Search results for: live cells imaging

Authors: Manal Farea, Adam Husein, Ahmad S. Halim, Zurairah Berahim, Nurul A. Abdullah, Khairani I. Mokhtar, Kasmawati Mokhtar

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Endodontic furcal perforation remains both an endodontic and a periodontal problem. Regeneration of cementum is very essential for the perforation repair. The aim of this study was to investigate the role of Hertwig's epithelial root sheath (HERS) cells on the cementogenic differentiation of stem cells derived from human exfoliated deciduous teeth (SHED) in the presence of chitosan scaffold-TGFβ1. HERS cells were isolated and characterized then co-cultured with SHED with/without chitosan scaffold-TGFβ1. SHED proliferation was assessed by PrestoBlue. Alkaline phosphatase activity, mineralization behaviour and gene/protein expression of cemento/osteoblast phenotype of SHED were evaluated. Results of the present study showed that HERS cells in association with chitosan-TGFβ1 enhanced proliferation and cemento/osteogenic differentiation of SHED. Our novel co-culture system confirmed the potential effect of HERS cells to stimulate the differentiation of SHED along the cementoblastic lineage which was triggered in the presence of chitosan-TGFβ1. This approach possesses a novel therapeutic strategy for future endodontic perforation and periodontitis.

Keywords: cementogenesis, co-culture system, HERS, SHED

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Authors: Vinaya Kambappa, Chinnadurai Mani, Komaraiah Palle

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

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

Procedia PDF Downloads 187

Authors: Lucian Mocan, Flaviu Tabaran, Teodora Mocan, Cristian Matea, Cornel Iancu

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We present method of Gold Nanoparticle enhanced laser thermal ablation of HepG2 cells (Human hepatocellular liver carcinoma cell line), based on a simple gold nanoparticle carrier system, such as serum albumin (BSA), and demonstrate its selective therapeutic efficacy. Hyperspectral, contrast phase, and confocal microscopy combined immunochemical staining were used to demonstrate the selective internalization of HSA-GNPs via Gp60 receptors and the caveolin-mediated endocytosis inside HepG2 cells. We examined the ability of laser-activated carbon nanotubes to induce Hsp70 expression using confocal microscopy. Hep G2 cells heat-shocked (laser activated BSA-GNPs) to 42°C demonstrated an up-regulation of Hsp70 compared with control cells (BSA-GNPs treated cells without laser), which showed no detectable constitutive expression of Hsp70. We observed a time-dependent induction in Hsp70 expression in Hep G2 treated with BSA-GNPs and LASER irradiated. The post-irradiation apoptotic rate of HepG2 cells treated with HSA-GNPs ranged from 88.24% (for 50 mg/L) at 60 seconds, while at 30 minute the rate increased to 92.34% (50 mg/L). These unique results may represent a major step in liver cancer treatment using nanolocalized thermal ablation by laser heating.

Keywords: gold nanoparticles, liver cancer, albumin, laser irradiation

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Authors: Aysegul Alyamac, Sukru Gulec

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Ankaferd Blood Stopper (ABS) is a plant extract used to stop bleeding caused by injuries and surgical interventions. ABS also involved in wound healing of intestinal mucosal damage due to oxidative stress and inflammation. Inflammatory Bowel Disease (IBD) is a common chronic disorder of the gastrointestinal tract that causes abdominal pain, diarrhea, and gastrointestinal bleeding, and increases the risk of colon cancer. Inflammation is an essential factor in the development of IBD. The various studies have been performed about the physiological effects of ABS; however, ABS dependent mechanism on colonic inflammation has not been elucidated. Thus, the protective effect of ABS on colonic inflammation was investigated in this study. The Caco-2 and RAW 264.7 murine macrophage cells were used as a model of in vitro colonic inflammation. RAW 264.7 cells were treated with lipopolysaccharide (LPS) for 12 hours to induce the inflammation, and a conditional medium was obtained. Caco-2 cells were treated with 15 µl/ml ABS for 4 hours, then incubated with conditional medium and the cells also were incubated with 15 µl/ml ABS and conditional medium together for 4 hours. Tumor necrosis factor alpha (TNF-α) protein levels were targeted in testing inflammatory condition and its level was significantly increased (25 fold, p<0.001) compared to the control group by using Enzyme-Linked Immunosorbent Assay (ELISA) method. The COX-2 mRNA level was used as a marker gene to show the possible anti-inflammatory effect of ABS in Caco-2 cells. RAW cells-derived conditional medium significantly (3.3 fold, p<0.001) induced cyclooxygenase-2 (COX-2) mRNA levels in Caco-2 cells. The pretreatment of Caco-2 cells caused a significant decrease (3.3 fold, p<0.001) in COX-2 mRNA levels relative to conditional medium given group. Furthermore, COX-2 mRNA level was significantly reduced (4,7 fold, p<0.001) in ABS and conditional medium treated group. These results suggest that ABS might have an anti-inflammatory effect in vitro.

Keywords: Ankaferd blood stopper, CaCo-2, colonic inflammation, RAW 264.7

Procedia PDF Downloads 146

Authors: Kevser Dincer, Basma Waisi, M. Ozan Ozdemir, Ugur Pasaogullari, Jeffrey McCutcheon

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Nanofibers are defined as fibers with diameters less than 100 nanometers. They can be produced by interfacial polymerization, electrospinning and electrostatic spinning. In this study, behaviours of activated carbon nano fiber (ACNF), carbon nano-fiber (CNF), Polyacrylonitrile/carbon nanotube (PAN/CNT), Polyvinyl alcohol/nano silver (PVA/Ag) in PEM fuel cells are investigated experimentally. This material was used as gas diffusion layer (GDL) in PEM fuel cells. When the performances of these cells are compared to each other at 5x5 cm2 cell, it is found that the PVA/Ag exhibits the best performance among all. In this work, nano fiber and nano fiber/nano particles electrical conductivities have been studied to understand their effects on PEM fuel cell performance. According to the experimental results, the maximum electrical conductivity performance of the fuel cell with nanofiber was found to be at PVA/Ag. The electrical conductivities of CNF, ACNF, PAN/CNT are lower for PEM. The resistance of cell with PVA/Ag is lower than the resistance of cell with PAN/CNT, ACNF, CNF.

Keywords: proton exchange membrane fuel cells, electrospinning, carbon nano fiber, activate carbon nano-fiber, PVA fiber, PAN fiber, carbon nanotube, nano particle nanocomposites

Procedia PDF Downloads 392

Authors: M. Zehtabian, Z. Molaiemanesh, Z. Shafahi, M. Papie, M. Zahraie Moghaddam, M. Mehralizadeh, M. R. Vahidi, S. Sina

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The introduction of CT scans has been a great improvement in diagnosis of different diseases. However, this imaging modality can expose the patients to cumulative radiation doses which may increase the risks of some health problems like cancer. In this study, the dose delivered to different organs in lumbar-abdominal imaging was measured by putting the TLD-100, and TLD-100H chips inside the Alderson Rando phantom. The lumbar-abdominal image of the phantom was obtained, while TLD chips were inside the holes of the phantom. According to the results obtained in this study using TLD-100 chips, the average dose received by liver, bladder, rectum, kidneys, and uterus were found to be 12.9 mSv, 8.9 mSv, 10.1 mSv, 11.0 mSv, 11.2 mSv, and 10.5 mSv respectively, while the measurements performed by TLD-100H show that the average dose to liver, bladder, rectum, kidneys, and uterus were found to be 12.4 mSv, 9.2 mSv, 9.5 mSv, 10.5 mSv, 10.7 mSv, and 9.9 mSv respectively. The results of this study indicates that the dose measured by the TLD-100H chips are in close agreement with those obtained by TLD-100.

Keywords: CT scan, dose, TLD-100, diagnosis

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Authors: Thomas E. Portegys

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An animal behavior problem is presented in the form of a nest-building task that involves two cooperating virtual birds, a male and female. The female builds a nest into which she lays an egg. The male's job is to forage in a forest for food for both himself and the female. In addition, the male must fetch stones from a nearby desert for the female to use as nesting material. The task is completed when the nest is built, and an egg is laid in it. A goal-seeking neural network and a recurrent neural network were trained and tested with little success. The goal-seeking network was then enhanced with “place cells”, allowing the birds to spatially navigate the world, building the nest while keeping themselves fed. Place cells are neurons in the hippocampus that map space.

Keywords: artificial animal intelligence, artificial life, goal-seeking neural network, nest-building, place cells, spatial navigation

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Authors: Patarapan Odton

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This study investigates demographic and socio-economic factors of breastfeeding practice, including exclusively breastfeeding among children in Thailand using the Multiple Indicator Cluster Survey (MICS3 and MICS4). Logistic regression models were used to examine the determinants of initial breastfeeding, exclusively breastfeeding, and predominant breastfeeding, using data from women and children section of the survey. For initial breastfeeding, women live in rural area were more likely to start breastfeeding within one day of birth rather than who live in urban area in both round of the surveys. In year 2012, there were significantly higher probabilities of women in rural area started breastfeeding within one hour of birth compare to urban area. Women in southern Thailand have higher probabilities of start breastfeeding within one hour and one day than women in Bangkok and central region. During the year 2005-2006, children aged less than 5 years old lived in rural area have been breastfed higher than children in urban area. Children live in the northeast region were more likely to have been breastfed than the other regions. Only the second wealth quintile group was significant higher probability of ever been breastfed than the poorest group. The findings in the second round of the survey are different from the year 2005-06. In 2012, there was no difference in probability of ever been breastfed among children live in urban and rural area, children in Bangkok and central region were less probability of ever been breastfed than the others.

Keywords: Breastfeeding, Exclusive Breastfeeding, Predominant Breastfeeding, Urban-Rural Difference

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Authors: Chia-Yu Lin, Yu-Hsiang Shen, Cing-Ciao Ke, Chia-Hao Chang, Fan-Pin Tseng, Yu-Ching Ni, Sheng-Pin Tseng

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Tomosynthesis is an imaging system that generates a 3D image by scanning in a limited angular range. It could provide more depth information than traditional 2D X-ray single projection. Radiation dose in tomosynthesis is less than computed tomography (CT). Because of limited angular range scanning, there are many properties depending on scanning direction. Therefore, non-coplanar imaging technique was developed to improve image quality in traditional tomosynthesis. The purpose of this study was to establish the non-coplanar imaging technique of tomosynthesis system and evaluate this technique by the reconstructed image. INER prototype tomosynthesis system contains an X-ray tube, a flat panel detector, and a motion machine. This system could move X-ray tube in multiple directions during the acquisition. In this study, we investigated three different imaging techniques that were 2D X-ray single projection, traditional tomosynthesis, and non-coplanar tomosynthesis. An anthropopathic chest phantom was used to evaluate the image quality. It contained three different size lesions (3 mm, 5 mm and, 8 mm diameter). The traditional tomosynthesis acquired 61 projections over a 30 degrees angular range in one scanning direction. The non-coplanar tomosynthesis acquired 62 projections over 30 degrees angular range in two scanning directions. A 3D image was reconstructed by iterative image reconstruction algorithm (ML-EM). Our qualitative method was to evaluate artifacts in tomosynthesis reconstructed image. The quantitative method was used to calculate a peak-to-valley ratio (PVR) that means the intensity ratio of the lesion to the background. We used PVRs to evaluate the contrast of lesions. The qualitative results showed that in the reconstructed image of non-coplanar scanning, anatomic structures of chest and lesions could be identified clearly and no significant artifacts of scanning direction dependent could be discovered. In 2D X-ray single projection, anatomic structures overlapped and lesions could not be discovered. In traditional tomosynthesis image, anatomic structures and lesions could be identified clearly, but there were many artifacts of scanning direction dependent. The quantitative results of PVRs show that there were no significant differences between non-coplanar tomosynthesis and traditional tomosynthesis. The PVRs of the non-coplanar technique were slightly higher than traditional technique in 5 mm and 8 mm lesions. In non-coplanar tomosynthesis, artifacts of scanning direction dependent could be reduced and PVRs of lesions were not decreased. The reconstructed image was more isotropic uniformity in non-coplanar tomosynthesis than in traditional tomosynthesis. In the future, scan strategy and scan time will be the challenges of non-coplanar imaging technique.

Keywords: image reconstruction, non-coplanar imaging technique, tomosynthesis, X-ray imaging

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Authors: Akhmadjon Sultanov, Eun-Ho Lee, Hye-Jin Park, Young-Je Cho

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This study tested the anti-inflammatory effect of wild indigo (Baptisia tinctoria) root in Raw 264.7 cells. We prepared two extracts of B. tinctoria; one in water and the other in 50% ethanol. Then we evaluated the toxicities of the B. tinctoria root extracts at 10 to 100 mg mL-1 concentrations in raw 264.7 cells and observed 80% cell viability. The anti-inflammatory effect of B. tinctoria root extract in lipopolysaccharide (LPS)-stimulated Raw 264.7 cells were observed with concentrations at 10, 30, and 50 μg mL-1. The results showed that 77.27-66.82% of nitric oxide (NO) production was inhibited by 50 μg mL-1 B. tinctoria root extract. The protein expression of Inducible NO synthase (iNOS) expression dramatically decreased by 93.14% and 52.65% in raw 264.7 cells treated with water and ethanol extracts of B. tinctoria root, respectively. Moreover, cyclooxygenase-2 (COX-2) protein expression decreased by 42.85% and 69.70% in raw 264.7 cells treated with water and ethanol extracts of B. tinctoria root, respectively. Furthermore, the mRNA expression of pro-inflammatory markers, such as tumor necrosis factor-alpha, interleukin-1β, interleukin-6, monocyte chemoattractant protein-1, and prostaglandin E synthase 2, was significantly suppressed in a concentration-dependent manner. Additionally, the B. tinctoria root extracts effectively inhibited enzymes involved in physiological activities. The B. tinctoria root extracts showed excellent anti-inflammatory effects and can be used as a functional material for biological activities.

Keywords: cytokine, macrophage, pro-inflammatory, protein expression, real-time PCR

Procedia PDF Downloads 71

Authors: Auwal Abubakar, Ahmed Ahidjo, Shazril Imran Shaukat, Noor Khairiah A. Karim, Gokula Kumar Appalanaido, Hafiz Mohd Zin

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Background: The use of optical surface imaging systems (OSISs) is increasingly becoming popular in radiotherapy practice, especially during breast cancer treatment. This study reviews the accuracy of the available commercial OSISs for breast radiotherapy. Method: A literature search was conducted and identified the available commercial OSISs from different manufacturers that are integrated into radiotherapy practice for setup verification during breast radiotherapy. Studies that evaluated the accuracy of the OSISs during breast radiotherapy using cone beam computed tomography (CBCT) as a reference were retrieved and analyzed. The physics and working principles of the systems from each manufacturer were discussed together with their respective strength and limitations. Results: A total of five (5) different commercially available OSISs from four (4) manufacturers were identified, each with a different working principle. Six (6) studies were found to evaluate the accuracy of the systems during breast radiotherapy in conjunction with CBCT as a goal standard. The studies revealed that the accuracy of the system in terms of mean difference ranges from 0.1 to 2.1 mm. The correlation between CBCT and OSIS ranges between 0.4 and 0.9. The limit of agreements obtained using bland Altman analysis in the studies was also within an acceptable range. Conclusion: The OSISs have an acceptable level of accuracy and could be used safely during breast radiotherapy. The systems are non-invasive, ionizing radiation-free, and provide real-time imaging of the target surface at no extra concomitant imaging dose. However, the system should only be used to complement rather than replace x-ray-based image guidance techniques such as CBCT.

Keywords: optical surface imaging system, Cone beam computed tomography (CBCT), surface guided radiotherapy, Breast radiotherapy

Procedia PDF Downloads 66

Authors: Chris Cadman, Marcel Strauss

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Background: In 2020, the University Hospital Wishaw in the United Kingdom became the centre for trauma and orthopaedics within its health board. This resulted in the majority of patients with suspected cauda equina syndrome (CES) being assessed and imaged at this site, putting an increased demand on MR imaging and displacing other previous activity. Following this transition, imaging requests for CES did not always follow national guidelines and would often be missing important clinical and safety information. There also appeared to be a very low positive scan rate compared with previously reported studies. In an attempt to improve patient selection and reduce the burden of CES imaging at this site clinical audit was performed. Methods: A total of 250 consecutive patients imaged to assess for CES were evaluated. Patients had to have presented to either the emergency or orthopaedic department acutely with a presenting complaint of suspected CES. Patients were excluded if they were not admitted acutely or were assessed by other clinical specialities. In total, 233 patients were included. Requests were assessed for appropriate clinical history, accurate and complete clinical assessment and MRI safety information. Clinical assessment was allocated a score of 1-6 based on information relating to history of pain, level of pain, dermatomes/myotomes affected, peri-anal paraesthesia/anaesthesia, anal tone and post-void bladder volume with each element scoring one point. Images were assessed for positive findings of CES, acquired spinal stenosis or nerve root compression. Results: Overall, 73% of requests had a clear clinical history of CES. The urgency of the request for imaging was given in 23% of cases. The mean clinical assessment score was 3.7 out of a total of 6. Overall, 2% of scans were positive for CES, 29% had acquired spinal stenosis and 30% had nerve root compression. For patients with CES, 75% had acute neurological signs compared with 68% of the study population. CES patients had a mean clinical history score of 5.3 compared with 3.7 for the study population. Overall, 95% of requests had appropriate MRI safety information. Discussion: it study included 233 patients who underwent specialist assessment and referral for MR imaging for suspected CES. Despite the serious nature of this condition, a large proportion of imaging requests did not have a clear clinical query of CES and the level of urgency was not given, which could potentially lead to a delay in imaging and treatment. Clinical examination was often also incomplete, which can make triaging of patients presenting with similar symptoms challenging. The positive rate for CES was only 2%, much below other studies which had positive rates of 6–40% with a large meta-analysis finding a mean positive rate of 19%. These findings demonstrate an opportunity to improve the quality of imaging requests for suspected CES. This may help to improve patient selection for imaging and result in a positive rate for CES imaging that is more in line with other centres.

Keywords: cauda equina syndrome, acute back pain, MRI, spine

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Authors: M. G. R. S. Perera, B. S. Weerakoon, L. P. G. Sherminie, M. L. Jayatilake, R. D. Jayasinghe, W. Huang

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The aim of this study was to measure and evaluate the Longitudinal Relaxation Times (T1) in bone marrow of an Acute Myeloid Leukaemia (AML) patient in order to explore the potential for a prognostic biomarker using Magnetic Resonance Imaging (MRI) which will be a non-invasive prognostic approach to AML. MR image data were collected in the DICOM format and MATLAB Simulink software was used in the image processing and data analysis. For quantitative MRI data analysis, Region of Interests (ROI) on multiple image slices were drawn encompassing vertebral bodies of L3, L4, and L5. T1 was evaluated using the T1 maps obtained. The estimated bone marrow mean value of T1 was 790.1 (ms) at 3T. However, the reported T1 value of healthy subjects is significantly (946.0 ms) higher than the present finding. This suggests that the T1 for bone marrow can be considered as a potential prognostic biomarker for AML patients.

Keywords: acute myeloid leukaemia, longitudinal relaxation time, magnetic resonance imaging, prognostic biomarker.

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Authors: Vallisree Sivathanu, Kumaraswamidhas Lakshmi Annamalai, Trupti Ranjan Lenka

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We report a tandem solar cell model with Silicon as the bottom cell absorber material and Cu₂BaSn(S, Se)₄(CBTSSe) as absorber material for the top cell. As a first step, the top and bottom cells were modelled and validated by comparison with the experiment. Once the individual cells are validated, then the tandem structure is modelled with Indium Tin Oxide(ITO) as conducting layer between the top and bottom cells. The tandem structure yielded better open circuit voltage and fill factor; however, the efficiency obtained is 7.01%. The top cell and the bottom cells are investigated with the help of electron-hole current density, photogeneration rate, and external quantum efficiency profiles. In order to minimize the various loss mechanisms in the tandem solar cell, the material parameters are optimized within experimentally achievable limits. Initially, the top cell optimization was carried out; then, the bottom cell is optimized for maximizing the light absorption, and upon minimizing the current and photon losses in the tandem structure, the maximum achievable efficiency is predicted to be 19.52%.

Keywords: CBTSSe, silicon, tandem, solar cell, device modeling, current losses, photon losses

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Authors: Jameela Ali, Hamid A. Jalab, Loay E. George, Abdul Rahim Ahmad, Azizah Suliman, Karim Al-Jashamy

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Red blood cells (RBC) are the most common types of blood cells and are the most intensively studied in cell biology. The lack of RBCs is a condition in which the amount of hemoglobin level is lower than normal and is referred to as “anemia”. Abnormalities in RBCs will affect the exchange of oxygen. This paper presents a comparative study for various techniques for classifying the RBCs as normal, or abnormal (anemic) using WEKA. WEKA is an open source consists of different machine learning algorithms for data mining applications. The algorithm tested are Radial Basis Function neural network, Support vector machine, and K-Nearest Neighbors algorithm. Two sets of combined features were utilized for classification of blood cells images. The first set, exclusively consist of geometrical features, was used to identify whether the tested blood cell has a spherical shape or non-spherical cells. While the second set, consist mainly of textural features was used to recognize the types of the spherical cells. We have provided an evaluation based on applying these classification methods to our RBCs image dataset which were obtained from Serdang Hospital-alaysia, and measuring the accuracy of test results. The best achieved classification rates are 97%, 98%, and 79% for Support vector machines, Radial Basis Function neural network, and K-Nearest Neighbors algorithm respectively.

Keywords: K-nearest neighbors algorithm, radial basis function neural network, red blood cells, support vector machine

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Authors: Sameer Masood, Lucas Chartier

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Head injuries are a commonly encountered presentation in emergency departments (ED) and the Choosing Wisely Canada (CWC) campaign was released in June 2015 in an attempt to decrease imaging utilization for patients with minor head injuries. The impact of the CWC campaign on imaging utilization for head injuries has not been explored in the ED setting. In our study, we describe the characteristics of patients with head injuries presenting to a tertiary care academic ED and the impact of the CWC campaign on CT head utilization. This retrospective cohort study used linked databases from the province of Ontario, Canada to assess emergency department visits with a primary diagnosis of head injury made between June 1, 2014 and Aug 31, 2016 at the University Health Network in Toronto, Canada. We examined the number of visits during the study period, the proportion of patients that had a CT head performed before and after the release of the CWC campaign, as well as mode of arrival, and disposition. There were 4,322 qualifying visits at our site during the study period. The median presenting age was 44.12 years (IQR 27.83,67.45), the median GCS was 15 (IQR 15,15) and the majority of patients presenting had intermediate acuity (CTAS 3). Overall, 43.17% of patients arrived via ambulance, 49.24 % of patients received a CT head and 10.46% of patients were admitted. Compared to patients presenting before the CWC campaign release, there was no significant difference in the rate of CT heads after the CWC (50.41% vs 47.68%, P = 0.07). There were also no significant differences between the two groups in mode of arrival (ambulance vs ambulatory) (42.94% vs 43.48%, P = 0.72) or admission rates (9.85% vs 11.26%, P = 0.15). However, more patients belonged to the high acuity groups (CTAS 1 or 2) in the post CWC campaign release group (12.98% vs 8.11% P <0.001). Visits for head injuries make up a significant proportion of total ED visits and approximately half of these patients receive CT imaging in the ED. The CWC campaign did not seem to impact imaging utilization for head injuries in the 14 months following its launch. Further efforts, including local quality improvement initiatives, are likely needed to increase adherence to its recommendation and reduce imaging utilization for head injuries.

Keywords: choosing wisely, emergency department, head injury, quality improvement

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Authors: Mohamed A. Dkhil, Denis Delic, Saleh Al-Quraishy

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Coccidiosis causes considerable economic loss in the poultry industry. The current study aimed to investigate the response of goblet cells as well as the induced tissue damage during Eimeria papilata infection. Mice were infected with sporulated E. papillata oocyts. On day 5 post-infection, the fecal output was determined. Also, the jejunum was prepared for the histological, histochemical and molecular studies. Our results revealed that the intestinal coccidian infection with E. papillata induced a marked goblet cell hypoplasia and depleted mucus secretion. Also, the infection was able to alter the jejuna architecture and increased the apoptotic cells inside the villi. In addition, the real time PCR results indicated that, the inflammatory cytokines TNF-α, iNOS, IFN-y and IL-1β were significantly up-regulated. In contrast, the mRNA expression patterns of IL-6 in response to E. papillata infection did not differ significantly between control and infected mice. Moreover, the mRNA expression of TLR4 was significantly up-regulated, whereas the expression of MUC2 is significantly down-regulated upon infection. Further studies are required to understand the regulatory mechanisms of goblet cells related genes.

Keywords: goblet cells, Eimeria papillata, mice, jejunum

Procedia PDF Downloads 277

Authors: Misa Nakao, Yuta Kurashina, Chikahiro Imashiro, Kenjiro Takemura

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The Research Goal: With the growing demand for regenerative medicine, an effective mass cell culture process is required. In a repetitive subculture process for proliferating cells, preparing single cell suspension which does not contain any cell aggregates is highly required because cell aggregates often raise various undesirable phenomena, e.g., apoptosis and decrease of cell proliferation. Since cell aggregates often occur in cell suspension during conventional subculture processes, this study proposes a single cell sorter driven by a resonance vibration of a cell culture substrate. The Method and the Result: The single cell sorter is simply composed of a cell culture substrate and a glass pipe vertically placed against the cell culture substrate with a certain gap corresponding to a cell diameter. The cell culture substrate is made of biocompatible stainless steel with a piezoelectric ceramic disk glued to the bottom side. Applying AC voltage to the piezoelectric ceramic disk, an out-of-plane resonance vibration with a single nodal circle of the cell culture substrate can be excited at 5.5 kHz. By doing so, acoustic radiation force is emitted, and then cell suspension containing only single cells is pumped into the pipe and collected. This single cell sorter is effective to collect single cells selectively in spite of its quite simple structure. We collected C2C12 myoblast cell suspension by the single cell sorter with the vibration amplitude of 12 µmp-p and evaluated the ratio of single cells in number against the entire cells in the suspension. Additionally, we cultured the collected cells for 72 hrs and measured the number of cells after the cultivation in order to evaluate their proliferation. As a control sample, we also collected cell suspension by conventional pipetting, and evaluated the ratio of single cells and the number of cells after the 72-hour cultivation. The ratio of single cells in the cell suspension collected by the single cell sorter was 98.2%. This ratio was 9.6% higher than that collected by conventional pipetting (statistically significant). Moreover, the number of cells cultured for 72 hrs after the collection by the single cell sorter yielded statistically more cells than that collected by pipetting, resulting in a 13.6% increase in proliferated cells. These results suggest that the cell suspension collected by the single cell sorter driven by the resonance vibration hardly contains cell aggregates whose diameter is larger than the gap between the cell culture substrate and the pipe. Consequently, the cell suspension collected by the single cell sorter maintains high cell proliferation. Conclusions: In this study, we developed a single cell sorter capable of sorting and pumping single cells by a resonance vibration of a cell culture substrate. The experimental results show the single cell sorter collects single cell suspension which hardly contains cell aggregates. Furthermore, the collected cells show higher proliferation than that of cells collected by conventional pipetting. This means the resonance vibration of the cell culture substrate can benefit us with the increase in efficiency of mass cell culture process for clinical applications.

Keywords: acoustic radiation force, cell proliferation, regenerative medicine, resonance vibration, single cell sorter

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Authors: Jameela Ali Alkrimi, Abdul Rahim Ahmad, Azizah Suliman, Loay E. George

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Red blood cells (RBCs) are among the most commonly and intensively studied type of blood cells in cell biology. The lack of RBCs is a condition characterized by lower than normal hemoglobin level; this condition is referred to as 'anemia'. In this study, a software was developed to isolate RBCs by using a machine learning approach to classify anemic RBCs in microscopic images. Several features of RBCs were extracted using image processing algorithms, including principal component analysis (PCA). With the proposed method, RBCs were isolated in 34 second from an image containing 18 to 27 cells. We also proposed that PCA could be performed to increase the speed and efficiency of classification. Our classifier algorithm yielded accuracy rates of 100%, 99.99%, and 96.50% for K-nearest neighbor (K-NN) algorithm, support vector machine (SVM), and neural network ANN, respectively. Classification was evaluated in highly sensitivity, specificity, and kappa statistical parameters. In conclusion, the classification results were obtained for a short time period with more efficient when PCA was used.

Keywords: red blood cells, pre-processing image algorithms, classification algorithms, principal component analysis PCA, confusion matrix, kappa statistical parameters, ROC

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Authors: Faris M. Altaf, Abdullah M Elkeshy

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Forty-two albino male rats aged between 30 and 40 days (weighted 200 g to 250 g) were used in the present study. The left sural nerves of 36 rats were subjected to crush injury at 1 to 6 weeks intervals using 6 animals at each interval. The right and left sural nerves of the rest 6 rats were used as a control. After 2 weeks of the crush injury, the endoneurium showed channel-like spaces that were lined by the fibroblast-like cells and collagen bundles. These channels contained degenerated myelin and were connected with the perivascular and subperineurial spaces. Some of the flattened fibroblast-like cells were arranged in several layers in the subperineurial and perivascular spaces, forming barrier-like cellular sheets localizing the endoneurial edema in these spaces. Fibroblast-like cells also wrapped the regenerating nerve fibers by their branching cytoplasmic processes. At the end of the third week, the flattened fibroblasts formed nearly continuous sheets in the subperineurial and perivascular spaces. Macrophages were frequently noticed between these cellular barrier-like sheets and in the subperineurial and perivascular spaces. Conclusion: it could be concluded that the endoneurial fibroblast-like cells form barrier-like cellular sheets that localized the endoneurial edema in the subperineurial and perivascular spaces and create also the endoneurial channel-like spaces containing degenerated myelin and endoneurial edema helping the resolution of such edema.

Keywords: sural nerve, endoneurial fibroblast-like cells, endoneurial edema, barrier-like and channel-like spaces

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Authors: Sung-Hyeon Park, Jin-Wook Jang, Hak-Sung Kim

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In recent years, Terahertz (THz) time-domain spectroscopy (TDS) imaging method has been received considerable interest as a promising non-destructive technique for detection of internal defects. In comparison to other non-destructive techniques such as x-ray inspection method, scanning acoustic tomograph (SAT) and microwave inspection method, THz-TDS imaging method has many advantages: First, it can measure the exact thickness and location of defects. Second, it doesn’t require the liquid couplant while it is very crucial to deliver that power of ultrasonic wave in SAT method. Third, it didn’t damage to materials and be harmful to human bodies while x-ray inspection method does. Finally, it exhibits better spatial resolution than microwave inspection method. However, this technology couldn’t be applied to IC package because THz radiation can penetrate through a wide variety of materials including polymers and ceramics except of metals. Therefore, it is difficult to detect the defects in IC package which are composed of not only epoxy and semiconductor materials but also various metals such as copper, aluminum and gold. In this work, we proposed a special method for detecting the void in the IC package using THz-TDS imaging system. The IC package specimens for this study are prepared by Packaging Engineering Team in Samsung Electronics. Our THz-TDS imaging system has a special reflection mode called pitch-catch mode which can change the incidence angle in the reflection mode from 10 o to 70 o while the others have transmission and the normal reflection mode or the reflection mode fixed at certain angle. Therefore, to find the voids in the IC package, we investigated the appropriate angle as changing the incidence angle of THz wave emitter and detector. As the results, the voids in the IC packages were successfully detected using our THz-TDS imaging system.

Keywords: terahertz, non-destructive technique, void, IC package

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Authors: Mahdi Farajzadeh Ajirlou

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Ordinary fetal brain development can be considered by in vivo attractive reverberation imaging (MRI) from the 18th gestational week (GW) to term and depends fundamentally on T2-weighted and diffusion-weighted (DW) arrangements. The foremost commonly suspected brain pathologies alluded to fetal MRI for assist assessment are ventriculomegaly, lost corpus callosum, and anomalies of the posterior fossa. Brain division could be a crucial to begin with step in neuroimage examination. Within the case of fetal MRI it is especially challenging and critical due to the subjective introduction of the hatchling, organs that encompass the fetal head, and irregular fetal movement. A few promising strategies have been proposed but are constrained in their execution in challenging cases and in realtime division. Fetal MRI is routinely performed on a 1.5-Tesla scanner without maternal or fetal sedation. The mother lies recumbent amid the course of the examination, the length of which is ordinarily 45 to 60 minutes. The accessibility and continuous approval of standardizing fetal brain development directions will give critical devices for early discovery of impeded fetal brain development upon which to oversee high-risk pregnancies.

Keywords: brain, fetal, MRI, imaging

Procedia PDF Downloads 79

Authors: Prabesh Karki, Shyam Thapa Chettri, Bajarang Prasad Sah, Manoj Bhattarai, Sudeep Mishra

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Introduction: Frontal sinus is frequently described as the most difficult sinus to access surgically due to its proximity to the cribriform plate, orbit, and anterior ethmoid artery. Frontal sinus surgery requires a detailed understanding of the cellular structure and FSDP unique to each patient, making high-resolution CT scans an indispensable tool to assess the difficulty of planned sinus surgery. International Frontal Sinus Anatomy Classification (IFAC) was developed to provide a more precise nomenclature for cells in the frontal recess, classifying cells based on their anatomic origin. Objectives: To assess the proportion of frontal cell variants defined by IFAC, variation with respect to age and gender. Methods: 54 cases were enrolled after a detailed clinical history, thorough general and physical examinations, and CT a report ordered in a film. Assessment and tabulation of the presence of frontal cells according to the IFAC analyzed. The prevalence of each cell type was calculated, and data were entered in MS Excel and analyzed using Statistical Package for the Social Sciences (SPSS). Descriptive statistics and frequencies were defined for categorical and numerical variables. Frequency, percentage, the mean and standard deviation were calculated. Result: Among 54 patients, 30 (55.6%) were male and 24 (44.4%) were female. The patient enrolled ranged from 18 to 78 years. Majority33.3% (n=18) were in age group of >50 years.According to IFAC, Agger nasi cells (92.6%) were most common, whereas supraorbital ethmoidal cells were least common 16 (29.6%). Prevalence of other frontoethmoidal cells was SAC- 57.4%, SAFC- 38.9%, SBC- 74.1%, SBFC- 33.3%, FSC- 38.9% of 54 cases. Conclusion: IFAC is an international consensus document that describes an anatomically precise nomenclature for classifying frontoethmoidal cells' anatomy. This study has defined the prevalence, symmetry and reliability of frontoethmoidal cells as established by the IFAC system as in other parts of the world.

Keywords: frontal sinus, frontoethmoidal cells, international frontal sinus anatomy classification

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Authors: Ahmed Gaber Abdel Raheem, Nashwa Ahmed Mohamed

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Introduction: Sensorineural hearing loss (SNHL) is largely caused by the degeneration of the cochlea. Therapeutic options for SNHL are limited to hearing aids and cochlear implants. The cell transplantation approach to the regeneration of hair cells has gained considerable attention because stem cells are believed to accumulate in the damaged sites and have the potential for the repair of damaged tissues. The aim of the work: was to assess the use of bone marrow transplantation in repair of damaged inner ear hair cells in rats after the damage had been inflicted by Amikacin injection. Material and Methods: Thirty albino rats were used in this study. They were divided into three groups. Each group ten rats. Group I: used as control. Group II: Were given Amikacin- intratympanic injection till complete loss of hearing function. This could be assessed by Distortion product Otoacoustic Emission (DPOAEs) and / or auditory brain stem evoked potential (ABR). GroupIII: were given intra-peritoneal injection of bone marrow stem cell after complete loss of hearing caused by Amikacin. Clinical assessment was done using DPOAEs and / or auditory brain stem evoked potential (ABR), before and after bone marrow injection. Histological assessment of the inner ear was done by light and electron microscope. Also, Detection of stem cells in the inner ear by immunohistochemistry. Results: Histological examination of the specimens showed promising improvement in the structure of cochlea that may be responsible for the improvement of hearing function in rats detected by DPOAEs and / or ABR. Conclusion: Bone marrow stem cells transplantation might be useful for the treatment of SNHL.

Keywords: amikacin, hair cells, sensorineural hearing loss, stem cells

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Authors: Phuriwat Laomethakorn, Siritron Samosorn, Ramida Watanapokasin

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Cancer metastasis is the major cause of cancer-related death. Therefore searching for a compound that could inhibit cancer metastasis is necessary. 13-Butoxyberberine bromide is a berberine derivative that has not been reported an anti-metastatic effect on skin cancer cells. This study aimed to investigate the anti-metastatic effect of 13-butoxyberberine bromide on skin cancer A431 cells. The effect of 13-butoxyberberine bromide on A431 cell viability was examined by MTT assay. Suppression of cell migration and invasion in A431 cells were determined by wound healing assay, transwell migration assay, and transwell invasion assay. Metastasis proteins were determined by western blotting. The results demonstrated that 13-butoxyberberine bromide decreased A431 cell viability in a dose-dependent manner. In addition, sub-toxic concentrations of 13-butoxyberberine bromide suppressed cell migration and invasion in A431 cells. In addition, 13-butoxyberberine bromide showed anti-metastatic effects by down-regulated MMP-2 and MMP-9 expression. These findings may be useful in the development of 13-butoxyberberine bromide as an anti-metastatic drug in the future.

Keywords: 13-butoxyberberine bromide, metastasis, skin cancer, MMP

Procedia PDF Downloads 104

Authors: Ruth Goldschmidt, Vyacheslav Kalchenko, Lilah Agemy, Rachel Elmoalem, Avigdor Scherz

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Vascular Targeted Photodynamic therapy (VTP) is a new modality for selective cancer treatment that leads to the complete tumor ablation. A photosensitizer, a bacteriochlorophyll derivative in our case, is first administered to the patient and followed by the illumination of the tumor area, by a near-IR laser for its photoactivation. The photoactivated drug releases reactive oxygen species (ROS) in the circulation, which reacts with blood cells and the endothelium leading to the occlusion of the blood vasculature. If the blood vessels are only partially closed, the tumor may recover, and cancer cells could survive. On the other hand, excessive treatment may lead to toxicity of healthy tissues nearby. Simultaneous VTP monitoring and image processing independent of the photoexcitation laser has not yet been reported, to our knowledge. Here we present a method for blood flow monitoring, using a real-time laser speckle imaging (RTLSI) in the tumor during VTP. We have synthesized over the years a library of bacteriochlorophyll derivatives, among them WST11 and STL-6014. Both are water soluble derivatives that are retained in the blood vasculature through their partial binding to HSA. WST11 has been approved in Mexico for VTP treatment of prostate cancer at a certain drug dose, and time/intensity of illumination. Application to other bacteriochlorophyll derivatives or other cancers may require different treatment parameters (such as light/drug administration). VTP parameters for STL-6014 are still under study. This new derivative mainly differs from WST11 by its lack of the central Palladium, and its conjugation to an Arg-Gly-Asp (RGD) sequence. RGD is a tumor-specific ligand that is used for targeting the necrotic tumor domains through its affinity to αVβ3 integrin receptors. This enables the study of cell-targeted VTP. We developed a special RTLSI module, based on Labview software environment for data processing. The new module enables to acquire raw laser speckle images and calculate the values of the laser temporal statistics of time-integrated speckles in real time, without additional off-line processing. Using RTLSI, we could monitor the tumor’s blood flow following VTP in a CT26 colon carcinoma ear model. VTP with WST11 induced an immediate slow down of the blood flow within the tumor and a complete final flow arrest, after some sporadic reperfusions. If the irradiation continued further, the blood flow stopped also in the blood vessels of the surrounding healthy tissue. This emphasizes the significance of light dose control. Using our RTLSI system, we could prevent any additional healthy tissue damage by controlling the illumination time and restrict blood flow arrest within the tumor only. In addition, we found that VTP with STL-6014 was the most effective when the photoactivation was conducted 4h post-injection, in terms of tumor ablation success in-vivo and blood vessel flow arrest. In conclusion, RTSLI application should allow to optimize VTP efficacy vs. toxicity in both the preclinical and clinical arenas.

Keywords: blood vessel occlusion, cancer treatment, photodynamic therapy, real time imaging

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Authors: Laura Gantley, Vanessa M. Conn, Stuart Webb, Kirsty Kirk, Marta Gabryelska, Duncan Holds, Brett W. Stringer, Simon J. Conn

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Trinucleotide repeat disorders comprise ~20 severe, inherited human neuromuscular and neurodegenerative disorders, which are a result of an abnormal expansion of repetitive sequences in the DNA. The most common of these, Huntington’s disease, results from the expansion of the CAG repeat region in exon 1 of the HTT gene via an unknown mechanism. Non-coding RNAs have been implicated in the initiation and progression of many diseases; thus, we focus on one circular RNA (circRNA) molecule arising from non-canonical splicing (back splicing) of HTT pre-mRNA. This circRNA and its mouse orthologue were transgenically overexpressed in human cells (SHSY-5Y and HEK293T) and mouse cells (Mb1), respectively. High-content imaging and flow cytometry demonstrated the overexpression of this circRNA reduces cell proliferation, reduces nuclear size independent of cellular size, and alters cell cycle progression. Analysis of protein by western blot and immunofluorescence demonstrated no change to HTT protein levels but altered nuclear-cytoplasmic distribution without impacting the expansion of the HTT repeat region. As these phenotypic and genotypic changes are found in Huntington’s disease patients, these results may suggest that this circRNA may play a functional role in the progression of Huntington’s disease.

Keywords: cell biology, circular RNAs, Huntington’s disease, molecular biology, neurodegenerative disorders

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Authors: Sana Abbasa, Shahzad Bhattiab, Nadir Khan

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Sargassum siliquastrum is brown seaweed with traditional claims for some medicinal properties. This research was done to investigate the methanol extract of S. siliquastrum for antiproliferative activity against human cervical cancer cell line, HeLa and its mode of cell death. From methylene blue assay, S. siliquastrum exhibited antiproliferative activity on HeLa cells with IC50 of 3.87 µg/ml without affecting non-malignant cells. Phase contrast microscopy indicated the confluency reduction in HeLa cells and changes on the cell shape. Nuclear staining with Hoechst 33258 displayed the formation of apoptotic bodies and fragmented nuclei. S. siliquastrum also induced early apoptosis event in HeLa cells as confirmed by FITC-Annexin V/propidium iodide staining by flow cytometry analysis. Cell cycle analysis indicated growth arrest of HeLa cells at G1/S phase. Protein study by flow cytometry indicated the increment of p53, slight increase of Bax and unchanged level of Bcl-2. In conclusion, S. siliquastrum demonstrated an antiproliferative activity in HeLa cell by inducing G1/S cell cycle arrest via p53-mediated pathway.

Keywords: sargassum siliquastrum, cervical cancer, P53, antiproleferation

Procedia PDF Downloads 633

Authors: Ankita Singh, Chandan Gorain, Amirul I. Mallick

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Successful adherence of the mucosal epithelial cells is the key early step for Campylobacter jejuni pathogenesis (C. jejuni). A set of Surface Exposed Colonization Proteins (SECPs) are among the major factors involved in host cell adherence and invasion of C. jejuni. Among them, constitutively expressed surface-exposed lipoprotein adhesin of C. jejuni, JlpA, interacts with intestinal heat shock protein 90 (hsp90α) and contributes in disease progression by triggering pro-inflammatory response via activation of NF-κB and p38 MAP kinase pathway. Together with its ability to express in the bacterial surface, higher sequence conservation and predicted predominance of several B cells epitopes, JlpA protein reserves its potential to become an effective vaccine candidate against wide range of Campylobacter sps including C. jejuni. Given that chickens are the primary sources for C. jejuni and persistent gut colonization remain as major cause for foodborne pathogenesis to humans, present study explicitly used chickens as model to test the immune-protective efficacy of JlpA protein. Taking into account that gastrointestinal tract is the focal site for C. jejuni colonization, to extrapolate the benefit of mucosal (intragastric) delivery of JlpA protein, a food grade Nisin inducible Lactic acid producing bacteria, Lactococcus lactis (L. lactis) was engineered to express recombinant JlpA protein (rJlpA) in the surface of the bacteria. Following evaluation of optimal surface expression and functionality of recombinant JlpA protein expressed by recombinant L. lactis (rL. lactis), the immune-protective role of intragastric administration of live rL. lactis was assessed in commercial broiler chickens. In addition to the significant elevation of antigen specific mucosal immune responses in the intestine of chickens that received three doses of rL. lactis, marked upregulation of Toll-like receptor 2 (TLR2) gene expression in association with mixed pro-inflammatory responses (both Th1 and Th17 type) was observed. Furthermore, intragastric delivery of rJlpA expressed by rL. lactis, but not the injectable form, resulted in a significant reduction in C. jejuni colonization in chickens suggesting that mucosal delivery of live rL. lactis expressing JlpA serves as a promising vaccine platform to induce strong immune-protective responses against C. jejuni in chickens.

Keywords: chickens, lipoprotein adhesion of Campylobacter jejuni, immuno-protection, Lactococcus lactis, mucosal delivery

Procedia PDF Downloads 140

Authors: A. Apostolopoulou, D. Sygkridou, A. N. Kalarakis, E. Stathatos

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Mesoscopic perovskite solar cells (mp-PSCs) with mesoporous bilayer were fabricated under ambient conditions. The bilayer was formed by capping the mesoporous TiO₂ layer with a layer of In₂O₃. CH₃NH₃I_3-xCl_x mixed halide perovskite was prepared through the one-step method and was used as the light absorber. The mp-PSCs with the composite TiO₂/In₂O₃ mesoporous layer exhibited optimized electrical parameters, compared with the PSCs that employed only a TiO₂ mesoporous layer, with a current density of 23.86 mA/cm², open circuit voltage of 0.863 V, fill factor of 0.6 and a power conversion efficiency of 11.2%. These results indicate that the formation of a proper semiconductor capping layer over the basic TiO₂ mesoporous layer can facilitate the electron transfer, suppress the recombination and subsequently lead to higher charge collection efficiency.

Keywords: ambient conditions, high efficiency solar cells, mesoscopic perovskite solar cells, TiO₂ / In₂O₃ bilayer

Procedia PDF Downloads 271