Search results for: liver cancer cells
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 5182

Search results for: liver cancer cells

4252 Induction of Apoptosis by Diosmin through Interleukins/STAT and Mitochondria Mediated Pathway in Hep-2 and KB Cells

Authors: M. Rajasekar, K. Suresh

Abstract:

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

Keywords: diosmin, apoptosis, antioxidant, STAT pathway

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4251 Surface Modified Core–Shell Type Lipid–Polymer Hybrid Nanoparticles of Trans-Resveratrol, an Anticancer Agent, for Long Circulation and Improved Efficacy against MCF-7 Cells

Authors: M. R. Vijayakumar, K. Priyanka, Ramoji Kosuru, Lakshmi, Sanjay Singh

Abstract:

Trans resveratrol (RES) is a non-flavonoid poly-phenolic compound proved for its therapeutic and preventive effect against various types of cancer. However, the practical application of RES in cancer treatment is limited because of its higher dose (up to 7.5 g/day in humans), low biological half life, rapid metabolism and faster elimination in mammals. PEGylated core-shell type lipid polymer hybrid nanoparticles are the novel drug delivery systems for long circulation and improved anti cancer effect of its therapeutic payloads. Therefore, the main objective of this study is to extend the biological half life (long circulation) and improve the therapeutic efficacy of RES through core shell type of nanoparticles. D-α-tocopheryl polyethylene glycol 1000 succinate (vitamin E TPGS), a novel surfactant is applied for the preparation of PEGylated lipid polymer hybrid nanoparticles. The prepared nanoparticles were evaluated by various state of the art techniques such as dynamic light scattering (DLS) technique for particle size and zeta potential, TEM for shape, differential scanning calorimetry (DSC) for interaction analysis and XRD for crystalline changes of drug. Entrapment efficiency and invitro drug release were determined by ultracentrifugation method and dialysis bag method, respectively. Cancer cell viability studies were performed by MTT assay, respectively. Pharmacokinetic studies after i.v administration were performed in sprague dawley rats. The prepared NPs were found to be spherical in shape with smooth surfaces. Particle size and zeta potential of prepared NPs were found to be in the range of 179.2±7.45 to 266.8±9.61 nm and -0.63 to -48.35 mV, respectively. DSC revealed absence of potential interaction. XRD study revealed presence of amorphous form in nanoparticles. Entrapment efficiency was found to be 83.7 % and drug release was found to be in controlled manner. MTT assay showed low MEC and pharmacokinetic studies showed higher AUC of nanoformulaition than its pristine drug. All these studies revealed that the RES loaded PEG modified core-shell type lipid polymer hybrid nanoparticles can be an alternative tool for chemopreventive and therapeutic application of RES in cancer.

Keywords: trans resveratrol, cancer nanotechnology, long circulating nanoparticles, bioavailability enhancement, core shell nanoparticles, lipid polymer hybrid nanoparticles

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4250 Levels of Toxic Metals in Different Tissues of Lethrinus miniatus Fish from Arabian Gulf

Authors: Muhammad Waqar Ashraf, Atiq A. Mian

Abstract:

In the present study, accumulation of eight heavy metals, lead (Pb), cadmium (Cd), manganese (Mn), copper (Cu), zinc (Zn), iron (Fe), nickel (Ni) and chromium (Cr)was determined in kidney, heart, liver and muscle tissues of Lethrinus miniatus fish caught from Arabian Gulf. Metal concentrations in all the samples were measured using Atomic Absorption Spectroscopy. Analytical validation of data was carried out by applying the same digestion procedure to standard reference material (NIST-SRM 1577b bovine liver). Levels of lead (Pb) in the liver tissue (0.60µg/g) exceeded the limit set by European Commission (2005) at 0.30 µg/g. Zinc concentration in all tissue samples were below the maximum permissible limit (50 µg/g) as set by FAO. Maximum mean cadmium concentration was found 0.15 µg/g in the kidney tissues. Highest content of Mn in the studied tissues was seen in the kidney tissue (2.13 µg/g), whereas minimum was found in muscle tissue (0.87 µg/g). The present study led to the conclusion that muscle tissue is the least contaminated tissue in Lethrinus miniatus and consumption of organs should be avoided as much as possible.

Keywords: lethrinus miniatus, arabian gulf, heavy metals, atomic absorption spectroscopy

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4249 Following the Modulation of Transcriptional Activity of Genes by Chromatin Modifications during the Cell Cycle in Living Cells

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

Abstract:

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

Keywords: cell cycle, living cells, nucleus, transcription

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4248 Role of Long Noncoding RNA HULC on Colorectal Carcinoma Progression through Epigenetically Repressing NKD2 Expression

Authors: Shu-Jun Li, Cheng-Cao Sun, De-Jia Li

Abstract:

Recently, long noncoding RNAs (lncRNAs) have been emerged as crucial regulators of human diseases and prognostic markers in numerous of cancers, including colorectal carcinoma (CRC). Here, we identified an oncogenetic lncRNA HULC, which may promote colorectal tumorigenesis. HULC has been found to be up-regulated and acts as oncogene in gastric cancer and hepatocellular carcinoma, but its expression pattern, biological function and underlying mechanism in CRC is still undetermined. Here, we reported that HULC expression is also over-expressed in CRC, and its increased level is associated with poor prognosis and shorter survival. Knockdown of HULC impaired CRC cells proliferation, migration and invasion, facilitated cell apoptosis in vitro, and inhibited tumorigenicity of CRC cells in vivo. Mechanistically, RNA immunoprecipitation (RIP) and RNA pull-down experiment demonstrated that HULC could simultaneously interact with EZH2 to repress underlying targets NKD2 transcription. In addition, rescue experiments determined that HULC oncogenic function is partly dependent on repressing NKD2. Taken together, our findings expound how HULC over-expression endows an oncogenic function in CRC.

Keywords: long noncoding RNA, HULC, NKD2, colorectal carcinoma, proliferation, apoptosis

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4247 Attitudes, Knowledge and Perceptions towards Cervical Cancer Messages among Female University Students

Authors: Anne Nattembo

Abstract:

Cervical cancer remains a major public health problem in developing countries, especially in Africa. Effective cervical cancer prevention communication requires identification of behaviors, attitudes and increasing awareness of a given population; thus this study focused on investigating awareness, attitudes, and behavior among female university students towards cervical cancer messages. The study objectives sought to investigate the communication behavior of young adults towards cervical cancer, to understand female students recognition of cervical cancer as a problem, to identify the frames related to cervical cancer and their impact towards audience communication and participation behaviors, to identify the factors that influence behavioral intentions and level of involvement towards cervical cancer services and to make recommendations on how to improve cervical cancer communication towards female university students. The researcher obtained data using semi-structured interviews and focus group discussions targeting 90 respondents. The semi-structured in-depth interviews were carried out through one-on-one discussions basis using a set of prepared questions among 53 respondents. All interviews were audio-tape recorded. Each interview was directly typed into Microsoft Word. 4 focus group discussions were conducted with a total of 37 respondents; 2 female only groups with 10 respondents in one and 9 respondents in another, 1 mixed with 12 participants 5 of whom were male, and 1 male only group with 6 participants. The key findings show that the participants preferred to receive and access cervical cancer information from doctors although they were mainly receiving information from the radio. In regards to the type of public the respondents represent, majority of the respondents were non-publics in the sense that they did not have knowledge about cervical cancer, had low levels of involvement and had high constraint recognition their cervical cancer knowledge levels. The researcher identified the most salient audience frames among female university students towards cervical cancer and these included; death, loss, and fear. These frames did not necessarily make cervical cancer an issue of concern among the female university students but rather an issue they distanced themselves from as they did not perceive it as a risk. The study also identified the constraints respondents face in responding to cervical cancer campaign calls-to-action which included; stigma, lack of knowledge and access to services as well as lack of recommendation from doctors. In regards to sex differences, females had more knowledge about cervical cancer than the males. In conclusion the study highlights the importance of interpersonal communication in risk or health communication with a focus on health providers proactively sharing cervical cancer prevention information with their patients. Health provider’s involvement in cervical cancer is very important in influencing behavior and compliance of cervical cancer calls-to-action. The study also provides recommendations for designing effective cervical cancer campaigns that will positively impact on the audience such as packaging cervical cancer messages that also target the males as a way of increasing their involvement and more campaigns to increase awareness of cervical cancer as well as designing positive framed messages to counter the negative audience frames towards cervical cancer.

Keywords: cervical cancer communication, health communication, university students, risk communication

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4246 Indenyl and Allyl Palladates: Synthesis, Bonding, and Anticancer Activity

Authors: T. Scattolin, E. Cavarzerani, F. Visentin, F. Rizzolio

Abstract:

Organopalladium compounds have recently attracted attention for their high stability even under physiological conditions and, above all, for their remarkable in vitro cytotoxicity towards cisplatin-resistant cell lines. Among the organopalladium derivatives, those bearing at least one N-heterocyclic carbene ligand (NHC) and the Pd(II)-η³-allyl fragment have exhibited IC₅₀ values in the micro and sub-micromolar range towards several cancer cell lines in vitro and in some cases selectivity towards cancerous vs. non-tumorigenic cells. Herein, a selection of allyl and indenyl palladates were synthesized using a solvent-free method consisting of grinding the corresponding palladium precursors with different saturated and unsaturated azolium salts. All compounds have been fully characterized by NMR, XRD and elemental analyses. The intramolecular H, Cl interaction has been elucidated and quantified using the Voronoi Deformation Density scheme. Most of the complexes showed excellent cytotoxicity towards ovarian cancer cell lines, with I₅₀ values comparable to or even lower than cisplatin. Interestingly, the potent anticancer activity was also confirmed in a high-serous ovarian cancer (HGSOC) patient-derived tumoroid, with a clear superiority of this class of compounds over classical platinum-based agents. Finally, preliminary enzyme inhibition studies of the synthesized palladate complexes against the model TrxR show that the compounds have high activity comparable to or even higher than auranofin and classical Au(I) NHC complexes. Based on such promising data, further in vitro and in vivo experiments and in-depth mechanistic studies are ongoing in our laboratories.

Keywords: anticancer activity, palladium complexes, organoids, indenyl and allyl ligands

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4245 The Effect of Progressive Muscle Relaxation and Sleep Hygiene Education to Change Sleep Quality Index Scores of Patient with Breast Cancer

Authors: Ika Wulansari, Yati Afiyanti, Indang Trihandini

Abstract:

Sleeping disorder experienced by patients with breast cancer can affect the physical, mental, health, and well-being. This study examines the effect of progressive muscle relaxation training and sleep hygiene education to change sleep quality scores of the patient with breast cancer. The study design using quasi-experiment with pre-post test within the control group, involving 62 breast cancer patients using consecutive sampling method in Jakarta. Statistical test results with independent t-test showed a significant difference in score of sleep quality between in intervention group and the control group (6,66±3,815; 9,30±3,334, p-value = 0,005). Progressive muscle relaxation exercise and sleep hygiene education proven to be affective to change the patients sleeping quality, so that it can be an alternative therapeutic option to overcome sleeping disorders.

Keywords: sleeping disorders, breast cancer, progressive muscle relaxation, sleep hygiene education

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4244 Development and Evaluation of a Psychological Adjustment and Adaptation Status Scale for Breast Cancer Survivors

Authors: Jing Chen, Jun-E Liu, Peng Yue

Abstract:

Objective: The objective of this study was to develop a psychological adjustment and adaptation status scale for breast cancer survivors, and to examine the reliability and validity of the scale. Method: 37 breast cancer survivors were recruited in qualitative research; a five-subject theoretical framework and an item pool of 150 items of the scale were derived from the interview data. In order to evaluate and select items and reach a preliminary validity and reliability for the original scale, the suggestions of study group members, experts and breast cancer survivors were taken, and statistical methods were used step by step in a sample of 457 breast cancer survivors. Results: An original 24-item scale was developed. The five dimensions “domestic affections”, “interpersonal relationship”, “attitude of life”, “health awareness”, “self-control/self-efficacy” explained 58.053% of the total variance. The content validity was assessed by experts, the CVI was 0.92. The construct validity was examined in a sample of 264 breast cancer survivors. The fitting indexes of confirmatory factor analysis (CFA) showed good fitting of the five dimensions model. The criterion-related validity of the total scale with PTGI was satisfactory (r=0.564, p<0.001). The internal consistency reliability and test-retest reliability were tested. Cronbach’s alpha value (0.911) showed a good internal consistency reliability, and the intraclass correlation coefficient (ICC=0.925, p<0.001) showed a satisfactory test-retest reliability. Conclusions: The scale was brief and easy to understand, was suitable for breast cancer patients whose physical strength and energy were limited.

Keywords: breast cancer survivors, rehabilitation, psychological adaption and adjustment, development of scale

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4243 Histopathological and Biochemical Investigations of Protective Role of Honey in Rats with Experimental Aflatoxicosis

Authors: Turan Yaman, Zabit Yener, Ismail Celik

Abstract:

The aim of this study was to investigate the antioxidant properties and protective role of honey, considered a part of traditional medicine, against carcinogen chemical aflatoxin (AF) exposure in rats, which were evaluated by histopathological changes in liver and kidney, measuring level of serum marker enzymes [aspartate aminotransferase (AST), alanin aminotransferase (ALT), gamma glutamil transpeptidase (GGT)], antioxidant defense systems [Reduced glutathione (GSH), glutathione reductase (GR), superoxide dismutase (SOD), glutathione-S-transferase (GST) and catalase (CAT)], and lipid peroxidation content in liver, erythrocyte, brain, kidney, heart and lungs. For this purpose, a total of eighteen healthy Sprague-Dawley rats were randomly allocated into three experimental groups: A (Control), B (AF-treated) and C (AF+honey-treated). While rats in group A were fed with a diet without AF, B, and C groups received 25 µg of AF/rat/day, where C group additionally received 1 mL/kg of honey by gavage for 90 days. At the end of the 90-day experimental period, we found that the honey supplementation decreased the lipid peroxidation and the levels of enzyme associated with liver damage, increased enzymatic and non-enzymatic antioxidants in the AF+honey-treated rats. Hepatoprotective and nephroprotective effects of honey is further substantiated by showing almost normal histological architecture in AF+honey-treated group, compared to degenerative changes in the liver and kidney of AF-treated rats. Additionally, honey supplementation ameliorated antioxidant defense systems and lipid peroxidation content in other tissues of AF+honey-treated rats. In conclusion, the present study indicates that honey has a hepatoprotective and nephroprotective effect in rats with experimental aflatoxicosis due to its antioxidant activity.

Keywords: aflatoxicosis, honey, histopathology, malondialdehyde, antioxidant, rat

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4242 Liver Transplantation after Downstaging with Electrochemotherapy of Large Hepatocellular Carcinoma and Portal Vein Tumor Thrombosis: A Case Report

Authors: Luciano Tarantino, Emanuele Balzano, Aurelio Nasto

Abstract:

S.R. 53 years. January 2009: HCV-related cirrhosis, Child-Pugh A5 class, EGDS no aesophageal Varices. No important comorbidities. Treated with PEG-IFN+Ribavirin (march-november 2009) with subsequent sustained virologic response. HCVRNA absent overtime. October 2016 :CT detected small HCC nodule in the VIII segment (diam.=12 mm). Treated with US guided RF-ablation. November 2016 CT: complete necrosis. Unfortunately, the patient dropped out US and CT follow-up controls.September 2018: asthenia and weight loss. CT showed a large tumor infiltrating V-VII-VI segments and complete PVTT of right portal vein and its branches . Surgical Consultation excluded indication to Liver resection and OLT . 23 october 2018: ECT of a large peri-hilar area of the tumor including the PVTT. 1 and 3 months post-treatment CT showed complete necrosis and retraction of the thrombus and residual viable tumor in the peripheral portion of the right lobe . Therefor, the patient was reevaluated for OLT and considered eligible in waiting list . March 2019: CT showed no perihilar or portal vein recurrence and distant progression in the right lobe . March 2019 : Trans-arterial-Radio-therapy (TARE) of the right lobe. Post-treatment CT demonstrated no perihilar or portal vein recurrence and extensive necrosis of the residual tumor . December 2019: CT demonstrated several recurrences of HCC infiltrating the VI and VII segment . Howewer no recurrence was observed at hepatic hilum and in portal vessels . Therefore, on February 2020 the patient received OLT. At 44 months follow-up, no complication or recurrence or liver disfunction have been observed.

Keywords: hepatocellular carcinoma, portal vein tumor thrombosis, interventional ultrasound, liver tumor ablation, liver transplantation

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4241 Levels of Heavy Metals in Different Tissues of Lethrinus Miniatus Fish from Arabian Gulf

Authors: Muhammad Waqar Ashraf

Abstract:

In the present study, accumulation of eight heavy metals, lead (Pb), cadmium (Cd), manganese (Mn), copper (Cu), zinc (Zn), iron (Fe), nickel (Ni) and chromium (Cr)was determined in kidney, heart, liver and muscle tissues of Lethrinus Miniatus fish caught from Arabian Gulf. Metal concentrations in all the samples were measured using Graphite Furnace Atomic Absorption Spectroscopy (GF-AAS). Analytical validation of data was carried out by applying the same digestion procedure to standard reference material (NIST-SRM 1577b bovine liver). Levels of lead (Pb) in the liver tissue (0.60µg/g) exceeded the limit set by European Commission (2005) at 0.30 µg/g. Zinc concentration in all tissue samples were below the maximum permissible limit (50 µg/g) as set by FAO. Maximum mean cadmium concentration was found to be 0.15 µg/g in the kidney tissues. Highest content of Mn in the studied tissues was seen in the kidney tissue (2.13 µg/g), whereas minimum was found in muscle tissue (0.87 µg/g). The present study led to the conclusion that muscle tissue is the least contaminated tissue in Lethrinus Miniatus and consumption of organs should be avoided as much as possible.

Keywords: Arabian gulf, Lethrinus miniatus, heavy metals, atomic absorption spectroscopy

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4240 Sensing of Cancer DNA Using Resonance Frequency

Authors: Sungsoo Na, Chanho Park

Abstract:

Lung cancer is one of the most common severe diseases driving to the death of a human. Lung cancer can be divided into two cases of small-cell lung cancer (SCLC) and non-SCLC (NSCLC), and about 80% of lung cancers belong to the case of NSCLC. From several studies, the correlation between epidermal growth factor receptor (EGFR) and NSCLCs has been investigated. Therefore, EGFR inhibitor drugs such as gefitinib and erlotinib have been used as lung cancer treatments. However, the treatments result showed low response (10~20%) in clinical trials due to EGFR mutations that cause the drug resistance. Patients with resistance to EGFR inhibitor drugs usually are positive to KRAS mutation. Therefore, assessment of EGFR and KRAS mutation is essential for target therapies of NSCLC patient. In order to overcome the limitation of conventional therapies, overall EGFR and KRAS mutations have to be monitored. In this work, the only detection of EGFR will be presented. A variety of techniques has been presented for the detection of EGFR mutations. The standard detection method of EGFR mutation in ctDNA relies on real-time polymerase chain reaction (PCR). Real-time PCR method provides high sensitive detection performance. However, as the amplification step increases cost effect and complexity increase as well. Other types of technology such as BEAMing, next generation sequencing (NGS), an electrochemical sensor and silicon nanowire field-effect transistor have been presented. However, those technologies have limitations of low sensitivity, high cost and complexity of data analyzation. In this report, we propose a label-free and high-sensitive detection method of lung cancer using quartz crystal microbalance based platform. The proposed platform is able to sense lung cancer mutant DNA with a limit of detection of 1nM.

Keywords: cancer DNA, resonance frequency, quartz crystal microbalance, lung cancer

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4239 Breast Cancer Incidence Estimation in Castilla-La Mancha (CLM) from Mortality and Survival Data

Authors: C. Romero, R. Ortega, P. Sánchez-Camacho, P. Aguilar, V. Segur, J. Ruiz, G. Gutiérrez

Abstract:

Introduction: Breast cancer is a leading cause of death in CLM. (2.8% of all deaths in women and 13,8% of deaths from tumors in womens). It is the most tumor incidence in CLM region with 26.1% from all tumours, except nonmelanoma skin (Cancer Incidence in Five Continents, Volume X, IARC). Cancer registries are a good information source to estimate cancer incidence, however the data are usually available with a lag which makes difficult their use for health managers. By contrast, mortality and survival statistics have less delay. In order to serve for resource planning and responding to this problem, a method is presented to estimate the incidence of mortality and survival data. Objectives: To estimate the incidence of breast cancer by age group in CLM in the period 1991-2013. Comparing the data obtained from the model with current incidence data. Sources: Annual number of women by single ages (National Statistics Institute). Annual number of deaths by all causes and breast cancer. (Mortality Registry CLM). The Breast cancer relative survival probability. (EUROCARE, Spanish registries data). Methods: A Weibull Parametric survival model from EUROCARE data is obtained. From the model of survival, the population and population data, Mortality and Incidence Analysis MODel (MIAMOD) regression model is obtained to estimate the incidence of cancer by age (1991-2013). Results: The resulting model is: Ix,t = Logit [const + age1*x + age2*x2 + coh1*(t – x) + coh2*(t-x)2] Where: Ix,t is the incidence at age x in the period (year) t; the value of the parameter estimates is: const (constant term in the model) = -7.03; age1 = 3.31; age2 = -1.10; coh1 = 0.61 and coh2 = -0.12. It is estimated that in 1991 were diagnosed in CLM 662 cases of breast cancer (81.51 per 100,000 women). An estimated 1,152 cases (112.41 per 100,000 women) were diagnosed in 2013, representing an increase of 40.7% in gross incidence rate (1.9% per year). The annual average increases in incidence by age were: 2.07% in women aged 25-44 years, 1.01% (45-54 years), 1.11% (55-64 years) and 1.24% (65-74 years). Cancer registries in Spain that send data to IARC declared 2003-2007 the average annual incidence rate of 98.6 cases per 100,000 women. Our model can obtain an incidence of 100.7 cases per 100,000 women. Conclusions: A sharp and steady increase in the incidence of breast cancer in the period 1991-2013 is observed. The increase was seen in all age groups considered, although it seems more pronounced in young women (25-44 years). With this method you can get a good estimation of the incidence.

Keywords: breast cancer, incidence, cancer registries, castilla-la mancha

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4238 Design and Synthesis of Some Oxadiazole Bearing Benzimidazole Derivatives as Potential Epidermal Growth Factor Receptor Inhibitors

Authors: Ismail Celik, Gulgun Ayhan Kilcigil, Berna Guven, Zumra Kara, Arzu Onay-Besikci

Abstract:

Epidermal Growth Factor Receptor is the cell-surface receptor of the ErbB (erythroblastic leukemia viral oncogene homologue receptors) family of tyrosine kinases. It plays a vital role in regulating the proliferation and differentiation of cells. However, a variety of mechanisms, such as EGFR expression, mutation, and ligand-dependent receptor dimerization, are associated with the development of various activated EGFR tumors. EGFR is highly expressed in most solid tumors, including breast, head and neck cancer, non-small cell lung cancer (NSCLC), renal, ovarian, and colon cancers. Thus, specific EGFR inhibition plays one of the key roles in cancer treatment. The compounds used in the treatment as tyrosine kinase inhibitors are known to contain the benzimidazole isosterium indole, pazopanib, and axitinibin indazole rings. In addition, benzimidazoles have been shown to exhibit protein kinase inhibitory activity in addition to their different biological activities.Based on these data, it was planned and synthesized of some oxadiazole bearing benzimidazole derivatives [N-cyclohexyl-5-((2-phenyl/substitutedphenyl-1H-benzo[d]imidazole-1-yl) methyl)-1,3,4-oxadiazole-2-amine]. EGFR kinase inhibitory efficiency of the synthesized compounds was determined by comparing them with a known kinase inhibitor erlotinib in vitro, and two of the compounds bearing phenyl (19a) and 3,4-dibenzyloxyphenyl (21a) ring exhibited significant activities.

Keywords: benzimidazole, EGFR kinase inhibitory, oxadiazole, synthesis

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4237 Molecular Docking and Synthesis of Nitrogen-Containing Bisphosphonates

Authors: S. Ghalem, M. Mesmoudi, I. Daoudand, H. Allali

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The nitrogen-containing bisphosphonates (N-BPs) are well established as the treatments of choice for disorders of excessive bone resorption, myeloma and bone metastases, and osteoporosis. They inhibit farnesyl pyrophosphate synthase (FFPS), a key enzyme in the mevalonate pathway, resulting in inhibition of the prenylation of small GTP-binding proteins in osteoclasts and disruption of their cytoskeleton, adhesion/spreading, and invasion of cancer cells. A very few examples for synthesis of α-amino bisphosphonates based on several amino acids are known from the literature. In the present work, esters of aminoacid react with ketophsophonate (or their analog acid or acyl) to afford the desired products, α-iminophosphonates. The reaction of imine with dimethyl phosphate in the presence of catalytic amount of I2 give ester of α-aminobisphosphonate as sole product in good yield. Finally, we used computational docking methods to predict how several α-aminobisphosphonates bind to FPPS and how R and X influence. Pamidronate, β-aminobisphosphonate already marketed, was used as reference. These results are of interest since they represent a new and simple way to sythesize α-aminobisphosphonates with a free COOH group increased by R2 functionalisable and opening up the possibility of using the molecular docking to facilitate the design of other, novel FFPS inhibitors.

Keywords: drug research, cancer, α-amino bisphosphonates, molecular docking

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4236 Postmortem Analysis of Lidocaine in Women Died of Criminal Abortion

Authors: Mohammed A. Arishy, Sultan M. Alharbi, Mohammed A. Hakami, Farid M. Abualsail, Mohammad A. Attafi, Riyadh M. Tobaiqi, Hussain M. Alsalem, Ibraheem M. Attafi

Abstract:

Lidocaine is the most common local anesthetics used for para cervical block to reduce pain associated with surgical abortion. A 25-year-old pregnant woman who. She died before reaching hospital, and she was undergoing criminal abortion during the first trimester. In post-mortem investigations and autopsy shows no clear finding; therefore, toxic substances must be suspected and searched for routinely toxicology analysis. In this case report, the postmortem concentration of lidocaine was detected blood, brain, liver, kidney, and stomach. For lidocaine identification and quantification, sample was extracted using solid phase extraction and analyzed by GC-MS (Shimadzu, Japan). Initial screening and confirmatory analysis results showed that only lidocaine was detected in all collected samples, and no other toxic substances or alcohol were detected. The concentrations of lidocaine in samples were 19, 17, 14, 7, and 3 ug/m in the brain, blood, kidney, liver, and stomach, respectively. Lidocaine blood concentration (17 ug/ml) was toxic level and may result in death. Among the tissues, brain showed the highest level of lidocaine, followed by the kidney, liver, and stomach.

Keywords: forensic toxicology, GC-MS, lidocaine, postmortem

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4235 The Role Support Groups Play in Decreasing Depression and PTSD in Cancer Survivors: A Literature Review

Authors: Julianne Macmullen

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Due to advances in technology and early detection and treatment of cancer, many cancer patients are surviving longer than five years post-diagnosis. Most cancer patients suffer from depression, anxiety, and post-traumatic stress disorder (PTSD) at some point during diagnosis, treatment, and survivorship. A subgroup of patients will continue to suffer from depression and PTSD and require early intervention. Support groups provide patients with the emotional and informational support they require while also giving survivors a sense of community, friendship, and purpose. This type of support is recognized by researchers to improve the quality of life while also decreasing depression and PTSD symptoms. The gaps in the literature include cultural diversity, minorities, and support groups involving cancer types other than breast cancer. Another gap in the literature includes the perceptions of cancer patients as well as longitudinal studies to determine the relationships between support groups and decreased depression and PTSD rates over time. Future research is required to fill the gaps in the literature mentioned previously. Future research is also needed to analyze the difference in age groups and different types of support groups such as professionally-led, peer-led, and online. Implications for practice involve providers assessing for the symptoms of depression and PTSD in order to offer prompt treatment and support services to those patients. In conclusion, social support by way of support groups improves the quality of life, gives survivors a sense of purpose to help others while also gaining the support they need, and reduces the rate of depressive episodes related to PTSD.

Keywords: cancer survivor, survivorship, post-traumatic stress disorder (PTSD), depression, support groups

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4234 An Exploration of the Pancreatic Cancer miRNome during the Progression of the Disease

Authors: Barsha Saha, Shouvik Chakravarty, Sukanta Ray, Kshaunish Das, Nidhan K. Biswas, Srikanta Goswami

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Pancreatic Ductal Adenocarcinoma is a well-recognised cause of cancer death with a five-year survival rate of about 9%, and its incidence in India has been found to be increased manifold in recent years. Due to delayed detection, this highly metastatic disease has a poor prognosis. Several molecular alterations happen during the progression of the disease from pre-cancerous conditions, and many such alterations could be investigated for their biomarker potential. MicroRNAs have been shown to be prognostic for PDAC patients in a variety of studies. We hereby used NGS technologies to evaluate the role of small RNA changes during pancreatic cancer development from chronic pancreatitis. Plasma samples were collected from pancreatic cancer patients (n=16), chronic pancreatitis patients (n=8), and also from normal individuals (n=16). Pancreatic tumour tissue (n=5) and adjacent normal tissue samples (n=5) were also collected. Sequencing of small RNAs was carried out after small RNAs were isolated from plasma samples and tissue samples. We find that certain microRNAs are highly deregulated in pancreatic cancer patients in comparison to normal samples. A combinatorial analysis of plasma and tissue microRNAs and subsequent exploration of their targets and altered molecular pathways could not only identify potential biomarkers for disease diagnosis but also help to understand the underlying mechanism.

Keywords: small RNA sequencing, pancreatic cancer, biomarkers, tissue sample

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4233 First Occurrence of Histopathological Assessment in Gadoid Deep-Fish Phycis blennoides from the Southwestern Mediterranean Sea

Authors: Zakia Alioua, Amira Soumia, Zerouali-Khodja Fatiha

Abstract:

In spite of a wide variety of contaminants such as heavy metals and organic compounds in addition to the importance of extended pollution, the deep-sea and its species are not in haven and being affected through contaminants exposure. This investigation is performed in order to provide data on the presence of pathological changes in the liver and gonads of the greater forkbeard. A total of 998 specimens of the teleost fish Phycis blennoides Brünnich, 1768 ranged from 5,7 to 62,7 cm in total length, were obtained from the commercial fisheries of Algerian ports. The sampling has been carried out monthly from December 2013 to June 2015 and from January to June 2016 caught by trawlers and longlines between 75 and 600 fathoms in the coast of Algeria. Individuals were sexed their gonads, and their livers were removed and processed for light microscopy and one case of atresia was identified. In whole, overall 0,002% of the specimens presented some degree of liver steatosis. For the gastric section, 442 selected stomachs contents were observed looking for parasitic infestation and enumerate 212 nematodes. A prospecting survey for metal contaminant was performed on the liver by atomic absorption spectrophotometry analysis.

Keywords: atresia, coast of Algeria, histopathology, nematode, Phycis blennoides, steatosis

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4232 Anticancer Study of Copper and Zinc Complexes with Doxorubicin

Authors: Grzegorz Swiderski, Agata Jablonska-Trypuc, Natalia Popow, Renata Swislocka, Wlodzimierz Lewandowski

Abstract:

Doxorubicin belongs to the group of anthracycline antitumor antibiotics. Because of the wide spectrum of actions, it is one of the most widely used anthracycline antibiotics, including the treatment of breast, ovary, bladder, lung cancers as well as neuroblastoma, lymphoma, leukemia and myeloid leukemia. Antitumor activity of doxorubicin is based on the same mechanisms as for most anthracyclines. Like the metal ions affect the nucleic acids on many biological processes, so the environment of the metal chelates of antibiotics can have a significant effect on the pharmacological properties of drugs. Complexation of anthracyclines with metal ions may contribute to the production of less toxic compounds. In the framework of this study, the composition of complexes obtained in aqueous solutions of doxorubicin with metal ions (Cu2+ and Zn2+). Complexation was analyzed by spectrophotometric titration in aqueous solution at pH 7.0. The pH was adjusted with 0.02M Tris-HCl buffer. The composition of the complexes found was Cu: doxorubicin (1: 2) and a Zn: doxorubicin (1: 1). The effect of Dox, Dox-Cu and Dox-Zn was examined in MCF-7 breast cancer cell line, which were obtained from American Type Culture Collection (ATCC). The compounds were added to the cultured cells for a final concentration in the range of 0,01µM to 0,5µM. The number of MCF-7 cells with division into living and dead, was determined by direct counts of cells with the use of trypan blue dye using LUNA Logos Biosystems cell counter. ApoTox-Glo Triplex Assay (Promega, Madison, Wisconsin, USA) was used according to the manufacturer’s instructions to measure the MCF-7 cells’ viability, cytotoxicity and apoptosis. We observed a decrease in cells proliferation in a dose-dependent manner. An increase in cytotoxicity and decrease in viability in the ApoTox Triplex assay was also showed for all tested compounds. Apoptosis, showed as caspase 3/7 activation, was observed only in Dox treatment. In Dox-Zn and Dox-Cu caspase 3/7 activation was not observed. This work was financially supported by National Science Centre, Poland, under the research project number 2014/13/B/NZ7/02 352.

Keywords: anticancer properties, anthracycline antibiotic, doxorubicine, metal complexes

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4231 Simulation of Remove the Fouling on the in vivo By Using MHD

Authors: Farhad Aalizadeh, Ali Moosavi

Abstract:

When a blood vessel is injured, the cells of your blood bond together to form a blood clot. The blood clot helps you stop bleeding. Blood clots are made of a combination of blood cells, platelets(small sticky cells that speed up the clot-making process), and fibrin (protein that forms a thread-like mesh to trap cells). Doctors call this kind of blood clot a “thrombus.”We study the effects of different parameters on the deposition of Nanoparticles on the surface of a bump in the blood vessels by the magnetic field. The Maxwell and the flow equations are solved for this purpose. It is assumed that the blood is non-Newtonian and the number of particles has been considered enough to rely on the results statistically. Using MHD and its property it is possible to control the flow velocity, remove the fouling on the walls and return the system to its original form.

Keywords: MHD, fouling, in-vivo, blood clots, simulation

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4230 Initiation of Paraptosis-Like PCD Pathway in Hepatocellular Carcinoma Cell Line by Hep88 mAb through the Binding of Mortalin (HSPA9) and Alpha-Enolase

Authors: Panadda Rojpibulstit, Suthathip Kittisenachai, Songchan Puthong, Sirikul Manochantr, Pornpen Gamnarai, Sasichai Kangsadalampai, Sittiruk Roytrakul

Abstract:

Hepatocellular carcinoma (HCC) is the most primary hepatic cancer worldwide. Nowadays a targeted therapy via monoclonal antibodies (mAbs) specific to tumor-associated antigen is continually developed in HCC treatment. In this regard, after establishing and consequently exploring Hep88 mAb’s tumoricidal effect on hepatocellular carcinoma cell line (HepG2 cell line), the Hep88 mAb’s specific Ag from both membrane and cytoplasmic fractions of HepG2 cell line was identified by 2-D gel electrophoresis and western blot analysis. After in-gel digestion and subsequent analysis by liquid chromatography-mass spectrometry (LC-MS), mortalin (HSPA9) and alpha-enolase were identified. The recombinant proteins specific to Hep88 mAb were cloned and expressed in E.coli BL21 (DE3). Moreover, alteration of HepG2 and Chang liver cell line after being induced by Hep88 mAb for 1-3 days was investigated using a transmission electron microscope. The result demonstrated that Hep88 mAb can bind to the recombinant mortalin (HSPA9) andalpha-enolase. In addition, gradual appearance of mitochondria vacuolization and endoplasmic reticulum dilatation were observed. Taken together, paraptosis-like programmed cell death (PCD) of HepG2 is induced by binding of mortalin (HSPA9) and alpha-enolase to Hep88 mAb. Mortalin depletion by formation of Hep88 mAb-mortalin (HSPA9) complex might initiate transcription-independent of p53-mediated apoptosis. Additionally, Hep88 mAb-alpha-enolase complex might initiate HepG2 cells energy exhaustion by glycolysis pathway obstruction. These results imply that Hep88 mAb might be a promising tool for development of an effective treatment of HCC in the next decade.

Keywords: Hepatocellular carcinoma, Monoclonal antibody, Paraptosis-like program cell death, Transmission electron microscopy, mortalin (HSPA9), alpha-enolase

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4229 Biodistribution Studies of 177Lu-DOTATOC in Mouse Tumor Model: Possible Utilization in Adenocarcinoma Breast Cancer Treatment

Authors: M. Mousavi-Daramoroudi, H. Yousefnia, F. Abbasi-Davani, S. Zolghadri, S. Kakaei

Abstract:

Despite the appropriate characteristics of 177Lu and DOTATOC, to our best knowledge, the therapeutic benefit of 177Lu-DOTATOC complex in breast cancer has not been reported until now. In this study, biodistribution of 177Lu-DOTA-TOC in mouse tumor model for evaluation of possible utilization of this complex in breast cancer treatment was investigated.177Lu was prepared with the specific activity of 2.6-3 GBq.mg-1 and radionuclidic purity higher than 99%. The radiolabeled complex was prepared in the optimized conditions with the radiochemical purity higher than 99%. The final solution was injected to the BALB/c mice with adenocarcinoma breast cancer. The biodistribution results showed major accumulation in the kidneys as the major excretion route and the somatostatin receptor-positive tissues such as pancreas compared with the other tissues. Also, significant uptake was observed in tumor even in longer time after injection. According to the results obtained in this research study, somatostatin receptors expressed in breast cancers can be targeted with DOTATOC analogues especially with 177Lu-DOTATOC as an ideal therapeutic agent.

Keywords: ¹⁷⁷Lu, adenocarcinoma breast cancer, DOTATOC, BALB/c mice

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4228 Albumin-Induced Turn-on Fluorescence in Molecular Engineered Fluorescent Probe for Biomedical Application

Authors: Raja Chinnappan, Huda Alanazi, Shanmugam Easwaramoorthi, Tanveer Mir, Balamurugan Kanagasabai, Ahmed Yaqinuddin, Sandhanasamy Devanesan, Mohamad S. AlSalhi

Abstract:

Serum albumin (SA) is a highly rich water-soluble protein in plasma. It is known to maintain the living organisms' health and help to maintain the proper liver function, kidney function, and plasma osmolality in the body. Low levels of serum albumin are an indication of liver failure and chronic hepatitis. Therefore, it is important to have a low-cost, accurate and rapid method. In this study, we designed a fluorescent probe, triphenylamine rhodanine-3-acetic acid (mRA), which triggers the fluorescence signal upon binding with serum albumin (SA). mRA is a bifunctional molecule with twisted intramolecular charge transfer (TICT)-induced emission characteristics. An aqueous solution of mRA has an insignificant fluorescence signal; however, when mRA binds to SA, it undergoes TICT and turns on the fluorescence emission. A SA dose-dependent fluorescence signal was performed, and the limit of detection was found to be less than ng/mL. The specific binding of SA was tested from the cross-reactivity study using similar structural or functional proteins.

Keywords: serum albumin, fluorescent sensing probe, liver diseases, twisted intramolecular charge transfer

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4227 Sitagliptin-AntiCD4 Mab Conjugated T Cell Targeting Therapy for the Effective Treatment of Type I Diabetes

Authors: T. Mahesh, M. K. Samanta

Abstract:

Antibody dug conjugate (ADC’s) concept is a less explored and more trustable for the treatment of Type 1 diabetes (T1D). T1D is thought to arise from selective immunologically mediated destruction of the insulin- producing β-cells in the pancreatic islets of Langerhans with consequent insulin deficiency. It is evident that type 1 diabetes can be conquered, by 1) to stop immune destruction of βcells, 2) to replace or regenerate β-cells, and 3) to preserve β-cell function and mass. Many studies found that the regulatory T cells (Tregs) are crucial for the maintenance of immunological tolerance. Immune tolerance is liable for the activation of the Th1 response. The important role of Th1 response in pathology of T1D entails the depletion of CD4+ T cells, which initiated the use of anti-CD4 monoclonal antibodies (mAbs) against CD4+ T cells to interfere with induction of T1D.Insulin is regulated by Glucagon-Like Peptide-1 hormone (GLP-1) which also stimulates β-cells proliferation as the half-life of GLP-1 harmone is less due to rapid degradation by DPP-IV enzyme an alternative DPP-IV-inhibitors can increase the half-life of GLP-1 through which it conquers the replacement and reserve β-cells mass. Thus in the present study Anti-CD4 mAb was conjugated with Sitagliptin which is a DPP-IV inhibitor Drug loaded in Nanoparticles through Sulfo-MBS cross-linkers. The above study can be an effective approach for treatment to overcome the Passive subcutaneous insulin therapy.

Keywords: antibody drug conjugates, anti-CD4 Mab, DPP IV inhibitors, GLP-1

Procedia PDF Downloads 387
4226 A Single Cell Omics Experiments as Tool for Benchmarking Bioinformatics Oncology Data Analysis Tools

Authors: Maddalena Arigoni, Maria Luisa Ratto, Raffaele A. Calogero, Luca Alessandri

Abstract:

The presence of tumor heterogeneity, where distinct cancer cells exhibit diverse morphological and phenotypic profiles, including gene expression, metabolism, and proliferation, poses challenges for molecular prognostic markers and patient classification for targeted therapies. Understanding the causes and progression of cancer requires research efforts aimed at characterizing heterogeneity, which can be facilitated by evolving single-cell sequencing technologies. However, analyzing single-cell data necessitates computational methods that often lack objective validation. Therefore, the establishment of benchmarking datasets is necessary to provide a controlled environment for validating bioinformatics tools in the field of single-cell oncology. Benchmarking bioinformatics tools for single-cell experiments can be costly due to the high expense involved. Therefore, datasets used for benchmarking are typically sourced from publicly available experiments, which often lack a comprehensive cell annotation. This limitation can affect the accuracy and effectiveness of such experiments as benchmarking tools. To address this issue, we introduce omics benchmark experiments designed to evaluate bioinformatics tools to depict the heterogeneity in single-cell tumor experiments. We conducted single-cell RNA sequencing on six lung cancer tumor cell lines that display resistant clones upon treatment of EGFR mutated tumors and are characterized by driver genes, namely ROS1, ALK, HER2, MET, KRAS, and BRAF. These driver genes are associated with downstream networks controlled by EGFR mutations, such as JAK-STAT, PI3K-AKT-mTOR, and MEK-ERK. The experiment also featured an EGFR-mutated cell line. Using 10XGenomics platform with cellplex technology, we analyzed the seven cell lines together with a pseudo-immunological microenvironment consisting of PBMC cells labeled with the Biolegend TotalSeq™-B Human Universal Cocktail (CITEseq). This technology allowed for independent labeling of each cell line and single-cell analysis of the pooled seven cell lines and the pseudo-microenvironment. The data generated from the aforementioned experiments are available as part of an online tool, which allows users to define cell heterogeneity and generates count tables as an output. The tool provides the cell line derivation for each cell and cell annotations for the pseudo-microenvironment based on CITEseq data by an experienced immunologist. Additionally, we created a range of pseudo-tumor tissues using different ratios of the aforementioned cells embedded in matrigel. These tissues were analyzed using 10XGenomics (FFPE samples) and Curio Bioscience (fresh frozen samples) platforms for spatial transcriptomics, further expanding the scope of our benchmark experiments. The benchmark experiments we conducted provide a unique opportunity to evaluate the performance of bioinformatics tools for detecting and characterizing tumor heterogeneity at the single-cell level. Overall, our experiments provide a controlled and standardized environment for assessing the accuracy and robustness of bioinformatics tools for studying tumor heterogeneity at the single-cell level, which can ultimately lead to more precise and effective cancer diagnosis and treatment.

Keywords: single cell omics, benchmark, spatial transcriptomics, CITEseq

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4225 Unsupervised Segmentation Technique for Acute Leukemia Cells Using Clustering Algorithms

Authors: N. H. Harun, A. S. Abdul Nasir, M. Y. Mashor, R. Hassan

Abstract:

Leukaemia is a blood cancer disease that contributes to the increment of mortality rate in Malaysia each year. There are two main categories for leukaemia, which are acute and chronic leukaemia. The production and development of acute leukaemia cells occurs rapidly and uncontrollable. Therefore, if the identification of acute leukaemia cells could be done fast and effectively, proper treatment and medicine could be delivered. Due to the requirement of prompt and accurate diagnosis of leukaemia, the current study has proposed unsupervised pixel segmentation based on clustering algorithm in order to obtain a fully segmented abnormal white blood cell (blast) in acute leukaemia image. In order to obtain the segmented blast, the current study proposed three clustering algorithms which are k-means, fuzzy c-means and moving k-means algorithms have been applied on the saturation component image. Then, median filter and seeded region growing area extraction algorithms have been applied, to smooth the region of segmented blast and to remove the large unwanted regions from the image, respectively. Comparisons among the three clustering algorithms are made in order to measure the performance of each clustering algorithm on segmenting the blast area. Based on the good sensitivity value that has been obtained, the results indicate that moving k-means clustering algorithm has successfully produced the fully segmented blast region in acute leukaemia image. Hence, indicating that the resultant images could be helpful to haematologists for further analysis of acute leukaemia.

Keywords: acute leukaemia images, clustering algorithms, image segmentation, moving k-means

Procedia PDF Downloads 290
4224 Convolutional Neural Networks versus Radiomic Analysis for Classification of Breast Mammogram

Authors: Mehwish Asghar

Abstract:

Breast Cancer (BC) is a common type of cancer among women. Its screening is usually performed using different imaging modalities such as magnetic resonance imaging, mammogram, X-ray, CT, etc. Among these modalities’ mammogram is considered a powerful tool for diagnosis and screening of breast cancer. Sophisticated machine learning approaches have shown promising results in complementing human diagnosis. Generally, machine learning methods can be divided into two major classes: one is Radiomics analysis (RA), where image features are extracted manually; and the other one is the concept of convolutional neural networks (CNN), in which the computer learns to recognize image features on its own. This research aims to improve the incidence of early detection, thus reducing the mortality rate caused by breast cancer through the latest advancements in computer science, in general, and machine learning, in particular. It has also been aimed to ease the burden of doctors by improving and automating the process of breast cancer detection. This research is related to a relative analysis of different techniques for the implementation of different models for detecting and classifying breast cancer. The main goal of this research is to provide a detailed view of results and performances between different techniques. The purpose of this paper is to explore the potential of a convolutional neural network (CNN) w.r.t feature extractor and as a classifier. Also, in this research, it has been aimed to add the module of Radiomics for comparison of its results with deep learning techniques.

Keywords: breast cancer (BC), machine learning (ML), convolutional neural network (CNN), radionics, magnetic resonance imaging, artificial intelligence

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4223 The Regulation of the Cancer Epigenetic Landscape Lies in the Realm of the Long Non-coding RNAs

Authors: Ricardo Alberto Chiong Zevallos, Eduardo Moraes Rego Reis

Abstract:

Pancreatic adenocarcinoma (PDAC) patients have a less than 10% 5-year survival rate. PDAC has no defined diagnostic and prognostic biomarkers. Gemcitabine is the first-line drug in PDAC and several other cancers. Long non-coding RNAs (lncRNAs) contribute to the tumorigenesis and are potential biomarkers for PDAC. Although lncRNAs aren’t translated into proteins, they have important functions. LncRNAs can decoy or recruit proteins from the epigenetic machinery, act as microRNA sponges, participate in protein translocation through different cellular compartments, and even promote chemoresistance. The chromatin remodeling enzyme EZH2 is a histone methyltransferase that catalyzes the methylation of histone 3 at lysine 27, silencing local expression. EZH2 is ambivalent, it can also activate gene expression independently of its histone methyltransferase activity. EZH2 is overexpressed in several cancers and interacts with lncRNAs, being recruited to a specific locus. EZH2 can be recruited to activate an oncogene or silence a tumor suppressor. The lncRNAs misregulation in cancer can result in the differential recruitment of EZH2 and in a distinct epigenetic landscape, promoting chemoresistance. The relevance of the EZH2-lncRNAs interaction to chemoresistant PDAC was assessed by Real Time quantitative PCR (RT-qPCR) and RNA Immunoprecipitation (RIP) experiments with naïve and gemcitabine-resistant PDAC cells. The expression of several lncRNAs and EZH2 gene targets was evaluated contrasting naïve and resistant cells. Selection of candidate genes was made by bioinformatic analysis and literature curation. Indeed, the resistant cell line showed higher expression of chemoresistant-associated lncRNAs and protein coding genes. RIP detected lncRNAs interacting with EZH2 with varying intensity levels in the cell lines. During RIP, the nuclear fraction of the cells was incubated with an antibody for EZH2 and with magnetic beads. The RNA precipitated with the beads-antibody-EZH2 complex was isolated and reverse transcribed. The presence of candidate lncRNAs was detected by RT-qPCR, and the enrichment was calculated relative to INPUT (total lysate control sample collected before RIP). The enrichment levels varied across the several lncRNAs and cell lines. The EZH2-lncRNA interaction might be responsible for the regulation of chemoresistance-associated genes in multiple cancers. The relevance of the lncRNA-EZH2 interaction to PDAC was assessed by siRNA knockdown of a lncRNA, followed by the analysis of the EZH2 target expression by RT-qPCR. The chromatin immunoprecipitation (ChIP) of EZH2 and H3K27me3 followed by RT-qPCR with primers for EZH2 targets also assess the specificity of the EZH2 recruitment by the lncRNA. This is the first report of the interaction of EZH2 and lncRNAs HOTTIP and PVT1 in chemoresistant PDAC. HOTTIP and PVT1 were described as promoting chemoresistance in several cancers, but the role of EZH2 is not clarified. For the first time, the lncRNA LINC01133 was detected in a chemoresistant cancer. The interaction of EZH2 with LINC02577, LINC00920, LINC00941, and LINC01559 have never been reported in any context. The novel lncRNAs-EZH2 interactions regulate chemoresistant-associated genes in PDAC and might be relevant to other cancers. Therapies targeting EZH2 alone weren’t successful, and a combinatorial approach also targeting the lncRNAs interacting with it might be key to overcome chemoresistance in several cancers.

Keywords: epigenetics, chemoresistance, long non-coding RNAs, pancreatic cancer, histone modification

Procedia PDF Downloads 95