect of three parameters (namely, cell voltage , flowrate, and salt concentration) on the removal percentage of salt ions was studied. The cell voltage was adjusted at 3 , 4 and 6 V. A flow rate of 5, 10, and 20 ml/s and an initial salt concentration of 2000, 5000, and 7000 ppm were investigated. The maximum salt percentage removal obtained was 52.5% at the lowest initial concentration (2000 ppm) and at the highest cell voltage (6 V). There was no significant effect of the flow rate on the removal percentage. A model of PV module has also been developed to calculate the dimensions of a solar cell based on the amount of energy consumed and it was calculated from the Overall ED cell voltage.

Keywords: desalination, electrodialysis, solar desalination, photovoltaic electrodialysis

Procedia PDF Downloads 145

Authors: Amin Alvani

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Multiple sclerosis (MS) is a chronic, often disabling disease in which the immune system attacks the myelin sheath of neurons in the central nervous system. The present study aimed to investigate the Relationship between behavioral inhibition/approach system (BIS-BAS) and perceived stress (PS) whit control white blood cell (WBC). 60 MS patients (male=36.7, female=63.3%; age range=15-65 participated in the study and completed the demographic questionnaire, the count blood cell (CBC) test, the behavioral Activation and behavioral inhibition scale (BIS-BAS), and the perceived stress Questionnaire (PSS-14). The results revealed that Between of BAS-reward responsiveness (BAS-DR) subscale and PS, in more than MS patient (BIS), there are increase WBC.

Keywords: behavioral inhibition/approach system, perceived stress, white blood cell, multiple sclerosis

Procedia PDF Downloads 89

Authors: Abhishek Gandhi, Naresh Bhatnagar

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In this article, a new technique has been developed to manufacture open cell extruded thermoplastic foamed sheets with the aid of extrudate surface-quenching phenomenon. As the extrudate foam exits the die, its surface is rapidly quenched which results in freezing of cells on the surface, while the cells at the core continue to grow and leads to development of open-cellular microstructure at the core. Influence of chill roll temperature was found to be extremely significant in developing porous morphological attributes. Subsequently, synergistic effect of blowing agent content and chill roll temperature was examined for their expansion ratio and open-cell microstructure. Further, chill roll rotating speed was found extremely significant in obtaining open-cellular foam structures. This study intends to enhance the understanding of researchers working in the area of open-cell foam processing.

Keywords: foams, porous materials, morphology, composite, microscopy, open-cell foams

Procedia PDF Downloads 447

Authors: Hossein Rassi, Alaheh Gholami Roud-Majany, Zahra Razavi, Massoud Hoshmand

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Cervical cancer is multi step disease that is thought to result from an interaction between genetic background and environmental factors. Human papillomavirus (HPV) infection is the leading risk factor for cervical intraepithelial neoplasia (CIN)and cervical cancer. In other hand, some of hTERT and miRNA polymorphism may plays an important role in carcinogenesis. This study attempts to clarify the relation of hTERT genotypes and miR-146a genotypes in cervical cancer. Forty two archival samples with cervical lesion retired from Khatam hospital and 40 sample from healthy persons used as control group. A simple and rapid method was used to detect the simultaneous amplification of the HPV consensus L1 region and HPV-16,-18, -11, -31, 33 and -35 along with the b-globin gene as an internal control. We use Multiplex PCR for detection of hTERT and miR-146a rs2910164 genotypes in our lab. Finally, data analysis was performed using the 7 version of the Epi Info(TM) 2012 software and test chi-square(x2) for trend. Cervix lesions were collected from 42 patients with Squamous metaplasia, cervical intraepithelial neoplasia, and cervical carcinoma. Successful DNA extraction was assessed by PCR amplification of b-actin gene (99bp). According to the results, hTERT ( rs 2736098) GG genotype and miR-146a rs2910164 CC genotype was significantly associated with increased risk of cervical cancer in the study population. In this study, we detected 13 HPV 18 from 42 cervical cancer. The connection between several SNP polymorphism and human virus papilloma in rare researches were seen. The reason of these differences in researches' findings can result in different kinds of races and geographic situations and also differences in life grooves in every region. The present study provided preliminary evidence that a p53 GG genotype and miR-146a rs2910164 CC genotype may effect cervical cancer risk in the study population, interacting synergistically with HPV 18 genotype. Our results demonstrate that the testing of hTERT rs 2736098 genotypes and miR-146a rs2910164 genotypes in combination with HPV18 can serve as major risk factors in the early identification of cervical cancers. Furthermore, the results indicate the possibility of primary prevention of cervical cancer by vaccination against HPV18 in Iran.

Keywords: polymorphism of hTERT gene, miR-146a rs2910164 polymorphism, cervical cancer, virus

Procedia PDF Downloads 319

Authors: Win Nee Phong, Tau Chuan Ling, Pau Loke Show

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From an industrial perspective, the exploitation of microalgae for protein source is of great economical and commercial interest due to numerous attractive characteristics. Nonetheless, the release of protein from microalgae is limited by the multiple layers of the rigid thick cell wall that generally contain a large proportion of cellulose. Thus an efficient cell disruption process is required to rupture the cell wall. The conventional downstream processing methods which typically involve several unit operational steps such as disruption, isolation, extraction, concentration and purification are energy-intensive and costly. To reduce the overall cost and establish a feasible technology for the success of the large-scale production, microalgal industry today demands a more cost-effective and eco-friendly technique in downstream processing. One of the main challenges to extract the proteins from microalgae is the presence of rigid cell wall. This study aims to provide some guidance on the selection of the efficient solvent to facilitate the proteins released during the cell disruption process. The effects of solvent types such as methanol, ethanol, 1-propanol and water in rupturing the microalgae cell wall were studied. It is interesting to know that water is the most effective solvent to recover proteins from microalgae and the cost is cheapest among all other solvents.

Keywords: green, microalgae, protein, solvents

Procedia PDF Downloads 257

Authors: Chayma Bahar, Chokri Baccouch, Hedi Sakli, Nizar Sakli

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Harvesting and transport of optical and radiofrequency signals are a topical subject with multiple challenges. In this paper, we present a Optical RECTENNA system. We propose here a hybrid system solar cell antenna for 5G mobile communications networks. Thus, we propose rectifying circuit. A parametric study is done to follow the influence of load resistance and input power on Optical RECTENNA system performance. Thus, we propose a solar cell antenna structure in the frequency band of future 5G standard in 2.45 GHz bands.

Keywords: antenna, IoT, optical rectenna, solar cell

Procedia PDF Downloads 177

Authors: Mousa Meratizaman, Sina Monadizadeh, Majid Amidpour

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One of the most favorable thermal desalination methods used widely today is Multi Effect Desalination. High energy consumption in this method causes coupling it with high temperature power cycle like gas turbine. This combination leads to higher energy efficiency. One of the high temperature power systems which have cogeneration opportunities is Solid Oxide Fuel Cell / Gas Turbine. Integration of Multi Effect Desalination with Solid Oxide Fuel Cell /Gas Turbine power cycle in a range of 300-1000 kW is considered in this article. The exhausted heat of Solid Oxide Fuel Cell /Gas Turbine power cycle is used in Heat Recovery Steam Generator to produce needed motive steam for Desalination unit. Thermodynamic simulation and parametric studies of proposed system are carried out to investigate the system performance.

Keywords: solid oxide fuel cell, thermodynamic simulation, multi effect desalination, gas turbine hybrid cycle

Procedia PDF Downloads 376

Authors: Hyun Joon Chang, Jae In Kim, Sungsoo Na

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Kinesin-1 (hereafter called kinesin) is a molecular motor protein that moves cargos toward the end of microtubules using the energy of adenosine triphosphate (ATP) hydrolysis. When kinesin is inactive, its tail autoinhibits the motor chain in order to prevent from reacting with the ATP by cross-linking of the tail domain to the motor domains at two positions. However, the morphological study of kinesin during autoinhibition is yet remained obscured. In this study, we report the effect of the binding site of the tail domain using the normal mode analysis of the elastic network model on kinesin in the tail-free form and tail-bind form. Considering the relationship between the connectivity of conventional network model with respect to the cutoff length and the functionality of the binding site of the tail, we revaluated the network model to observe the key role of the tail domain in its structural aspect. Contingent on the existence of the tail domain, the results suggest the morphological stability of the motor domain. Furthermore, employing the results from normal mode analysis, we have determined the strain energy of the neck linker, an essential portion of the motor domain for ATP hydrolysis. The results of the neck linker also converge to the same indication, i.e. the morphological analysis of the motor domain.

Keywords: elastic network model, Kinesin-1, autoinhibition

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Authors: P. Jost, L. Muckova, M. Matula, J. Pejchal, D. Jun, R. Stetina

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Sulfur mustard (bis(2-chlorethyl) sulfide) is highly toxic, chemical warfare agent that has been used in the past in several armed conflicts. Except for the skin, respiratory tract is one of the important routes of exposure. The elucidation and understanding of the mechanism of toxicity of SM have been effort intensive research. The multiple targets character of SM caused cellular damage resulted in activation of many different mechanisms which contribute to cellular response and participate in the final cytopathology effect. In our present work, we compared time-dependent changes in sulfur mustard exposed adult human lung fibroblasts NHLF and lung epithelial alveolar cell line A-549. Cell viability (MTT assay, Calcein-AM assay, and xCELLigence - real-time cell analysis), apoptosis (flow cytometry), mitochondrial membrane potential (Δψm, flow cytometry), reactive oxygen species induction (DC and cell cycle distribution (flow cytometry) were studied. We observed significantly decreased mitochondrial membrane potential and subsequent induction of apoptosis correlating with decreased cellular viability in the sulfur mustard exposed cells. In low concentrations, sulfur mustard-induced S-phase cell cycle arrest, on the other hand, high concentrations, cell cycle phase distribution of sulfur mustard exposed cells resembled cell cycle phase distribution of control group, which implies nonspecific cell cycle inhibition. Epithelial cells A-549 was found as more sensible to sulfur mustard toxicity. Acknowledgements: This work was supported by a long-term organization development plan Medical Aspects of Weapons of Mass Destruction of the Faculty of Military Health Sciences, University of Defence.

Keywords: apoptosis, cell cycle, cytotoxicity, sulfur mustard

Procedia PDF Downloads 190

Authors: Charlotte K. Lee, Henrique R. N. Aguiar, Ralph Smith, James Baldock, Sam Botchey

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Background: Femoral neck stress fractures (FNSF) are uncommon, making up 1 to 7.2% of stress fractures in healthy subjects. FNSFs are prevalent in young women, military recruits, endurance athletes, and individuals with energy deficiency syndrome or female athlete triad. Presentation is often non-specific and is often misdiagnosed following the initial examination. There is limited research addressing the return–to–activity time after FNSF. Previous studies have demonstrated prognostic time predictions based on various imaging techniques. Here, (1) OxSport clinic FNSF practice standards are retrospectively reviewed, (2) FNSF cohort demographics are examined, (3) Regression models were used to predict return–to–activity prognosis and consequently determine bone stress risk factors. Methods: Patients with a diagnosis of FNSF attending Oxsport clinic between 01/06/2020 and 01/01/2020 were selected from the Rheumatology Assessment Database Innovation in Oxford (RhADiOn) and OxSport Stress Fracture Database (n = 14). (1) Clinical practice was audited against five criteria based on local and National Institute for Health Care Excellence guidance, with a 100% standard. (2) Demographics of the FNSF cohort were examined with Student’s T-Test. (3) Lastly, linear regression and Random Forest regression models were used on this patient cohort to predict return–to–activity time. Consequently, an analysis of feature importance was conducted after fitting each model. Results: OxSport clinical practice met standard (100%) in 3/5 criteria. The criteria not met were patient waiting times and documentation of all bone stress risk factors. Importantly, analysis of patient demographics showed that of the population with complete bone stress risk factor assessments, 53% were positive for modifiable bone stress risk factors. Lastly, linear regression analysis was utilized to identify demographic factors that predicted return–to–activity time [R2 = 79.172%; average error 0.226]. This analysis identified four key variables that predicted return-to-activity time: vitamin D level, total hip DEXA T value, femoral neck DEXA T value, and history of an eating disorder/disordered eating. Furthermore, random forest regression models were employed for this task [R2 = 97.805%; average error 0.024]. Analysis of the importance of each feature again identified a set of 4 variables, 3 of which matched with the linear regression analysis (vitamin D level, total hip DEXA T value, and femoral neck DEXA T value) and the fourth: age. Conclusion: OxSport clinical practice could be improved by more comprehensively evaluating bone stress risk factors. The importance of this evaluation is demonstrated by the population found positive for these risk factors. Using this cohort, potential bone stress risk factors that significantly impacted return-to-activity prognosis were predicted using regression models.

Keywords: eating disorder, bone stress risk factor, femoral neck stress fracture, vitamin D

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Authors: Shiyin He, Zheng Huang

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In this paper, deep learning methods are applied in bio-medical field to detect and count different types of cells in an automatic way instead of manual work in medical practice, specifically in bone marrow examination. The process is mainly composed of two steps, detection and recognition. Mask-Region-Convolutional Neural Networks (Mask-RCNN) was used for detection and image segmentation to extract cells and then Convolutional Neural Networks (CNN), as well as Deep Residual Network (ResNet) was used to classify. Result of cell detection network shows high efficiency to meet application requirements. For the cell recognition network, two networks are compared and the final system is fully applicable.

Keywords: cell detection, cell recognition, deep learning, Mask-RCNN, ResNet

Procedia PDF Downloads 186

Authors: Jaehyung Jung, Kiman Kim, Heesang Eum

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Solar cell is one of the main technologies to reduce greenhouse gas (GHG). Thereby, accurate estimation of greenhouse gas reduction by solar cell technology is crucial to consider strategic applications of the solar cell. The bottom-up approach using operating data such as operation time and efficiency is one of the methodologies to improve the accuracy of the estimation. In this study, alternative GHG reductions from solar cell technology were estimated by a bottom-up approach to indirect emission source (scope 2) in Korea, 2015. In addition, the scenario-based analysis was conducted to assess the effect of technological change with respect to efficiency improvement and rate of operation. In order to estimate GHG reductions from solar cell activities in operating condition levels, methodologies were derived from 2006 IPCC guidelines for national greenhouse gas inventories and guidelines for local government greenhouse inventories published in Korea, 2016. Indirect emission factors for electricity were obtained from Korea Power Exchange (KPX) in 2011. As a result, the annual alternative GHG reductions were estimated as 21,504 tonCO2eq, and the annual average value was 1,536 tonCO2eq per each solar cell technology. Those results of estimation showed to be 91% levels versus design of capacity. Estimation of individual greenhouse gases (GHGs) showed that the largest gas was carbon dioxide (CO2), of which up to 99% of the total individual greenhouse gases. The annual average GHG reductions from solar cell per year and unit installed capacity (MW) were estimated as 556 tonCO2eq/yr•MW. Scenario analysis of efficiency improvement by 5%, 10%, 15% increased as much as approximately 30, 61, 91%, respectively, and rate of operation as 100% increased 4% of the annual GHG reductions.

Keywords: bottom-up approach, greenhouse gas (GHG), reduction, scenario, solar cell

Procedia PDF Downloads 219

Authors: Ganesh K. Maurya, Reema Chaudhary, Neha Pandey, Hari S. Misra

Abstract:

PprA, a pleiotropic protein participating in radioresistance, has been reported for its roles in DNA replication initiation, genome segregation, cell division and DNA repair in polyextremophile Deinococcus radiodurans. Interestingly, expression of deinococcal PprA in E. coli suppresses its growth by reducing the number of colony forming units and provides better resistance against γ-radiation than control. We employed different biochemical and cell biology studies using PprA and its DNA binding/polymerization mutants (K133E & W183R) in E. coli. Cells expressing wild type PprA or its K133E mutant showed reduction in the amount of genomic DNA as well as chromosome copy number in comparison to W183R mutant of PprA and control cells, which suggests the role of PprA protein in regulation of DNA replication initiation in E. coli. Further, E. coli cells expressing PprA or its mutants exhibited different impact on cell morphology than control. Expression of PprA or K133E mutant displayed a significant increase in cell length upto 5 folds while W183R mutant showed cell length similar to uninduced control cells. We checked the interaction of deinococcal PprA and its mutants with E. coli DnaA using Bacterial two-hybrid system and co-immunoprecipitation. We observed a functional interaction of EcDnaA with PprA and K133E mutant but not with W183R mutant of PprA. Further, PprA or K133E mutant has suppressed the ATPase activity of EcDnaA but W183R mutant of PprA failed to do so. These observations suggested that PprA protein regulates DNA replication initiation and cell morphology of surrogate E. coli.

Keywords: DNA replication, radioresistance, protein-protein interaction, cell morphology, ATPase activity

Procedia PDF Downloads 66

Authors: Anna Vildová, H. Hendrychová, J. Kubeš, L. Tůmová

Abstract:

The abiotic elicitation is one of the methods for increasing the secondary metabolites production in plant tissue cultures and it seems to be more effective than traditional strategies. This study verified the use of silver nitrate as elicitor to enhance flavonolignans and flavonoid taxifolin production in suspension culture of Sylibum marianum (L.) Gaertn. Silver nitrate in various concentrations (5.887.10-3 mol/L, 5.887.10-4 mol/L, 5.887.10-5 mol/L) was used as elicitor. The content of secondary metabolites in cell suspension cultures was determined by high performance liquid chromatography. The samples were taken after 6, 12, 24, 48, 72 and 168 hours of treatment. The highest content of taxifolin production (2.2 mg.g-1) in cell suspension culture of Silybum marianum (L.) Gaertn. was detected after silver nitrate (5.887.10-4 mol/L) treatment and 72 h application. Flavonolignans such as silybinA, silybin B, silydianin, silychristin, isosilybin A, isosilybin B were not produced by cell suspension culture of S. marianum after elicitor treatment. Our results show that the secondarymetabolites could be released from S. marianum cells into the nutrient medium by changed permeability of cell wall.

Keywords: Silybum marianum (L.) Gaertn., elicitation, silver nitrate, taxifolin

Procedia PDF Downloads 443

Authors: Sarah Iaquinta, Christophe Blanquart, Elena Ishow, Sylvain Freour, Frederic Jacquemin, Shahram Khazaie

Abstract:

Nanoparticles have recently emerged as a possible cancer treatment tool. Several formulations have been used to enhance the uptake of these nanoparticles by cancer cells and avoid their immediate clearance when administrated in vivo. Most of the previous studies focus on the investigation of the influence of the mechanical properties of the cell membrane and the particle. However, these studies do not account for the variation of adhesion and tension during the wrapping of the nanoparticle by the membrane. These couplings should be considered since the cell adapts to the interaction with the nanoparticle by, e.g., increasing the number of interactions (consequently leading to an increase of the cell membrane/nanoparticle adhesion) and by reorganizing its cytoskeleton, leading to the releasing of the tension of the cell membrane. The main contribution of this work is the proposal of a novel model for representing the cellular uptake of rigid circular nanoparticles based on an energetic model tailored to take into account the adaptation of the nanoparticle/cell membrane adhesion and of the membrane stress during wrapping. Several coupling models using sigmoidal functions are considered and compared. The study calculations revealed that the results considering constant parameters underestimated the final wrapping degree of the particle by up to 50%.

Keywords: adhesion, cellular adaptation, cellular uptake, mechanical properties, tension

Procedia PDF Downloads 210

Authors: Qiang Xu, Xingxin Wu, Wenjie Guo, Xingqi Wang, Yang Sun, Renxiang Tan

Abstract:

The phosphatase SHP-2 is known to exert regulatory activities on cytokine receptor signaling and the dysregulation of SHP-2 has been implicated in the pathogenesis of a variety of diseases. Here we report several small molecule regulators of SHP-2 for the treatment of T cell-mediated diseases. The new cyclodepsipeptide trichomides A, isolated from the fermentation products of Trichothecium roseum, increased the phosphorylation of SHP-2 in activated T cells, and ameliorated contact dermatitis in mice. The trichomides A’s effects were significantly reversed by using the SHP-2-specific inhibitor PHPS1 or T cell-conditional SHP-2 knockout mice. Another compound is a cerebroside Fusaruside isolated from the endophytic fungus Fusarium sp. IFB-121. Fusaruside also triggered the tyrosine phosphorylation of SHP-2, which provided a possible mean of selectively targeting STAT1 for the treatment of Th1 cell-mediated inflammation and led to the discovery of the non-phosphatase-like function of SHP-2. Namely, the Fusaruside-activated pY-SHP-2 selectively sequestrated the cytosolic STAT1 to prevent its recruitment to IFN-R, which contributed to the improvement of experimental colitis in mice. Blocking the pY-SHP-2-STAT1 interaction, with SHP-2 inhibitor NSC-87877 or using T cells from conditional SHP-2 knockout mice, reversed the effects of fusaruside. Furthermore, the fusaruside’s effect is independent of the phosphatase activity of SHP-2, demonstrating a novel role for SHP-2 in regulating STAT1 signaling and Th1-type immune responses.

Keywords: SHP-2, small molecules, T cell, T cell-mediated diseases

Procedia PDF Downloads 311

Authors: Eric Chabrol, Sidonia B.G. Eckle, Renate de Boer, James McCluskey, Jamie Rossjohn, Mirjam H.M. Heemskerk, Stephanie Gras

Abstract:

αβ and γδ T-cells are disparate T-cell lineages that, via their use of either αβ or γδ T-cell antigen receptors (TCRs) respectively, can respond to distinct antigens. Here we characterise a new population of human T-cells, term δβ T-cells, that express TCRs comprising a TCR-δ variable gene fused to a Joining-α/Constant-α domain, paired with an array of TCR-β chains. We characterised the cellular, functional, biophysical and structural characteristic feature of this new T-cells population that reveal some new insight into TCR diversity. We provide molecular bases of how δβ T-cells can recognise viral peptide presented by Human Leukocyte Antigen (HLA) molecule. Our findings highlight how components from αβ and γδTCR gene loci can recombine to confer antigen specificity thus expanding our understanding of T-cell biology and TCR diversity.

Keywords: new delta-beta TCR, HLA, viral peptide, structural immunology

Procedia PDF Downloads 424

Authors: Ana Maria Muț, Georgeta Coneac, Ioana Olariu, Ștefana Avram, Ioana Zinuca Pavel, Ionela Daliana Minda, Lavinia Vlaia, Cristina Adriana Dehelean, Corina Danciu

Abstract:

Oregano essential oil is obtained from different parts of the plant Origanum vulgare (fam. Lamiaceae) and carvacrol and thymol are primary components, widely recognized for their antimicrobial activity, as well as their antiviral and antifungal properties. Poloxamers are triblock copolymers (Pluronic®), formed of three non-ionic blocks with a hydrophobic polyoxypropylene central chain flanked by two polyoxyethylene hydrophilic chains. They are known for their biocompatibility, sensitivity to temperature changes (sol-to-gel transition of aqueous solution with temperature increase), but also for their amphiphilic and surface active nature determining the formation of micelles, useful for solubilization of different hydrophobic compounds such as the terpenes and terpenoids contained in essential oils. Thus, these polymers, listed in European and US Pharmacopoeia and approved by FDA, are widely used as solubilizers and gelling agents for various pharmaceutical preparations, including topical hydrogels. The aim of this study was to investigate the posibility of solubilizing oregano essential oil (OEO) in polymeric micelles using polyoxypropylene (PPO)-polyoxyethylene (PEO)-polyoxypropylene (PPO) triblock polymers to obtain semisolid systems suitable for topical application. A formulation screening was performed, using Pluronic® F-127 in concentration of 20%, Pluronic® L-31, Pluronic® L-61 and Pluronic® L-62 in concentration of 0.5%, 0.8% respectively 1% to obtain the polymeric micelles-based systems. Then, to each selected system, with or without 10% absolute ethanol, 5% or 8% OEO was added. The obtained transparent poloxamer-based hydrogels containing solubilized OEO were further evaluated for pH, rheological characteristics (flow behaviour, viscosity, consistency and spreadability), using consacrated techniques like potentiometric titration, stationary shear flow test, penetrometric method and parallel plate method. Also, in vitro release and permeation of carvacrol from the hydrogels was carried out, using vertical diffusion cells and synthetic hydrophilic membrane and porcine skin respectively. The pH values and rheological features of all tested formulations were in accordance with official requirements for semisolid cutaneous preparations. But, the formulation containing 0.8% Pluronic® L-31, 10% absolute ethanol, 8% OEO and water and the formulation with 1% Pluronic® L-31, 5% OEO and water, produced the highest cumulative amounts of carvacrol released/permeated through the membrane. The present study demonstrated that oregano essential oil can be successfully solubilized in the investigated poloxamer-based hydrogels. These systems can be further investigated as potential topical therapy for cutaneous papillomas. Funding: This research was funded by Project PN-III-P1-1.1-TE2019-0130, Contract number TE47, Romania.

Keywords: oregano essential oil, carvacrol, poloxamer, topical hydrogels

Procedia PDF Downloads 112

Authors: Ganesh K. Maurya, Reema Chaudhary, Neha Pandey, Hari S. Misra

Abstract:

PprA, a pleiotropic protein participating in radioresistance, has been reported for its roles in DNA replication initiation, genome segregation, cell division and DNA repair in polyextremophile Deinococcus radiodurans. Interestingly, expression of deinococcal PprA in E. coli suppresses its growth by reducing the number of colony forming units and provide better resistance against γ-radiation than control. We employed different biochemical and cell biology studies using PprA and its DNA binding/polymerization mutants (K133E & W183R) in E. coli. Cells expressing wild type PprA or its K133E mutant showed reduction in the amount of genomic DNA as well as chromosome copy number in comparison to W183R mutant of PprA and control cells, which suggests the role of PprA protein in regulation of DNA replication initiation in E. coli. Further, E. coli cells expressing PprA or its mutants exhibited different impact on cell morphology than control. Expression of PprA or K133E mutant displayed a significant increase in cell length upto 5 folds while W183R mutant showed cell length similar to uninduced control cells. We checked the interaction of deinococcal PprA and its mutants with E. coli DnaA using Bacterial two-hybrid system and co-immunoprecipitation. We observed a functional interaction of EcDnaA with PprA and K133E mutant but not with W183R mutant of PprA. Further, PprA or K133E mutant has suppressed the ATPase activity of EcDnaA but W183R mutant of PprA failed to do so. These observations suggested that PprA protein regulates DNA replication initiation and cell morphology of surrogate E. coli.

Keywords: DNA replication, radioresistance, protein-protein interaction, cell morphology, ATPase activity

Procedia PDF Downloads 67

Authors: Keisuke Akiyama, Takashi Sakai, Junichiro Koyanagi, Hideki Yoshikawa, Kazuomi Sugamoto

Abstract:

The geometry of acetabular cartilage surface plays an important role in hip joint biomechanics. The aim of this study was to analyze the morphology of acetabular articular cartilage surface in elderly subjects using a 3D-digitizer. Twenty hemipelves from 12 subjects (mean ages 85 years) were scanned with 3D-digitizer. Each acetabular surface model was divided into four regions: anterosuperior (AS), anteroinferior (AI), posterosuperior (PS), and posteroinferior (PI). In the global acetabulum and each region, the acetabular sphere radius and the standard deviation (SD) of the distance from the acetabular sphere center to the acetabular cartilage surface were calculated. In the global acetabulum, the distance between the acetabular surface model and the maximum sphere which did not penetrate over the acetabular surface model was calculated as the inferred femoral head, and then the distribution was mapped at intervals of 0.5 mm. The SD in AS was significantly larger than that in AI (p = 0.006) and PI (p = 0.001). The SD in PS was significantly larger than that in PI (p = 0.005). The closest region (0-0.5 mm) tended to be distributed at anterior or posterosuperior acetabular edge. The contact between the femoral head and acetabulum might start at the periphery of the lunate surface, especially in the anterior or posterosuperior region. From viewpoint of acetabular morphology, the acetabular articular cartilage in the anterior or posterosuperior edge could be more vulnerable due to direct contact mechanism.

Keywords: acetabulum, cartilage, morphology, 3D-digitizer

Procedia PDF Downloads 343