Day 2 :
- Cancer Cell Biology
Introduction: Pathological stage and grade are important parameters for determining the diagnosis and prognosis of urothelial carcinomas. However, this method for interpretation of tumours deemed insufficient due to the heterogenous nature of urothelial carcinomas. Therefore, the utilization of some immunohistochemical markers have been used to improve the criteria. Although some studies have demonstrated an association with certain markers, others using similar markers have been less convincing. In Kuwait, the diagnosis of urothelial carcinoma still depends on the interpretation of hematoxylin and eosin staining. Thus, including reliable immunohistochemical markers will help confirm the diagnosis and aid in the prognosis of the disease. Aim: to correlate the immunohistochemical expression of CK20, Ki67 and mutant p53 in relation to grade and stage of urothelial carcinoma in a randomly selected Kuwaiti population sample. Methodology: A total of 60 paraffin wax embedded samples (TURBT samples) were collected from 60 patients, which consisted of stages; Ta, T1 and T2, were all immunostained by using Ki67, CK20 and mutant p53 antibodies. Results: According to the statistical analysis, there was a statistical significance association with Ki67 positive expression with grade and stage. In addition, a statistical significant association of CK20 diffused expression was observed with grade and stage. A statistical significant association was also observed with positive expression of mtp53 with grade, but no association was found with stage. Conclusion: Overexpression of Ki67 and aberrant expression of CK20 prove to be useful in immunohistochemical tools for predicting prognosis of the disease. Overexpression of mtp53 demonstrated its significance in grade but still remains controversial.
- Cancer Innovations
Université Libre de Bruxelles, Belgium
Mrs Carvalho obtained her Masters degree in Pharmacy in 2012 at the Federal University of Santa Catarina (Brazil). She is now pursuing a PhD at the Université Libre de Bruxelles (Belgium) with a scholarship granted by CAPES/Brazil. Her research interest encompasses the broad field of drug discovery and development. More specifically, her current project aims at deciphering the mechanism of action of a novel beta-carboline derivative as a protein synthesis inhibitor of cancer cells.
Statement of the problem: Gliomas represent the most common primary brain tumors, accounting for 81% of malignant brain tumors . The poor prognosis is partially due to the therapeutic resistance developed by the cells , while the development of new drugs for these tumors faces challenges, such as effective drug delivery (crossing the blood-brain-barrier, BBB) . Evidence indicates that protein synthesis plays an important role in cancer onset and progression . In this project we have investigated the effects of a potential anti-cancer agent, CM16: a new harmine-derivative acting on protein synthesis of Hs683 glioma cells in vitro. Methodology: Two major approaches, molecular biology and proteomics, were used in this work to investigate CM16 effects on the glioma cell line Hs683. Transcription and translation phases were investigated through metabolic labeling assays and further evaluated by sucrose gradients and protein expression level analysis. The study also includes evaluation conducted by the National Cancer Institute (NCI). To study the effects of CM16 on the glioma cells proteome, shotgun proteomics and 2-D electrophoresis were employed. Findings: CM16 is cytostatic at its IC50 concentration and is more selective towards cancerous than non-cancerous cell lines. The growth inhibition profile of the NCI 60-cell-line to CM16 correlates with those of other protein synthesis inhibitors. CM16 induces inhibition of protein synthesis (Figure 1) and these effects likely occur at the initiation phase of translation. Proteome investigation showed significantly different patterns of Hs683 cells treated with CM16 for 15h and 24h (Figure 1). Several proteins involved in the global effects of CM16 on the glioma cells were evidenced with the proteomics evaluation, such as. Conclusion & Significance: CM16 is a potential new anti-cancer agent with the ability to cross the BBB (demonstrated previously in a theoretical model ) acting on the protein synthesis
(initiation phase) of Hs683 cells. Proteomics evaluation revealed that CM16 does not lead to unspecific down regulation of proteins, it rather affects specific proteins.
- Approaches in Cancer Therapy
Aude Ingels is a PhD student in Laboratoire de Cancérologie et de Toxicologie Expérimentale in Université Libre de Bruxelles (Belgium). This Belgian-born biologist started her career in Laboratory of Bone and metabolic biochemistry (ULB) where she studied the physiopathology of nonunion fractures and bone cell therapies. After a short passage as study coordinator (oncology section) in Cliniques Universitaires Saint-Luc (Brussels, Belgium), she started in 2013, a thesis focusing on the use of natural product as anticancer therapy.
To reduce resistance, combinations of therapies with different mechanisms of action/targets are encouraged and studied. Natural products derived from plants have been widely used as source of drugs for diverse medicinal purposes including cancers. We identified sphaeropsidin A as an interesting candidate regarding its specific activity against melanoma. This fungal metabolite revealed to be a modulator of ionic transporters. Thereby it can disrupt the regulatory volume increase process that contributes to protect cells from apoptosis via the regulation of the intracellular chloride concentration. The co-administration of sphaeropsidin A with pro-apoptotic agents could potentially reduce therapies resistance. Aim and methodology: Leaning on this assumption, we evaluated sphaeropsidin A in combination with cytotoxics, e.g. cisplatin or temozolomide with the aim to obtain the maximal cytotoxicity against melanoma cell lines. Owing to the complexity of optimizing the experimental method, we developed a predictive model based on response surface methodology to determine the optimal concentrations of drugs. Findings and conclusions: We found 75 µM of cisplatin and 850 µM of temozolomide combined with 4 to 6 µM of sphaeropsidin A reached maximal in vitro cytotoxic effects. Those concentrations are in line with the in vitro active concentrations of these anti-cancer drugs. Importantly, sphaeropsidin A acted with cisplatin and temozolomide in a synergistic manner. We are now undertaking to compare the cytotoxic activity of sphaeropsidin A with hemisynthetic derivatives with the aim to improve its efficiency in vitro and/ or develop chemical combined drug. Image Figure 1 : Sphaeropsidin A chemical structure. Recent Publications 1. Sullivan RJ, Flaherty KT (2013) Resistance to BRAFtargeted therapy in melanoma. Eur J Cancer 49:1297– 1304. 2. Grazia G, Penna I, Perotti V, et al (2014) Towards combinatorial targeted therapy in melanoma: From preclinical evidence to clinical application (Review). Int J Oncol 45:929–949. 3. Mathieu V, Chantôme A, Lefranc F, et al (2015) Sphaeropsidin A shows promising activity against drugresistant cancer cells by targeting regulatory volume increase. Cell Mol Life Sci 72:3731–3746 4. Dejaegher B, Vander Heyden Y (2011) Experimental designs and their recent advances in set-up, data interpretation, and analytical applications. J Pharm Biomed Anal 56:141–158 5. Ingels A, Dinhof C, Garg A, Maddau L, Masi M, Evidente A, Berger W, Dejaegher B, and Mathieu V (2017) Computed determination of the in vitro optimal chemocombinations of sphaeropsidin A with chemotherapeutic agents to combat melanomas. Cancer Chemother and Pharmacol
- Cancer Screening Tests
Hashemite University, Department of Medical Laboratory Sciences
Breast cancer is the most common cancer among females. Breast cancer is curable during its early stages and is lethal during its late stages. Leptin are hormones produced by adipose tissues with opposite effects on tumor growth, The while leptin promotes tumor development and metastasis.
Aims: This study is aimed to assess the possible association of leptin levels with breast cancer risk, in addition to investigate the relationship between LEP (-2548 G/A) and LEPR (Q223R) polymorphism and breast cancer risk among Jordanian females. Methods: A total of 60 blood samples collected from breast cancer patients were analyzed to identify; the serum level of leptin, leptin and Leptin receptor related polymorphism. A total of 40 healthy females were enrolled as control group. Serum level of leptin were detected by enzyme-linked immunosorbant assay (ELISA), DNA from patients and control were amplified and analyzed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay. Results: The distribution of the two polymorphisms in the studied population were consistent with the Hardy-Weinberg equilibrium. Leptin level was significantly lower in breast cancer patients compared to the controls (p0.05). Conclusion: Our results showed a significant association between low leptin levels in serum and Q223R polymorphism (in leptin receptor gene) with an increase of breast cancer risk. Moreover, No association was found between -2548 G/A polymorphism (in leptin gene) with breast cancer risk. Further studies with larger sample size are suggested.
- Nanotechnology in Cancer
From 1995 to 2000, after 20 years of research using all of my collective information, I did by my own funds in millions /helped only by my family and money I earned from my different small business/ of US $, I was able to realize my goal of creating a device to heal humanity. Using quantum mechanics in physic, electromagnetic science and Chinese Acupuncture Science in Medicine making comparison with west US science in Medicine, I created my life's vision and destiny. I canceled the magnetic aspect of Nikola Tesla AC industrial electricity to create a proprietary invention and discovery of bio-electricity. This bio-electricity has the ability to release bio electron photons which can cleanse plaque in entire vascular system including tiny capillaries’, as well as dead cells (oxidized protein) and other sick cells located in the fiber tissue of any of human physical organ.
The K-BTE device simply releases bio electrons photons. Enriched by natural acids these bio electrons photons become capable of dissolving unhealthy particles in fiber tissue. Then dissolving and dispersing unhealthy particles, it means number of sick particles, dead cells and oxidized proteins from fiber tissue without harm to the healthy cells. This dissolving and dispersing of extreme bad and dangerous unhealthy particles can operate in the brain or any other physical organ. In the brain, any kind of neurological disorders can be checked searching for deep painfully affected areas and then reversed by enriched bio electron photons. The same healing capability occurs identically in an atrophied muscle, bone and cartilage tissue. Also works for plaque in the entire vascular system by dispersing and healing. The ever present keynote of the K-BTE device is always the lack of harm to healthy cells.
The K-BTE device attracts and transfers light of sun quanta particles consisting by its structure of element concentric rings which are consisted by bio-chemical charges of bio photons and electron neutrinos penetrating directly into the brain. This light injection fosters the regeneration, recovery and re-growth of neurons in the brain. This enlightening process facilitates the brain tissue to regain its ability to send impulses throughout the body while simultaneously improving mobility and homeostasis.
- Oncology Nursing
Krishna Institute of Medical Sciences University, Karad, India
Radiotherapy, (RT) a vital modality of treatment in patients with head and neck malignancy (HNM) is associated with high rates of muco-toxicity with no consensus/ universal standard of its care.
Aloe-vera is FDA approved and is therapeutically used. Studies to-date has had different dimensions. In one study, the researcher Puataweepong et al., 2010 used aloe-vera as a mouth-wash where its effect got limited over a few minutes and in the other research, the researcher Su et al., 2004, used it in oral form where it acted as a systemic agent than a local agent. Precisely, the paucity of robust studies on the potential effects of aloe-vera on RIM and its use in the form of an oral gel is a significant omission.
Purpose: The objective of writing this paper is to discuss the effectiveness of oral aloe-vera-gel in reducing the magnitude of increasing RIM during the progress of RTin HNM patients using a double-blind, randomized controlled trial
Research Design/Materials & methods: An evaluative, pre-experimental double-blind, control group design was adopted.100 eligible respondents with HNM scheduled to receive conventional RT were block randomized between Group A and Group B group and PIS implemented. Standardized tools were applied to assess the primary outcome and secondary outcome
Results & discussion/Findings & interpretation: Respondents in experimental and control groups were statistically identical in baseline characteristics. (P>0.05).
The RIM grades were significantly less for experimental group as compared to that for control group (P<0.05).
By the end of treatment, the two groups were statistically different in maximal grade of toxicity (Z = 1.7, P<0.05), use of drugs, QOL scores, percentage of weight loss and nutritional status.
Research implications: Oral aloe-vera gel was beneficial in reducing the magnitude of increasing RIM without side effects
Novelty/Originality: This research is first of its kind to use aloe-vera in gel form as local application.
- Oncologists Meet
Bangabandhu Sheikh Mujib Medical University
Introduction – Carcinoma cervix is the most common Gynaecological cancer in Bangladesh.Seventy percent of cervical cancer cases of Bangladesh come in stage IIB, III & IV diseases which are inoperable. Only 30% cases can be treated by primary surgery. Type II or class II hysterectomy or Wertheim hysterectomy or modified radical hysterectomy is suitable for the stage IA2 disease .Type III or class III or radical hysterectomy is the commonest operation done for stage IIB1 and stage IIA disease. There are four options for the treatment of stage IIB2 cervical cancer.In addition to primary surgery by radical hysterectomy ,stage IIB2 can be treated by neoadjuvant chemotherapy,concurrent chemoradiotherapy or primary radiotherapy. All the surgical procedures should always be associated with bilateral pelvic lymphadenectomy. Study of histopathological report of radical hysterectomy and bilateral pelvic lymphadenectomy (BLPND) specimen can contribute a lot to the decision making for adjuvant therapy of operable cases of carcinoma cervix . Some important informations were gathered by analysis of 100 histopathological report of radical hysterectomy and BLPND specimen of carcinoma cervix
1) To determine the average no of lymphnodes removed during BLPND.
2) To determine the rate of lymph node involvement.
3) To evaluate the cases of lymph node involvement.
Method - Records of histopathological report of radical hysterectomies done at Bongabandhu Sheikh Mujib Medical University were analyzed. Usually bilateral pelvic lymphadenectomy done either introperitoneal or extra peritoneal route. Histopathological report of all the cases are preserved. Data were collected from the histopathological reports, were compiled and statistically analyzed.
Results – Total number of cases studied were 100. The study cases were collected from November 2011 to May 2012. During the period 105 cases had type III radical Histerectomy with bilateral pelvic lymph node dissection. Five cases could not be included in the study due to inadequate information.
All the cases were clinically staged by examination under anesthesia, cystoscopy and intravenous pyelography. Due to lack of logistic support sigmoidoscopy could not be done. But rectal involvement were excluded by per rectal examination. Among the cases 12% were in the age range of 20- 30 years, 10% were between 71-80 years and 60% were between 31-50 years. Regarding histological type 85% were invasive squamous cell carcinoma, 9% were adenocarcinoma, 2% adeno squmous type. Each of the CIS, papillary adenocarcinoma. Adenaid basal CA and Endometroid adeno carcinoma were 1%.
Distribution of grading of the tumours were 45%, 43%, 8% and 3% of grade I, II, III and no grading respectively. Other parameters like number and size of lymph nodes removed, relationship of number and size of lymph nodes with parametrial invasion, involvement of vaginal cuff and lower uterine segment will be presentated at the time of presentation.
Conclusion: Though clinical staging is the norm of management of cervical cancer, the role of histopathological staging is the basis for the adjuvant radiotherapy. In this regard pathologists tremendous contribution cannot be disregarded.
- Recent Diagnostics Technologies
Monterrey Institute of Technology and Higher Education
Perla Pérez-Treviño is a PhD student in Biotechnology from the Tecnologico de Monterrey (ITESM), Mexico. Since 2012 to the present she has been working as a research assistance at the Institute of Cardiology and Vascular Medicine, Zambrano-Hellion Hospital, School of Medicine, ITESM. She has published two papers as first author in peer reviewed journals and two more have been submitted. Perla´s work is focused in the study of molecular and microstructural cell alterations during various chronic pathologies, and currently she is working in assessing the expression of biomarkers in 3D models of cancer cells growth and tumors.
Statement of the Problem: HER2 overexpression is associated with Breast Cancer (BC) poor prognosis, due to increased metastasis and angiogenesis, and decreased apoptosis. HER2 is commonly assessed by immunohistochemistry. Technique that requires extensive sample processing to get thin fixed samples (3-5 mm) that are analyzed using standard HER2 detection probe and subjective algorithms for HER2 interpretation. Consequently, lacks accuracy and reproducibility, and could lead to misdiagnosis. Therefore, we developed a 3D imaging detection method of HER2 using Affibody molecules conjugated with Quantum Dots (Aff-QDs) and ratiometric analysis (RMA) to remove non-specific probe accumulation. Methodology & Theoretical Orientation: Two types of Aff-QDs were prepared; Affibody anti-HER2 and Affibody negative control conjugated by the maleimide reaction with QD605 and QD545, respectively. Both QD were excited at XXX nm, and emitted at the indicated wavelength. Fixed HER2+ and HER2- BC spheroids were incubated with a mixture (1:1) of both Aff-QDs, and confocal image stacks were recorded in the z-axis. Images were processed by RMA (AffantiHER2-QD605/Affneg-QD545 fluorescence), to assess the specific HER2 signal. Findings: We found that the non-specific accumulation for both Aff-QDs was the same within HER2- spheroids. However, the AffantiHER2-QD605 signal in HER2+ spheroids, was significantly higher (5.91 ± 0.81 F/F0) than that of Affneg-QD545 (2.67 ± 0.56 F/F0, p<0.05) and was optimally resolved up to 50 mm depth. After RMA, non-specific signals were removed in HER2+ and HER2- spheroids, and no false HER2 signal was found. Conclusion & Significance: Aff-QDs can efficiently penetrate in spheroids, used as 3D BC models, with minimal sample manipulation; after RMA, specific and objective 3D HER2 result can be obtained. The method proposed here, could reduce the typical problems associated with traditional immunohistochemistry and improve HER2 detection by RMA in thick tissue samples.