Search results for: transperineal template biopsies of prostate
Commenced in January 2007
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Edition: International
Paper Count: 464

Search results for: transperineal template biopsies of prostate

404 Role of Direct Immunofluorescence in Diagnosing Vesiculobullous Lesions

Authors: Mitakshara Sharma, Sonal Sharma

Abstract:

Vesiculobullous diseases are heterogeneous group of dermatological disorders with protean manifestations. The most important technique for the patients with vesiculobullous diseases is conventional histopathology and confirmatory tests like direct immunofluorescence (DIF) and indirect immunofluorescence (IIF). DIF has been used for decades to investigate pathophysiology and in the diagnosis. It detects molecules such as immunoglobulins and complement components. It is done on the perilesional skin. Diagnosis of DIF test depends on features like primary site of the immune deposits, class of immunoglobulin, number of immune deposits and deposition at other sites. The aim of the study is to correlate DIF with clinical and histopathological findings and to analyze the utility of DIF in the diagnosis of these disorders. It is a retrospective descriptive study conducted for 2 years from 2015 to 2017 in Department of Pathology, GTB Hospital on perilesional punch biopsies of vesiculobullous lesions. Biopsies were sent in Michael’s medium. The specimens were washed, frozen and incubated with fluorescein isothiocyanate (FITC) tagged antihuman antibodies IgA, IgG, IgM, C3 & F and were viewed under fluorescent microscope. Out of 401 skin biopsies submitted for DIF, 285 were vesiculobullous diseases, in which the most common was Pemphigus vulgaris (34%) followed by Bullous pemphigoid (21.5%), Dermatitis herpetiformis (16%), Pemphigus foliaceus (11.9%), Linear IgA disease (11.9%), Epidermolysisbullosa (2.39%) and Pemphigus herpetiformis (1.7%). We will be presenting the DIF findings in the all these vesiculobullous diseases. DIF in conjugation with histopathology gives the best diagnostic yield in these lesions. It also helps in the diagnosis whenever there is a clinical and histopathological overlap.

Keywords: antibodies, direct immunofluorescence, pemphigus, vesiculobullous

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403 Hit-Or-Miss Transform as a Tool for Similar Shape Detection

Authors: Osama Mohamed Elrajubi, Idris El-Feghi, Mohamed Abu Baker Saghayer

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This paper describes an identification of specific shapes within binary images using the morphological Hit-or-Miss Transform (HMT). Hit-or-Miss transform is a general binary morphological operation that can be used in searching of particular patterns of foreground and background pixels in an image. It is actually a basic operation of binary morphology since almost all other binary morphological operators are derived from it. The input of this method is a binary image and a structuring element (a template which will be searched in a binary image) while the output is another binary image. In this paper a modification of Hit-or-Miss transform has been proposed. The accuracy of algorithm is adjusted according to the similarity of the template and the sought template. The implementation of this method has been done by C language. The algorithm has been tested on several images and the results have shown that this new method can be used for similar shape detection.

Keywords: hit-or-miss operator transform, HMT, binary morphological operation, shape detection, binary images processing

Procedia PDF Downloads 298
402 Study on Co-Relation of Prostate Specific Antigen with Metastatic Bone Disease in Prostate Cancer on Skeletal Scintigraphy

Authors: Muhammad Waleed Asfandyar, Akhtar Ahmed, Syed Adib-ul-Hasan Rizvi

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Objective: To evaluate the ability of serum concentration of prostate specific antigen between two cutting points considering it as a predictor of skeletal metastasis on bone scintigraphy in men with prostate cancer. Settings: This study was carried out in department of Nuclear Medicine at Sindh Institute of Urology and Transplantation (SIUT) Karachi, Pakistan. Materials and Method: From August 2013 to November 2013, forty two (42) consecutive patients with prostate cancer who underwent technetium-99m methylene diphosphonate (Tc-99mMDP) whole body bone scintigraphy were prospectively analyzed. The information was collected from the scintigraphic database at a Nuclear medicine department Sindh institute of urology and transplantation Karachi Pakistan. Patients who did not have a serum PSA concentration available within 1 month before or after the time of performing the Tc-99m MDP whole body bone scintigraphy were excluded from this study. A whole body bone scintigraphy scan (from the toes to top of the head) was performed using a whole-body Moving gamma camera technique (anterior and posterior) 2–4 hours after intravenous injection of 20 mCi of Tc-99m MDP. In addition, all patients necessarily have a pathological report available. Bony metastases were determined from the bone scan studies and no further correlation with histopathology or other imaging modalities were performed. To preserve patient confidentiality, direct patient identifiers were not collected. In all the patients, Prostate specific antigen values and skeletal scintigraphy were evaluated. Results: The mean age, mean PSA, and incidence of bone metastasis on bone scintigraphy were 68.35 years, 370.51 ng/mL and 19/42 (45.23%) respectively. According to PSA levels, patients were divided into 5 groups < 10ng/mL (10/42), 10-20 ng/mL (5/42), 20-50 ng/mL (2/42), 50-100 (3/42), 100- 500ng/mL (3/42) and more than 500ng/mL (0/42) presenting negative bone scan. The incidence of positive bone scan (%) for bone metastasis for each group were O1 patient (5.26%), 0%, 03 patients (15.78%), 01 patient (5.26%), 04 patients (21.05%), and 10 patients (52.63%) respectively. From the 42 patients 19 (45.23%) presented positive scintigraphic examination for the presence of bone metastasis. 1 patient presented bone metastasis on bone scintigraphy having PSA level less than 10ng/mL, and in only 1 patient (5.26%) with bone metastasis PSA concentration was less than 20 ng/mL. therefore, when the cutting point adopted for PSA serum concentration was 10ng/mL, a negative predictive value for bone metastasis was 95% with sensitivity rates 94.74% and the positive predictive value and specificities of the method were 56.53% and 43.48% respectively. When the cutting point of PSA serum concentration was 20ng/mL the observed results for Positive predictive value and specificity were (78.27% and 65.22% respectively) whereas negative predictive value and sensitivity stood (100% and 95%) respectively. Conclusion: Results of our study allow us to conclude that serum PSA concentration of higher than 20ng/mL was the most accurate cutting point than a serum concentration of PSA higher than 10ng/mL to predict metastasis in radionuclide bone scintigraphy. In this way, unnecessary cost can be avoided, since a considerable part of prostate adenocarcinomas present low serum PSA levels less than 20 ng/mL and for these cases radionuclide bone scintigraphy could be unnecessary.

Keywords: bone scan, cut off value, prostate specific antigen value, scintigraphy

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401 The Usefulness and Limitations of Manual Aspiration Immediately after Pneumothorax Complicating Percutaneous CT Guided Lung Biopsies: A Retrospective 9-Year Review from a Large Tertiary Centre

Authors: Niall Fennessy, Charlotte Yin, Vineet Gorolay, Michael Chan, Ilias Drivas

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Background: The aim of this study was to evaluate the effect of manual aspiration of air from the pleural cavity in mitigating the need for chest drain placement after a CT-guided lung biopsy. Method: This is a single institution retrospective review of CT-guided lung biopsies performed on 799 patients between September 2013 and May 2021 in a major tertiary hospital. Percutaneous manual aspiration of air was performed in 104/306 patients (34%) with pneumothoraxes as a preventative measure. Simple and multivariate analysis was performed to identify independent risk factors (modifiable and nonmodifiable) for the success of manual aspiration in mitigating the need for chest drain insertion. Results: The overall incidence of pneumothorax was 37% (295/799). Chest drains were inserted for 81/295 (27%) of the pneumothoraxes, representing 81/799 (10%) of all CT-guided lung biopsies. Of patients with pneumothoraces, 104 (36%) underwent percutaneous aspiration via either the coaxial guide needle or an 18 or 20G intravenous catheter attached to a three-way stopcock and syringe. Amongst this group, 13 patients (13%) subsequently required chest drain insertion. The success of percutaneous aspiration in avoiding subsequent pleural drain insertion decreased with aspiration volume >500mL, radial pneumothorax depth >3cm, increased subpleural depth of the lesion, and the presence of background emphysema.

Keywords: computed tomography, lung biopsy, pneumothorax, manual aspiration, chest drainage

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400 Comparative Study of Dermal Regeneration Template Made by Bovine Collagen with and without Silicone Layer in the Treatment of Post-Burn Contracture

Authors: Elia Caldini, Cláudia N. Battlehner, Marcelo A. Ferreira, Rolf Gemperli, Nivaldo Alonso, Luiz P. Vana

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The advent of dermal regenerate templates has fostered major advances in the treatment of acute burns and their sequelae, in the last two decades. Both data on morphological aspects of the newly-formed tissue, and clinical trials comparing different templates, are still lacking. The goal of this study was to prospectively analyze the outcome of patients treated with two of the existing templates, followed by thin skin autograft. They are both made of bovine collagen, one includes a superficial silicone layer. Surgery was performed on patients with impaired mobility resulting from burn sequelae (n = 12 per template). Negative pressure therapy was applied post-surgically; patients were monitored for 12 months. Data on scar skin quality (Vancouver and POSAS evaluation scales), rate of joint mobility recovery, and graft contraction were recorded. Improvement in mobility and skin quality were demonstrated along with graft contraction, in all patients. The silicone-coupled template showed the best performance in all aspects.

Keywords: dermal regeneration template, artificial skin, skin quality, scar contracture

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399 Fabrication and Characterization of Cadmium Sulfide Nanowires on Aluminum Oxide Template

Authors: Malik Imran Afzal

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Cadmium supplied nanowires have unique electrical and optical properties and applications. To obtain cadmium supplied nanowires with regular and good aspect ratio, they can be synthesized by template synthesis method. Porous anodized aluminum oxide is the most promising template with regular hexagonal shapes. Their aspect ratio can be controlled by controlling the pores’ depth and diameter which greatly depend on anodization voltage and temperature of the electrolyte. In this research, high purity aluminium was used to prepare nanotemplates at 5-6°C in 1M phosphoric acid and cadmium supplied was deposited electrochemically using a co-solution of thiourea, cadmium acetate and ammonium acetate. pH was maintained at 11 in a heat bath at 75°C with the help of aqueous ammonia solution. Both porous anodized alumina and cadmium supplied nanowires were characterized suing SEM. A good quality Nanowires were obtained in bunches with reasonably high aspect ratio.

Keywords: bunches, electrodeposition, hexagonal, thiourea

Procedia PDF Downloads 299
398 Oral Antibiotics in Trans-Rectal Prostate Biopsy and Its Efficacy to Reduce Infectious Complications: Systematic Review

Authors: Mohand Yaghi, O. Kehinde

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Background: For the diagnosis of prostate cancer Trans-rectal prostate biopsy (TRPB) is used commonly, the procedure is associated with infective complications. There is evidence that antibiotics (ABx) decrease infective events after TRPB, but different regimens are used. Aim: To systematically review different regimens of prophylactic oral antibiotics in TRPB. Design: Medline, Embase, Clinical trials site, and Cochrane library were searched, experts were consulted about relevant studies. Randomized clinical trials (RCT) conducted in the last twenty years, which investigated different oral antibiotic regimens in TRPB, and compared their efficacy to reduce infectious complications were analyzed. Measurements: Primary outcomes were bacteriuria, urinary tract infection (UTI), fever, bacteremia, sepsis. Secondary outcomes were hospitalization rate, and the prevalence of ABx-resistant bacteria. Results: Nine trials were eligible with 3012 patients. Antibiotics prevented bacteriuria (3.5% vs. 9.88%), UTI (4.46% vs. 9.75%), and hospitalization (0.21% vs. 2.13%) significantly in comparison with placebo or no treatment. No significant difference was found in all outcomes of the review between the single dose regimen and the 3 days. The single dose regimen was as effective as the multiple dose except in Bacteriuria (6.75% vs. 3.25%), and the prevalence of ABx-resistant bacteria (1.57% vs. 0.27%). Quinolones reduced only UTI significantly in comparison with other antibiotics. Lastly, Ciprofloxacin is the best Quinolone to prevent UTI, and hospitalization. Conclusion: it is essential to prescribe prophylactic Antibiotics in TRPB. No conclusive evidence could be claimed about the superiority of the multiple or the 3 days regimens to the single dose regimen. Unexpectedly, ABx-resistant bacteria was identified more often in the single dose cohorts.

Keywords: infection, prostate cancer, sepsis, TRPB

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397 Preparation and Size Control of Sub-100 Nm Pure Nanodrugs

Authors: Jinfeng Zhang, Chun-Sing Lee

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Pure nanodrugs (PNDs) – nanoparticles consisting entirely of drug molecules, have been considered as promising candidates for the next-generation nanodrugs. However, the traditional preparation method via reprecipitation faces critical challenges including low production rates, relatively large particle sizes and batch-to-batch variations. Here, for the first time, we successfully developed a novel, versatile and controllable strategy for preparing PNDs via an anodized aluminium oxide (AAO) template-assisted method. With this approach, we prepared PNDs of an anti-cancer drug (VM-26) with precisely controlled sizes reaching the sub-20 nm range. This template-assisted approach has much higher feasibility for mass production comparing to the conventional reprecipitation method and is beneficial for future clinical translation. The present method is further demonstrated to be easily applicable for a wide range of hydrophobic biomolecules without the need of custom molecular modifications and can be extended for preparing all-in-one nanostructures with different functional agents.

Keywords: drug delivery, pure nanodrugs, size control, template

Procedia PDF Downloads 278
396 Visual Template Detection and Compositional Automatic Regular Expression Generation for Business Invoice Extraction

Authors: Anthony Proschka, Deepak Mishra, Merlyn Ramanan, Zurab Baratashvili

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Small and medium-sized businesses receive over 160 billion invoices every year. Since these documents exhibit many subtle differences in layout and text, extracting structured fields such as sender name, amount, and VAT rate from them automatically is an open research question. In this paper, existing work in template-based document extraction is extended, and a system is devised that is able to reliably extract all required fields for up to 70% of all documents in the data set, more than any other previously reported method. The approaches are described for 1) detecting through visual features which template a given document belongs to, 2) automatically generating extraction rules for a given new template by composing regular expressions from multiple components, and 3) computing confidence scores that indicate the accuracy of the automatic extractions. The system can generate templates with as little as one training sample and only requires the ground truth field values instead of detailed annotations such as bounding boxes that are hard to obtain. The system is deployed and used inside a commercial accounting software.

Keywords: data mining, information retrieval, business, feature extraction, layout, business data processing, document handling, end-user trained information extraction, document archiving, scanned business documents, automated document processing, F1-measure, commercial accounting software

Procedia PDF Downloads 99
395 Glyco-Biosensing as a Novel Tool for Prostate Cancer Early-Stage Diagnosis

Authors: Pavel Damborsky, Martina Zamorova, Jaroslav Katrlik

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Prostate cancer is annually the most common newly diagnosed cancer among men. An extensive number of evidence suggests that traditional serum Prostate-specific antigen (PSA) assay still suffers from a lack of sufficient specificity and sensitivity resulting in vast over-diagnosis and overtreatment. Thus, the early-stage detection of prostate cancer (PCa) plays undisputedly a critical role for successful treatment and improved quality of life. Over the last decade, particular altered glycans have been described that are associated with a range of chronic diseases, including cancer and inflammation. These glycans differences enable a distinction to be made between physiological and pathological state and suggest a valuable biosensing tool for diagnosis and follow-up purposes. Aberrant glycosylation is one of the major characteristics of disease progression. Consequently, the aim of this study was to develop a more reliable tool for early-stage PCa diagnosis employing lectins as glyco-recognition elements. Biosensor and biochip technology putting to use lectin-based glyco-profiling is one of the most promising strategies aimed at providing fast and efficient analysis of glycoproteins. The proof-of-concept experiments based on sandwich assay employing anti-PSA antibody and an aptamer as a capture molecules followed by lectin glycoprofiling were performed. We present a lectin-based biosensing assay for glycoprofiling of serum biomarker PSA using different biosensor and biochip platforms such as label-free surface plasmon resonance (SPR) and microarray with fluorescent label. The results suggest significant differences in interaction of particular lectins with PSA. The antibody-based assay is frequently associated with the sensitivity, reproducibility, and cross-reactivity issues. Aptamers provide remarkable advantages over antibodies due to the nucleic acid origin, stability and no glycosylation. All these data are further step for construction of highly selective, sensitive and reliable sensors for early-stage diagnosis. The experimental set-up also holds promise for the development of comparable assays with other glycosylated disease biomarkers.

Keywords: biomarker, glycosylation, lectin, prostate cancer

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394 Creative Thinking through Mindful Practices: A Business Class Case Study

Authors: Malavika Sundararajan

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This study introduces the use of mindfulness techniques in the classroom to make individuals aware of how the creative thinking process works, resulting in more constructive learning and application. Case observation method was utilized within a classroom setting in a graduate class in the Business School. It entailed, briefing the student participants about the use of a template called the dots and depths map, and having them complete it for themselves, compare it to their team members and reflect on the outputs. Finally, they were debriefed about the use of the template and its value to their learning and creative application process. The major finding is the increase in awareness levels of the participants following the use of the template, leading to a subsequent pursuit of diverse knowledge and acquisition of relevant information and not jumping to solutions directly, which increased their overall creative outputs for the given assignment. The significant value of this study is that it can be applied to any classroom on any subject as a powerful mindfulness tool which increases creative problem solving through constructive knowledge building.

Keywords: connecting dots, mindful awareness, constructive knowledge building, learning creatively

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393 Hyperelastic Constitutive Modelling of the Male Pelvic System to Understand the Prostate Motion, Deformation and Neoplasms Location with the Influence of MRI-TRUS Fusion Biopsy

Authors: Muhammad Qasim, Dolors Puigjaner, Josep Maria López, Joan Herrero, Carme Olivé, Gerard Fortuny

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Computational modeling of the human pelvis using the finite element (FE) method has become extremely important to understand the mechanics of prostate motion and deformation when transrectal ultrasound (TRUS) guided biopsy is performed. The number of reliable and validated hyperelastic constitutive FE models of the male pelvis region is limited, and given models did not precisely describe the anatomical behavior of pelvis organs, mainly of the prostate and its neoplasms location. The motion and deformation of the prostate during TRUS-guided biopsy makes it difficult to know the location of potential lesions in advance. When using this procedure, practitioners can only provide roughly estimations for the lesions locations. Consequently, multiple biopsy samples are required to target one single lesion. In this study, the whole pelvis model (comprised of the rectum, bladder, pelvic muscles, prostate transitional zone (TZ), and peripheral zone (PZ)) is used for the simulation results. An isotropic hyperelastic approach (Signorini model) was used for all the soft tissues except the vesical muscles. The vesical muscles are assumed to have a linear elastic behavior due to the lack of experimental data to determine the constants involved in hyperelastic models. The tissues and organ geometry is taken from the existing literature for 3D meshes. Then the biomechanical parameters were obtained under different testing techniques described in the literature. The acquired parametric values for uniaxial stress/strain data are used in the Signorini model to see the anatomical behavior of the pelvis model. The five mesh nodes in terms of small prostate lesions are selected prior to biopsy and each lesion’s final position is targeted when TRUS probe force of 30 N is applied at the inside rectum wall. Code_Aster open-source software is used for numerical simulations. Moreover, the overall effects of pelvis organ deformation were demonstrated when TRUS–guided biopsy is induced. The deformation of the prostate and neoplasms displacement showed that the appropriate material properties to organs altered the resulting lesion's migration parametrically. As a result, the distance traveled by these lesions ranged between 3.77 and 9.42 mm. The lesion displacement and organ deformation are compared and analyzed with our previous study in which we used linear elastic properties for all pelvic organs. Furthermore, the visual comparison of axial and sagittal slices are also compared, which is taken for Magnetic Resource Imaging (MRI) and TRUS images with our preliminary study.

Keywords: code-aster, magnetic resonance imaging, neoplasms, transrectal ultrasound, TRUS-guided biopsy

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392 The Impact of Using Flattening Filter-Free Energies on Treatment Efficiency for Prostate SBRT

Authors: T. Al-Alawi, N. Shorbaji, E. Rashaidi, M.Alidrisi

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Purpose/Objective(s): The main purpose of this study is to analyze the planning of SBRT treatments for localized prostate cancer with 6FFF and 10FFF energies to see if there is a dosimetric difference between the two energies and how we can increase the plan efficiency and reduce its complexity. Also, to introduce a planning method in our department to treat prostate cancer by utilizing high energy photons without increasing patient toxicity and fulfilled all dosimetric constraints for OAR (an organ at risk). Then toevaluate the target 95% coverage PTV95, V5%, V2%, V1%, low dose volume for OAR (V1Gy, V2Gy, V5Gy), monitor unit (beam-on time), and estimate the values of homogeneity index HI, conformity index CI a Gradient index GI for each treatment plan.Materials/Methods: Two treatment plans were generated for15 patients with localized prostate cancer retrospectively using the CT planning image acquired for radiotherapy purposes. Each plan contains two/three complete arcs with two/three different collimator angle sets. The maximum dose rate available is 1400MU/min for the energy 6FFF and 2400MU/min for 10FFF. So in case, we need to avoid changing the gantry speed during the rotation, we tend to use the third arc in the plan with 6FFF to accommodate the high dose per fraction. The clinical target volume (CTV) consists of the entire prostate for organ-confined disease. The planning target volume (PTV) involves a margin of 5 mm. A 3-mm margin is favored posteriorly. Organs at risk identified and contoured include the rectum, bladder, penile bulb, femoral heads, and small bowel. The prescription dose is to deliver 35Gyin five fractions to the PTV and apply constraints for organ at risk (OAR) derived from those reported in references. Results: In terms of CI=0.99, HI=0.7, and GI= 4.1, it was observed that they are all thesame for both energies 6FFF and 10FFF with no differences, but the total delivered MUs are much less for the 10FFF plans (2907 for 6FFF vs.2468 for 10FFF) and the total delivery time is 124Sc for 6FFF vs. 61Sc for 10FFF beams. There were no dosimetric differences between 6FFF and 10FFF in terms of PTV coverage and mean doses; the mean doses for the bladder, rectum, femoral heads, penile bulb, and small bowel were collected, and they were in favor of the 10FFF. Also, we got lower V1Gy, V2Gy, and V5Gy doses for all OAR with 10FFF plans. Integral dosesID in (Gy. L) were recorded for all OAR, and they were lower with the 10FFF plans. Conclusion: High energy 10FFF has lower treatment time and lower delivered MUs; also, 10FFF showed lower integral and meant doses to organs at risk. In this study, we suggest usinga 10FFF beam for SBRTprostate treatment, which has the advantage of lowering the treatment time and that lead to lessplan complexity with respect to 6FFF beams.

Keywords: FFF beam, SBRT prostate, VMAT, prostate cancer

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391 Easymodel: Web-based Bioinformatics Software for Protein Modeling Based on Modeller

Authors: Alireza Dantism

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Presently, describing the function of a protein sequence is one of the most common problems in biology. Usually, this problem can be facilitated by studying the three-dimensional structure of proteins. In the absence of a protein structure, comparative modeling often provides a useful three-dimensional model of the protein that is dependent on at least one known protein structure. Comparative modeling predicts the three-dimensional structure of a given protein sequence (target) mainly based on its alignment with one or more proteins of known structure (templates). Comparative modeling consists of four main steps 1. Similarity between the target sequence and at least one known template structure 2. Alignment of target sequence and template(s) 3. Build a model based on alignment with the selected template(s). 4. Prediction of model errors 5. Optimization of the built model There are many computer programs and web servers that automate the comparative modeling process. One of the most important advantages of these servers is that it makes comparative modeling available to both experts and non-experts, and they can easily do their own modeling without the need for programming knowledge, but some other experts prefer using programming knowledge and do their modeling manually because by doing this they can maximize the accuracy of their modeling. In this study, a web-based tool has been designed to predict the tertiary structure of proteins using PHP and Python programming languages. This tool is called EasyModel. EasyModel can receive, according to the user's inputs, the desired unknown sequence (which we know as the target) in this study, the protein sequence file (template), etc., which also has a percentage of similarity with the primary sequence, and its third structure Predict the unknown sequence and present the results in the form of graphs and constructed protein files.

Keywords: structural bioinformatics, protein tertiary structure prediction, modeling, comparative modeling, modeller

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390 Quality Assurance Comparison of Map Check 2, Epid, and Gafchromic® EBT3 Film for IMRT Treatment Planning

Authors: Khalid Iqbal, Saima Altaf, M. Akram, Muhammad Abdur Rafaye, Saeed Ahmad Buzdar

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Objective: Verification of patient-specific intensity modulated radiation therapy (IMRT) plans using different 2-D detectors has become increasingly popular due to their ease of use and immediate readout of the results. The purpose of this study was to test and compare various 2-D detectors for dosimetric quality assurance (QA) of intensity-modulated radiotherapy (IMRT) with the vision to find alternative QA methods. Material and Methods: Twenty IMRT patients (12 of brain and 8 of the prostate) were planned on Eclipse treatment planning system using Varian Clinac DHX on both energies 6MV and 15MV. Verification plans of all such patients were also made and delivered to Map check2, EPID (Electronic portal imaging device) and Gafchromic EBT3. Gamma index analyses were performed using different criteria to evaluate and compare the dosimetric results. Results: Statistical analysis shows the passing rate of 99.55%, 97.23% and 92.9% for 6MV and 99.53%, 98.3% and 94.85% for 15 MV energy using a criteria of ±5% of 3mm, ±3% of 3mm and ±3% of 2mm respectively for brain, whereas using ±5% of 3mm and ±3% of 3mm gamma evaluation criteria, the passing rate is 94.55% and 90.45% for 6MV and 95.25%9 and 95% for 15 MV energy for the case of prostate using EBT3 film. Map check 2 results shows the passing rates of 98.17%, 97.68% and 86.78% for 6MV energy and 94.87%,97.46% and 88.31% for 15 MV energy respectively for brain using a criteria of ±5% of 3mm, ±3% of 3mm and ±3% of 2mm, whereas using ±5% of 3mm and ±3% of 3mm gamma evaluation criteria gives the passing rate of 97.7% and 96.4% for 6MV and 98.75%9 and 98.05% for 15 MV energy for the case of prostate. EPID 6 MV and gamma analysis shows the passing rate of 99.56%, 98.63% and 98.4% for the brain, 100% and 99.9% for prostate using the same criteria as for map check 2 and EBT 3 film. Conclusion: The results demonstrate excellent passing rates were obtained for all dosimeter when compared with the planar dose distributions for 6 MV IMRT fields as well as for 15 MV. EPID results are better than EBT3 films and map check 2 because it is likely that part of this difference is real, and part is due to manhandling and different treatment set up verification which contributes dose distribution difference. Overall all three dosimeter exhibits results within limits according to AAPM report.120.

Keywords: gafchromic EBT3, radiochromic film dosimetry, IMRT verification, EPID

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389 ALDH1A1 as a Cancer Stem Cell Marker: Value of Immunohistochemical Expression in Benign Prostatic Hyperplasia, Prostatic Intraepithelial Neoplasia, and Prostatic Adenocarcinoma

Authors: H. M. Abdelmoneim, N. A. Babtain, A. S. Barhamain, A. Z. Kufiah, A. S. Malibari, S. F. Munassar, R. S. Rawa

Abstract:

Introduction: Prostate cancer is one of the most common causes of morbidity and mortality in men in developed countries. Cancer Stem Cells (CSCs) could be responsible for the progression and relapse of cancer. Therefore, CSCs markers could provide a prognostic strategy for human malignancies. Aldehyde dehydrogenase 1A1 (ALDH1A1) activity has been shown to be associated with tumorigenesis and proposed to represent a functional marker for tumor initiating cells in various tumor types including prostate cancer. Material & Methods: We analyzed the immunohistochemical expression of ALDH1A1 in benign prostatic hyperplasia (BPH), prostatic intraepithelial neoplasia (PIN) and prostatic adenocarcinoma and assessed their significant correlations in 50 TURP sections. They were microscopically interpreted and the results were correlated with histopathological types and tumor grade. Results: In different prostatic histopathological lesions we found that ALDH1A1 expression was low in BPH (13.3%) and PIN (6.7%) and then its expression increased with prostatic adenocarcinoma (40%), and this was statistically highly significant (P value = 0.02). However, in different grades of prostatic adenocarcinoma we found that the higher the Gleason grade the higher the expression for ALDH1A1 and this was statistically significant (P value = 0.02). We compared the expression of ALDH1A1 in PIN and prostatic adenocarcinoma. ALDH1A1 expression was decreased in PIN and highly expressed in prostatic adenocarcinoma and this was statistically significant (P value = 0.04). Conclusion: Increasing ALDH1A1 expression is correlated with aggressive behavior of the tumor. Immunohistochemical expression of ALDH1A1 might provide a potential approach to study tumorigenesis and progression of primary prostate carcinoma.

Keywords: ALDH1A1, BPH, PIN, prostatic adenocarcinoma

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388 Thulium Laser Vaporisation and Enucleation of Prostate in Patients on Anticoagulants and Antiplatelet Agents

Authors: Abdul Fatah, Naveenchandra Acharya, Vamshi Krishna, T. Shivaprasad, Ramesh Ramayya

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Background: Significant number of patients with bladder outlet obstruction due to BPH are on anti-platelets and anticoagulants. Prostate surgery in this group of patients either in the form of TURP or Open prostatectomy is associated with increased risk of bleeding complications requiring transfusions, packing of the prostatic fossa or ligation or embolization of internal iliac arteries. Withholding of antiplatelets and anticoagulants may be associated with cardiac and other complications. Efficacy of Thulium Laser in the above group of patients was evaluated in terms of peri-operative, postoperative and delayed bleeding complications as well as cardiac events in peri-operative and immediate postoperative period. Methods: 217 patients with a mean age of 68.8 years were enrolled between March 2009 and March 2013 (36 months), and treated for BPH with ThuLEP. Every patient was evaluated at base line according to: Digital Rectal Examination (DRE), prostate volume, Post-Voided volume (PVR), International Prostate Symptoms Score (I-PSS), PSA values, urine analysis and urine culture, uroflowmetry. The post operative complications in the form of drop in hemoglobin level, transfusion rates, post –operative cardiac events within a period of 30 days, delayed hematuria and events like deep vein thrombosis and pulmonary embolism were noted. Results: Our data showed a better post-operative outcome in terms of, postoperative bleeding requiring intervention 7 (3.2%), transfusion rate 4 (1.8%) and cardiac events within a period of 30 days 4(1.8%), delayed hematuria within 6 months 2(0.9 %) compared other series of prostatectomies. Conclusion: The thulium LASER prostatectomy is a safe and effective option for patients with cardiac comorbidties and those patients who are on antiplatelet agents and anticoagulants. The complication rate is less as compared to larger series reported with open and transurethral prostatectomies.

Keywords: thulium laser, prostatectomy, antiplatelet agents, bleeding

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387 SnSₓ, Cu₂ZnSnS₄ Nanostructured Thin Layers for Thin-Film Solar Cells

Authors: Elena A. Outkina, Marina V. Meledina, Aliaksandr A. Khodin

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Nanostructured thin films of SnSₓ, Cu₂ZnSnS₄ (CZTS) semiconductors were fabricated by chemical processing to produce thin-film photoactive layers for photocells as a prospective lowest-cost and environment-friendly alternative to Si, Cu(In, Ga)Se₂, and other traditional solar cells materials. To produce SnSₓ layers, the modified successive ionic layer adsorption and reaction (SILAR) technique were investigated, including successive cyclic dipping into Na₂S solution and SnCl₂, NaCl, triethanolamine solution. To fabricate CZTS layers, the cyclic dipping into CuSO₄ with ZnSO₄, SnCl₂, and Na₂S solutions was used with intermediate rinsing in distilled water. The nano-template aluminum/alumina substrate was used to control deposition processes. Micromorphology and optical characteristics of the fabricated layers have been investigated. Analysis of 2D-like layers deposition features using nano-template substrate is presented, including the effect of nanotips in a template on surface charge redistribution and transport.

Keywords: kesterite, nanotemplate, SILAR, solar cell, tin sulphide

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386 Inhibitory Effect of P2Y1R Agonist 1-Indolinoalkyl 2-Phenolic Derivative on Prostate Cancer Cell Proliferation via the MAPK Signalling

Authors: Hien Thi Thu Le, Nuno Rafael Candeias, Olli Yli-Harja, Meenakshisundaram Kandhavelu

Abstract:

Purinergic receptor 1 (P2Y1R) is the potential therapeutic target for inducing prostate cancer (PCa) cell death. Recently, 1-indolinoalkyl 2-phenolic derivative, HIC, was identified as a P2Y1R agonist that increases apoptosis and inhibits cell proliferation of PCa. However, the biological effects of HIC have not been extensively studied at the molecular level. In the present study, we have investigated the anticancer effects of HIC and the molecular mechanisms underlying in PCa cells. Half maximal inhibitory concentration (IC₅₀) of HIC was measured as 15.98 μM and 15.64 μM for DU145 and PC3 cells, respectively. In addition, we found that HIC inhibited cell growth and metastasis of PC3 and DU145 cells colonies, spheroid areas, and migrated cells. RNA seq analysis revealed significant changes of over 3000 genes (p value < 0.05) upon HIC treatment in PC3 and DU145 cells. Genes involved in DNA damage, apoptosis, cell cycle arrest at G1/S phase were modulated by HIC treatment. MAPK and NF-κB protein array revealed the increased expression of ERK1/2, JNK1/2, p53 phosphorylation, and p53 protein. ERK1/2 and JNK1/2 activations are known to increase the stabilization of p53, a tumor suppressor protein, which is required to arrest the cell cycle at G1/S phase and cause cell death of PCa cells. Overall, our results suggest that HIC can serve as a multi-dimensional chemotherapeutic agent possessing strong cytotoxic, anti-cancer, and anti-metastasis against PCa growth.

Keywords: prostate cancer, P2Y1 receptor, apoptosis, metastasis

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385 Detection of JC Virus DNA and T-Ag Expression in a Subpopulation of Tunisian Colorectal Carcinomas

Authors: Wafa Toumi, Alessandro Ripalti, Luigi Ricciardiello, Dalila Gargouri, Jamel Kharrat, Abderraouf Cherif, Ahmed Bouhafa, Slim Jarboui, Mohamed Zili, Ridha Khelifa

Abstract:

Background & aims: Colorectal cancer (CRC) is one of the most common malignancies throughout the world. Several risk factors, both genetic and environmental, including viral infections, have been linked to colorectal carcinogenesis. A few studies report the detection of human polyomavirus JC (JCV) DNA and transformation antigen (T-Ag) in a fraction of the colorectal tumors studied and suggest an association of this virus with CRC. In order to investigate whether such an association of JCV with CRC will hold in a different epidemiological setting, we looked for the presence of JCV DNA and T-Ag expression in a group of Tunisian CRC patients. Methods: Fresh colorectal mucosa biopsies were obtained from 17 healthy volunteers and from both colorectal tumors and adjacent normal tissues of 47 CRC patients. DNA was extracted from fresh biopsies or from formalin-fixed, paraffin-embedded tissue sections using the Invitrogen Purelink Genomic DNA mini Kit. A simple PCR and a nested PCR were used to amplify a region of the T-Ag gene. The obtained PCR products revealed a 154 bp and a 98 bp bands, respectively. Specificity was confirmed by sequencing of the PCR products. T-Ag expression was determined by immunohistochemical staining using a mouse monoclonal antibody (clone PAb416) directed against SV40 T-Ag that cross reacts with JCV T-Ag. Results: JCV DNA was found in 12 (25%) and 22 (46%) of the CRC tumors by simple PCR and by nested PCR, respectively. All paired adjacent normal mucosa biopsies were negative for viral DNA. Sequencing of the DNA amplicons obtained confirmed the authenticity of T-Ag sequences. Immunohistochemical staining showed nuclear T-Ag expression in all 22 JCV DNA- positive samples and in 3 additional tumor samples which appeared DNA-negative by PCR. Conclusions: These results suggest an association of JCV with a subpopulation of Tunisian colorectal tumors.

Keywords: colorectal cancer, immunohistochemistry, Polyomavirus JC, PCR

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384 Vertically Grown P–Type ZnO Nanorod on Ag Thin Film

Authors: Jihyun Park, Tae Il Lee, Jae-Min Myoung

Abstract:

A Silver (Ag) thin film is introduced as a template and doping source for vertically aligned p–type ZnO nanorods. ZnO nanorods were grown using a ammonium hydroxide based hydrothermal process. During the hydrothermal process, the Ag thin film was dissolved to generate Ag ions in the solution. The Ag ions can contribute to doping in the wurzite structure of ZnO and the (111) grain of Ag thin film can be the epitaxial temporal template for the (0001) plane of ZnO. Hence, Ag–doped p–type ZnO nanorods were successfully grown on the substrate, which can be an electrode or semiconductor for the device application. To demonstrate the potentials of this idea, p–n diode was fabricated and its electrical characteristics were demonstrated.

Keywords: hydrothermal process, Ag–doped ZnO nanorods, p–type ZnO

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383 Profiling of Apoptotic Protein Expressions after Trabectedin Treatment in Human Prostate Cancer Cell Line PC-3 by Protein Array Technology

Authors: Harika Atmaca, Emir Bozkurt, Latife Merve Oktay, Selim Uzunoglu, Ruchan Uslu, Burçak Karaca

Abstract:

Microarrays have been developed for highly parallel enzyme-linked immunosorbent assay (ELISA) applications. The most common protein arrays are produced by using multiple monoclonal antibodies, since they are robust molecules which can be easily handled and immobilized by standard procedures without loss of activity. Protein expression profiling with protein array technology allows simultaneous analysis of the protein expression pattern of a large number of proteins. Trabectedin, a tetrahydroisoquinoline alkaloid derived from a Caribbean tunicate, Ecteinascidia turbinata, has been shown to have antitumor effects. Here, we used a novel proteomic approach to explore the mechanism of action of trabectedin in prostate cancer cell line PC-3 by apoptosis antibody microarray. XTT cell proliferation kit and Cell Death Detection Elisa Plus Kit (Roche) was used for measuring cytotoxicity and apoptosis. Human Apoptosis Protein Array (R&D Systems) which consists of 35 apoptosis related proteins was used to assess the omic protein expression pattern. Trabectedin induced cytotoxicity and apoptosis in prostate cancer cells in a time and concentration-dependent manner. The expression levels of the death receptor pathway molecules, TRAIL-R1/DR4, TRAIL R2/DR5, TNF R1/TNFRSF1A, FADD were significantly increased by 4.0-, 21.0-, 4.20- and 11.5-fold by trabectedin treatment in PC-3 cells. Moreover, mitochondrial pathway related pro-apoptotic proteins Bax, Bad, Cytochrome c, and Cleaved Caspase-3 expressions were induced by 2.68-, 2.07-, 2.8-, and 4.5-fold and the expression levels of anti-apoptotic proteins Bcl-2 and Bcl-XL were reduced by 3.5- and 5.2-fold in PC-3 cells. Proteomic (antibody microarray) analysis suggests that the mechanism of action of trabectedin may be exerted via the induction of both intrinsic and extrinsic apoptotic pathways. The antibody microarray platform can be utilised to explore the molecular mechanism of action of novel anticancer agents.

Keywords: trabectedin, prostate cancer, omic protein expression profile, apoptosis

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382 Cost-Effective Soft Lithography of Organic Semiconductors in Organic Field-Effect Transistors (OFETs)

Authors: Tae Kyu An

Abstract:

We demonstrate repurposing linear micropatterns on the CD as a master mold to fabricate TIPS-PEN microwires. From the micropatterns on CDs, we replicated polyurethane acrylate (PUA) templates which are robust and flexible until submicrometer scale patterns. Subsequently, 1.5 μm TIPS-PEN microwires separated by 1.5 μm were grown. Using crystal analysis tools with polarized optical microscopy and X-ray diffraction measurement, it was revealed that each TIPS-PEN microwires are highly crystalline and uniform compared to spin-coated films. It is attributed to the template-guided growth of TIPS-PEN crystals along the linear template, thus the OFETs comprised of TIPS-PEN microwires displayed the high field-effect mobility.

Keywords: compact disk, macro patterning, OFET, soft lithography

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381 A Biometric Template Security Approach to Fingerprints Based on Polynomial Transformations

Authors: Ramon Santana

Abstract:

The use of biometric identifiers in the field of information security, access control to resources, authentication in ATMs and banking among others, are of great concern because of the safety of biometric data. In the general architecture of a biometric system have been detected eight vulnerabilities, six of them allow obtaining minutiae template in plain text. The main consequence of obtaining minutia templates is the loss of biometric identifier for life. To mitigate these vulnerabilities several models to protect minutiae templates have been proposed. Several vulnerabilities in the cryptographic security of these models allow to obtain biometric data in plain text. In order to increase the cryptographic security and ease of reversibility, a minutiae templates protection model is proposed. The model aims to make the cryptographic protection and facilitate the reversibility of data using two levels of security. The first level of security is the data transformation level. In this level generates invariant data to rotation and translation, further transformation is irreversible. The second level of security is the evaluation level, where the encryption key is generated and data is evaluated using a defined evaluation function. The model is aimed at mitigating known vulnerabilities of the proposed models, basing its security on the impossibility of the polynomial reconstruction.

Keywords: fingerprint, template protection, bio-cryptography, minutiae protection

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380 Synthetic Dermal Template Use in the Reconstruction of a Chronic Scalp Wound

Authors: Stephanie Cornish

Abstract:

The use of synthetic dermal templates, also known as dermal matrices, such as PolyNovo® Biodegradable Temporising Matrix (BTM), has been well established in the reconstruction of acute wounds with a full thickness defect of the skin. Its use has become common place in the treatment of full thickness burns and is not unfamiliar in the realm of necrotising fasciitis, free flap donor site reconstruction, and the management of acute traumatic wounds. However, the use of dermal templates for more chronic wounds is rare. The authors present the successful use of BTM in the reconstruction of a chronic scalp wound following the excision of a malignancy and multiple previous failed attempts at repair, thus demonstrating the potential for an increased scope of use.

Keywords: dermal template, BTM, chronic, scalp wound, reconstruction

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379 Antiproliferative and Apoptotic Effects of an Enantiomerically Pure β-Dipeptide Derivative through PI3K/Akt-Dependent and -Independent Pathways in Human Hormone-Refractory Prostate Cancer Cells

Authors: Mei-Ling Chan, Jin-Ming Wu, Konstantin V. Kudryavtsev, Jih-Hwa Guh

Abstract:

Prostate cancer is one of the most common malignant disease in men. KUD983 is an enantiomerically pure β-dipeptide derivative, which may have anti-cancer effects. In the present study, KUD983 exhibits powerful activity against hormone-refractory prostate cancer (HRPC) PC-3 and DU145 cells. The IC50 values of KUD983 in PC-3 and DU145 cells are 0.56±0.07M and 0.50±0.04 M respectively. KUD983 induced G1 arrest of the cell cycle and subsequent apoptosis associated with the down-regulation of several related proteins including cyclin D1, cyclin E and Cdk4, and the de-phosphorylation of RB. The protein expressions of nuclear and total c-Myc protein, which was able to regulate the expression of both cyclin D1 and cyclin E, were significantly suppressed by KUD983. Phosphoinositide 3-kinase (PI3K)/Akt/mammalian target of rapamycin (mTOR) is an important signaling pathway that influences the energy metabolism, cell cycle, proliferation, survival and apoptosis of cells, and is associated with numerous other signaling pathways. The Western Blot data revealed that KUD983 inhibited PI3K/Akt and mTOR/p70S6K/4E-BP1 pathways. The transient transfection of constitutively active myristylated Akt (myr-Akt) cDNA significantly reversed KUD983-induced caspase activation but did not abolish the suppression of mTOR/p70S6K/4E-BP1 signaling cascade indicating the presence of both Akt-dependent and -independent pathways. Moreover, KUD983-induced effect was collaborated with the down-regulation of anti-apoptotic Bcl-2 members (e.g., Bcl-2, and Mcl-1) and IAP family members (e.g., survivin). Furthermore, KUD983 induced autophagic cell death using confocal microscopic examination, investigating the level of conversion of LC3-I to LC3-II and flow cytometric detection of AVO-positive cells. Taken together, the data suggest that KUD983 is an anticancer β-dipeptide against HRPCs through the inhibition of cell proliferation and induction of apoptotic and autophagic cell death. The suppression of signaling pathways mediated by c-Myc, PI3K/Akt and mTOR/p70S6K/4E-BP1 and the collaboration with down-regulation of Mcl-1 and survivin may indicate the mechanism of KUD983 against HRPC.

Keywords: β-dipeptide, hormone-refractory prostate cancer, mTOR, PI3K/Akt

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378 Multi-Template Molecularly Imprinted Polymer: Synthesis, Characterization and Removal of Selected Acidic Pharmaceuticals from Wastewater

Authors: Lawrence Mzukisi Madikizela, Luke Chimuka

Abstract:

Removal of organics from wastewater offers a better water quality, therefore, the purpose of this work was to investigate the use of molecularly imprinted polymer (MIP) for the elimination of selected organics from water. A multi-template MIP for the adsorption of naproxen, ibuprofen and diclofenac was synthesized using a bulk polymerization method. A MIP was synthesized at 70°C by employing 2-vinylpyridine, ethylene glycol dimethacrylate, toluene and 1,1’-azobis-(cyclohexanecarbonitrile) as functional monomer, cross-linker, porogen and initiator, respectively. Thermogravimetric characterization indicated that the polymer backbone collapses at 250°C and scanning electron microscopy revealed the porous and roughness nature of the MIP after elution of templates. The performance of the MIP in aqueous solutions was evaluated by optimizing several adsorption parameters. The optimized adsorption conditions were 50 mg of MIP, extraction time of 10 min, a sample pH of 4.6 and the initial concentration of 30 mg/L. The imprinting factors obtained for naproxen, ibuprofen and diclofenac were 1.25, 1.42, and 2.01, respectively. The order of selectivity for the MIP was; diclofenac > ibuprofen > naproxen. MIP showed great swelling in water with an initial swelling rate of 2.62 g/(g min). The synthesized MIP proved to be able to adsorb naproxen, ibuprofen and diclofenac from contaminated deionized water, wastewater influent and effluent.

Keywords: adsorption, molecularly imprinted polymer, multi template, pharmaceuticals

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377 Ultra-Sensitive Point-Of-Care Detection of PSA Using an Enzyme- and Equipment-Free Microfluidic Platform

Authors: Ying Li, Rui Hu, Shizhen Chen, Xin Zhou, Yunhuang Yang

Abstract:

Prostate cancer is one of the leading causes of cancer-related death among men. Prostate-specific antigen (PSA), a specific product of prostatic epithelial cells, is an important indicator of prostate cancer. Though PSA is not a specific serum biomarker for the screening of prostate cancer, it is recognized as an indicator for prostate cancer recurrence and response to therapy for patient’s post-prostatectomy. Since radical prostatectomy eliminates the source of PSA production, serum PSA levels fall below 50 pg/mL, and may be below the detection limit of clinical immunoassays (current clinical immunoassay lower limit of detection is around 10 pg/mL). Many clinical studies have shown that intervention at low PSA levels was able to improve patient outcomes significantly. Therefore, ultra-sensitive and precise assays that can accurately quantify extremely low levels of PSA (below 1-10 pg/mL) will facilitate the assessment of patients for the possibility of early adjuvant or salvage treatment. Currently, the commercially available ultra-sensitive ELISA kit (not used clinically) can only reach a detection limit of 3-10 pg/mL. Other platforms developed by different research groups could achieve a detection limit as low as 0.33 pg/mL, but they relied on sophisticated instruments to get the final readout. Herein we report a microfluidic platform for point-of-care (POC) detection of PSA with a detection limit of 0.5 pg/mL and without the assistance of any equipment. This platform is based on a previously reported volumetric-bar-chart chip (V-Chip), which applies platinum nanoparticles (PtNPs) as the ELISA probe to convert the biomarker concentration to the volume of oxygen gas that further pushes the red ink to form a visualized bar-chart. The length of each bar is used to quantify the biomarker concentration of each sample. We devised a long reading channel V-Chip (LV-Chip) in this work to achieve a wide detection window. In addition, LV-Chip employed a unique enzyme-free ELISA probe that enriched PtNPs significantly and owned 500-fold enhanced catalytic ability over that of previous V-Chip, resulting in a significantly improved detection limit. LV-Chip is able to complete a PSA assay for five samples in 20 min. The device was applied to detect PSA in 50 patient serum samples, and the on-chip results demonstrated good correlation with conventional immunoassay. In addition, the PSA levels in finger-prick whole blood samples from healthy volunteers were successfully measured on the device. This completely stand-alone LV-Chip platform enables convenient POC testing for patient follow-up in the physician’s office and is also useful in resource-constrained settings.

Keywords: point-of-care detection, microfluidics, PSA, ultra-sensitive

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376 Identifying Common Behavioural Traits of Lone-Wolves in Recent Terrorist Attacks in Europe

Authors: Khaled M. Khan, Armstrong Nhlabatsi

Abstract:

This article attempts to analyse behavioural traits of lone-wolves who struck and killed innocents in six different attacks in Europe in last nine months. The main objective of this study is to develop a profiling template in order to capture commonality of characteristics of these attackers. This study tries to understand the homogeneity of lone-wolves in terms of their social background and state of mind. The commonality among them can possibly be used to build a profiling template that could help detecting vulnerable persons who are prone to be self-radicalised or radicalised by someone else. The result of this study provides us an understanding of their commonality in terms of their state of mind and social characteristics.

Keywords: behavioral pattern, terrorism, profiling, commonality

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375 Single Cell Analysis of Circulating Monocytes in Prostate Cancer Patients

Authors: Leander Van Neste, Kirk Wojno

Abstract:

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

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

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