Search results for: live cells imaging

Authors: Nitin Dwivedi, Jigna Shah

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Brain tumor is metastatic neoplasm of central nervous system, in most of cases it is life threatening disease with low survival rate. Despite of enormous efforts in the development of therapeutics and diagnostic tools, the treatment of brain tumors and gliomas remain a considerable challenge in the area of neuro-oncology. The most reason behind of this the presence of physiological barriers including blood brain barrier and blood brain tumor barrier, lead to insufficient reach ability of therapeutic agents at the site of tumor, result of inadequate destruction of gliomas. So there is an indeed need empowerment of brain tumor imaging for better characterization and delineation of tumors, visualization of malignant tissue during surgery, and tracking of response to chemotherapy and radiotherapy. Multifunctional different generations of dendrimer offer an improved effort for potentiate drug delivery at the site of brain tumor and gliomas. So this article emphasizes the innovative dendrimer approaches in tumor targeting, tumor imaging and delivery of therapeutic agent.

Keywords: blood brain barrier, dendrimer, gliomas, nanotechnology

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Authors: Sri Ratna Rahayu, Oktia Woro Kasmini Handayani, Lourensiana Y. S. Ngaga, Imade Sudana, Irwan Budiono

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Malaria is an infectious disease that still cannot be solved in Kori village, West Nusa Tenggara, Indonesia, where the most of people live in rumah panggung (Stilts House). The purpose of this study was to know whether there were the effects of rumah panggung environment, social culture, and behavior on malaria incidence in the Kori village. A cross-sectional study was performed to explore the effects of rumah panggung environment, social culture and behavior on malaria incidence. This study recruited 280 respondents, who live in the rumah panggung, permanent residents in Kori village, were age above 17 years old, and suffered from malaria in the past year. The collected data were analyzed with path analysis. The results of this study showed that the environment of rumah panggung and behavior have a direct effect on the incidence of malaria (p < 0.05). It could be concluded that improvement of environmental conditions of rumah panggung, sociocultural, and behavioral changes to maintain a healthy environment are needed to reduce the malaria incidence.

Keywords: Rumah panggung, socio-cultural, behavior, Malaria

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Authors: Muhammad Younus, Muhammad Sajid, Muti-ur-Rehman Khan, Aftab Ahmad Anjum, Muhammad Asif Idrees, Iahtasham Khan, Aman Ullah Khan, Sajid Umar, Raheela Akhtar

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The present study was conducted to investigate the hazardous effects of lead on health and edible organs of Lohi sheep. The adult Lohi sheep (n=48) were randomly divided into two equal groups. The first group was administered lead acetate at dose of 70 mg/kg live body weight daily as 10% solution by oral route for a period of 90 days and the second group served as a negative control. Blood and tissue samples were collected at day 0, 30, 60 and 90 and analyzed for lead concentration by atomic absorption spectrophotometry. The kidney showed the highest lead concentration (p < 0.05) followed by liver and then muscle. Lead acetate treated sheep showed structural and behavioral changes during the last month of trial. Liver showed necrosis, hemorrhages and hyperactivation of macrophages. Kidney showed degenerative and necrotic changes in glomeruli and tubules and the characteristic intranuclear inclusion bodies in tubular epithelial cells on H and E staining. It was concluded that Lohi sheep is affected by lead intoxication at low dose for longer period and hence exhibits lead accumulation in edible tissues.

Keywords: Lohi sheep, lead acetate, edible tissue, histopathology

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Authors: K. Żak, S. Przetocka, R. Kitel, K. Guzik, B. Musielak, S. Malicki, G. Dubin, T. A. Holak

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Studies on tumor genesis revealed a number of factors that may potentially serve as molecular targets for immunotherapies. One of such promising targets are PD1 and PDL1 proteins. PD1 (Programmed cell death protein 1) is expressed by activated T cells and plays a critical role in modulation of the host's immune response. One of the PD1 ligands -PDL1- is expressed by macrophages, monocytes and cancer cells which exploit it to avoid immune attack. The notion of the mechanisms used by cancer cells to block the immune system response was utilized in the development of therapies blocking PD1-PDL1 interaction. Up to date, human PD1-PDL1 complex has not been crystallized and structure of the mouse-human complex does not provide a complete view of the molecular basis of PD1-PDL1 interactions. The purpose of this study is to obtain crystal structure of the human PD1-PDL1 complex which shall allow rational design of small molecule inhibitors of the interaction. In addition, the study presents results of binding small-molecules to PD1 and fragment docking towards PD1 protein which will facilitate the design and development of small–molecule inhibitors of PD1-PDL1 interaction.

Keywords: PD1, PDL1, cancer, small molecule, drug discovery

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Authors: Qing Cao, Fei Wang, Yu-Qiang Wang, Li-Ya Huang, Tian-Tian Sang, Shu-Yan Chen

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Aims: TNF Receptor-Associated Factor 6 (TRAF6) acts as a multifunctional regulator of the Transforming Growth Factor (TGF)-β signaling pathway, and mediates Smad-independent JNK and p38 activation via TGF-β. This study was performed to test the hypothesis that TGF-β/TRAF6 is essential for angiotensin-II (Ang II)-induced differentiation of rat c-kit+ Cardiac Stem Cells (CSCs). Methods and Results: c-kit+ CSCs were isolated from neonatal Sprague Dawley (SD) rats, and their c-kit status was confirmed with immunofluorescence staining. A TGF-β type I receptor inhibitor (SB431542) or the small interfering RNA (siRNA)-mediated knockdown of TRAF6 were used to investigate the role of TRAF6 in TGF-β signaling. Rescue of TRAF6 siRNA transfected cells with a 3'UTR deleted siRNA insensitive construct was conducted to rule out the off target effects of the siRNA. TRAF6 dominant negative (TRAF6DN) vector was constructed and used to infect c-kit+ CSCs, and western blotting was used to assess the expression of TRAF6, JNK, p38, cardiac-specific proteins, and Wnt signaling proteins. Physical interactions between TRAF6 and TGFβ receptors were studied by coimmunoprecipitation. Cardiac differentiation was suppressed in the absence of TRAF6. Forced expression of TRAF6 enhanced the expression of TGF-β-activated kinase1 (TAK1), and inhibited Wnt signaling. Furthermore, TRAF6 increased the expression of cardiac-specific proteins (cTnT and Cx-43) but inhibited the expression of Wnt3a. Conclusions: Our data suggest that TRAF6 plays an important role in Ang II induced differentiation of c-kit+ CSCs via the non-canonical signaling pathway.

Keywords: cardiac stem cells, differentiation, TGF-β, TRAF6, ubiquitination, Wnt

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Authors: Maryam Parsian, Pelin Mutlu, Ufuk Gunduz

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Two-dimensional cell culture affords simplicity and low cost, but it has serious limitations; lacking cell-cell and cell-matrix interactions that are present in tissues. Cancer cells grown in 3D culture systems have distinct phenotypes of adhesion, growth, migration, invasion as well as profiles of gene and protein expression. These interactions cause the 3D-cultured cells to acquire morphological and cellular characteristics relevant to in vivo tumors. Palbociclib is a chemotherapeutic agent for the treatment of ER-positive and HER-negative metastatic breast cancer. Poly-amidoamine (PAMAM) dendrimer is a well-defined, special three-dimensional structure and has a multivalent surface and internal cavities that can play an essential role in drug delivery systems. In this study, palbociclib is loaded onto the magnetic PAMAM dendrimer. Hanging droplet method was used in order to form 3D spheroids. The possible toxic effects of both free drug and drug loaded nanoparticles were evaluated in 2D and 3D MCF-7, MD-MB-231 and SKBR-3 breast cancer cell culture models by performing MTT cell viability and Alamar Blue assays. MTT analysis was performed with six different doses from 1000 µg/ml to 25 µg/ml. Drug unloaded PAMAM dendrimer did not demonstrate significant toxicity on all breast cancer cell lines. The results showed that 3D spheroids are clearly less sensitive than 2D cell cultures to free palbociclib. Also, palbociclib loaded PAMAM dendrimers showed more toxic effect than free palbociclib in all cell lines at 2D and 3D cultures. The results suggest that the traditional cell culture method (2D) is insufficient for mimicking the actual tumor tissue. The response of the cancer cells to anticancer drugs is different in the 2D and 3D culture conditions. This study showed that breast cancer cells are more resistant to free palbociclib in 3D cultures than in 2D cultures. However, nanoparticle loaded drugs can be more cytotoxic when compared to free drug.

Keywords: 2D and 3D cell culture, breast cancer, palbociclibe, PAMAM magnetic nanoparticles

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Authors: Gehan ElAkabawy

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Background: Diabetes mellitus causes severe deteriorations of almost all the organs and systems of the body, as well as significant damage to the oral cavity. The oral changes are mainly related to salivary glands dysfunction characterized by hyposalivation and xerostomia, which significantly reduce diabetic patients’ quality of life. Human dental pulp stem cells represent a promising source for cell-based therapies, owing to their easy, minimally invasive surgical access, and high proliferative capacity. It was reported that the trophic support mediated by dental pulp stem cells can rescue the functional and structural alterations of damaged salivary glands. However, potential differentiation and paracrine effects of human dental pulp stem cells in diabetic-induced parotid gland damage have not been previously investigated. Our study aimed to investigate the therapeutic effects of intravenous transplantation of human dental pulp stem cells (hDPSCs) on parotid gland injury in a rat model of streptozotocin (STZ)-induced type 1 diabetes. Methods: Thirty Sprague-Dawley male rats were randomly categorised into three groups: control, diabetic (STZ), and transplanted (STZ+hDPSCs). hDPSCs or vehicle was injected into the tail vein 7 days after STZ injection. The fasting blood glucose levels were monitored weekly. A glucose tolerance test was performed, and the parotid gland weight, salivary flow rate, oxidative stress indices, parotid gland histology, and caspase-3, vascular endothelial growth factor (VEGF), and proliferating cell nuclear antigen (PCNA) expression in parotid tissues were assessed 28 days post-transplantation. Results: Transplantation of hDPSCs downregulated blood glucose, improved the salivary flow rate, and reduced oxidative stress. The cells migrated to, survived, and differentiated into acinar, ductal, and myoepithelial cells in the STZ-injured parotid gland. Moreover, they downregulated the expression of caspase-3 and upregulated the expression of VEGF and PCNA, likely exerting pro-angiogenetic and antiapoptotic effects and promoting endogenous regeneration. In addition, the transplanted cells enhanced the parotid nitric oxide (NO) -tetrahydrobiopterin (BH4) pathway. Conclusions: Our results show that hDPSCs can migrate to and survive within the STZ-injured parotid gland, where they prevent its functional and morphological damage by restoring normal glucose levels, differentiating into parotid cell populations, and stimulating paracrine-mediated regeneration. Thus, hDPSCs may have therapeutic potential in the treatment of diabetes-induced parotid gland injury.

Keywords: dental pulp stem cells, diabetes, streptozotocin, parotid gland

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Authors: Deb Kumar Shah, M. Shaheer Akhtar, Ha Ryeon Lee, O-Bong Yang, Chong Yeal Kim

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The crystalline silicon (Si) solar cells are highly renowned photovoltaic technology and well-established as the commercial solar technology. Most of the solar panels are globally installed with the crystalline Si solar modules. At the present scenario, the major photovoltaic (PV) market is shared by c-Si solar cells, but the cost of c-Si panels are still very high as compared with the other PV technology. In order to reduce the cost of Si solar panels, few necessary steps such as low-cost Si manufacturing, cheap antireflection coating materials, inexpensive solar panel manufacturing are to be considered. It is known that the antireflection (AR) layer in c-Si solar cell is an important component to reduce Fresnel reflection for improving the overall conversion efficiency. Generally, Si wafer exhibits the 30% reflection because it normally poses the two major intrinsic drawbacks such as; the spectral mismatch loss and the high Fresnel reflection loss due to the high contrast of refractive indices between air and silicon wafer. In recent years, researchers and scientists are highly devoted to a lot of researches in the field of searching effective and low-cost AR materials. Silicon nitride (SiNx) is well-known AR materials in commercial c-Si solar cells due to its good deposition and interaction with passivated Si surfaces. However, the deposition of SiNx AR is usually performed by expensive plasma enhanced chemical vapor deposition (PECVD) process which could have several demerits like difficult handling and damaging the Si substrate by plasma when secondary electrons collide with the wafer surface for AR coating. It is very important to explore new, low cost and effective AR deposition process to cut the manufacturing cost of c-Si solar cells. One can also be realized that a nano-texturing process like the growth of nanowires, nanorods, nanopyramids, nanopillars, etc. on Si wafer can provide a low reflection on the surface of Si wafer based solar cells. The above nanostructures might be enhanced the antireflection property which provides the larger surface area and effective light trapping. In this work, we report on the development of crystalline Si solar cells without using the AR layer. The Silicon wafer was modified by growing nanowires like Si nanostructures using the wet controlled etching method and directly used for the fabrication of Si solar cell without AR. The nanostructures over Si wafer were optimized in terms of sizes, lengths, and densities by changing the etching conditions. Well-defined and aligned wires like structures were achieved when the etching time is 20 to 30 min. The prepared Si nanostructured displayed the minimum reflectance ~1.64% at 850 nm with the average reflectance of ~2.25% in the wavelength range from 400-1000 nm. The nanostructured Si wafer based solar cells achieved the comparable power conversion efficiency in comparison with c-Si solar cells with SiNx AR layer. From this study, it is confirmed that the reported method (controlled wet etching) is an easy, facile method for preparation of nanostructured like wires on Si wafer with low reflectance in the whole visible region, which has greater prospects in developing c-Si solar cells without AR layer at low cost.

Keywords: chemical etching, conversion efficiency, silicon nanostructures, silicon solar cells, surface modification

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Authors: A. Reum Son, Jin Seon Kwon, Seung Hun Park, Hai Bang Lee, Moon Suk Kim

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These days, stem cell therapy is focused on for promising source of treatment in clinical human disease. As a supporter of stem cells, in situ-forming hydrogels with growth factors and cells appear to be a promising approach in tissue engineering. To examine osteogenic differentiation of hTMSCs which is one of mesenchymal stem cells in vivo in an injectable hydrogel, we use a methoxy polyethylene glycol-polycaprolactone blockcopolymer (MPEG-PCL) solution with osteogenic factors. We synthesized MPEG-PCL hydrogel and measured viscosity to check sol-gel transition. In order to demonstrate osteogenic ability of hTMSCs, we conducted in vitro osteogenesis experiment. Then, to confirm the cell cytotoxicity, we performed WST-1 with hTMSCs and MPEG-PCL. As the result of in vitro experiment, we implanted cell and hydrogel mixture into animal model and checked degree of osteogenesis with histological analysis and amount of expression genes. Through these experimental data, MPEG-PCL hydrogel has sol-gel transition in temperature change and is biocompatible with stem cells. In histological analysis and gene expression, hTMSCs are very good source of osteogenesis with hydrogel and will use it to tissue engineering as important treatment method. hTMSCs could be a good adult stem cell source for usability of isolation and high proliferation. When hTMSCs are used as cell therapy method with in situ-formed hydrogel, they may provide various benefits like a noninvasive alternative for bone tissue engineering applications.

Keywords: injectable hydrogel, stem cell, osteogenic differentiation, tissue engineering

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Authors: Wei-Wen Hu, Yong-Zhi Zhong

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Depositing conductive polypyrrole (PPy) onto elastic polydimethylsiloxane (PDMS) substrate can obtain a highly stretchable conductive film, which can be used to construct a bioreactor to cyclically stretch and electrically stimulate surface cells. However, how to completely harvest these stimulated muscle tissue to repair damaged muscle is a challenge. To address this concern, N-isopropylacrylamide (NIPAAm), a monomer of temperature-sensitive polymer, was added during the polymerization of pyrrole on PDMS so that the resulting P(Py-co-NIPAAm)/PDMS should own both conductivity and thermo-sensitivity. Therefore, cells after stimulation can be completely harvested as cell sheets by reducing temperature. Mouse skeletal myoblast, C2C12 cells, were applied to examine our hypothesis. In electrical stimulation, C2C12 cells on P(Py-co-NIPAAm)/PDMS demonstrated the best myo-differentiation under the electric field of 1 V/cm. Regarding cyclic stretching, the strain equal to or higher than 9% can highly align C2C12 perpendicular to the stretching direction. The Western blotting experiments demonstrated that the cell sheets harvested by cooling reserved more extracellular matrix (ECM) than cells collected by the traditional trypsin digestion method. Immunostaining of myosin heavy chain protein (MHC) indicated that both mechanical and electrical stimuli effectively increased the number of myotubes and the differentiation ratio, and the myotubes can be aligned by cyclic stretching. Stimulated cell sheets can be harvested by cooling, and the alignment of myotubes was still maintained. These results suggested that the deposition of P(Py-co-NIPAAm) on PDMS can be applied to harvest intact cell sheets after cyclic stretching and electrical stimulation, which increased the feasibility of bioreactor for the application of tissue engineering and regenerative medicine.

Keywords: bioreactor, cell sheet, conductive polymer, cyclic stretching, electrical stimulation, muscle tissue engineering, myogenesis, thermosensitive hydrophobicity

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Authors: Masocorro Gawned, Stephen Myers, Guat Siew Chew

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Nuclear Receptors (NR) are a super family of transcription factors that play a major role in lipid and glucose metabolism in skeletal muscle. Recently, pharmacological evidence supports the view that stimulation of nuclear receptors alleviates Type 2 Diabetes (T2D). The orphan nuclear receptors (ONR) are members of the nuclear receptor (NR) superfamily whose ligands and physiological functions remain unknown. To date, no systematic studies have been carried out to screen for ONRs expressed in insulin resistant (IR) skeletal muscle cells. Therefore, in this study, we have established a model for IR by treating C2C12 skeletal muscle cells with insulin (10nM) for 48 hours. Western Blot analysis of phosphorylated AKT confirmed IR. Real-time quantitative polymerase chain reaction (qPCR) results highlighted key ONRs including NUR77 (NR4A1), NURR1 (NR4A2) and NOR1 (NR4A3) which have been associated with fatty acid oxidation regulation and glucose homeostasis. Increased mRNA expression levels of estrogen-related receptors (ERRs), REV-ERBα, NUR77, NURR1, NOR1, in insulin resistant C2C12 skeletal muscle cells, indicated that these ONRs could potentially play a pivotal regulatory role of insulin secretion in lipid metabolism. Taken together, this study has successfully contributed to the complete analysis of ONR in IR, and has filled in an important void in the study and treatment of T2D.

Keywords: type 2 diabetes, orphan nuclear receptors, transcription receptors, quantitative mRNA expression

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Authors: A. Kianifar, M. Afzali, I. Pishbin

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In this paper, design details, theoretical analysis and thermal performance analysis of a solar energy concentrator suited to combined heat and thermoelectric power generation are presented. The thermoelectric device is attached to the absorber plate to convert concentrated solar energy directly into electric energy at the focus of the concentrator. A cooling channel (water cooled heat sink) is fitted to the cold side of the thermoelectric device to remove the waste heat and maintain a high temperature gradient across the device to improve conversion efficiency.

Keywords: concentrator thermoelectric generator, CTEG, solar energy, thermoelectric cells

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Authors: Magdalena Hałupka-Bryl, Magdalena Bednarowicz, Ryszard Krzyminiewski, Yukio Nagasaki

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Due to their unique physical properties, superparamagnetic iron oxide nanoparticles are increasingly used in medical applications. They are very useful carriers for delivering antitumor drugs in targeted cancer treatment. Magnetic nanoparticles (PEG-PIONs/DOX) with chemotherapeutic were synthesized by coprecipitation method followed by coating with biocompatible polymer PEG-derivative (poly(ethylene glycol)-block-poly(4-vinylbenzylphosphonate). Complete physicochemical characterization was carried out (ESR, HRTEM, X-ray diffraction, SQUID analysis) to evaluate the magnetic properties of obtained PEG-PIONs/DOX. Nanoparticles were investigated also in terms of their stability, drug loading efficiency, drug release and antiproliferative effect on cancer cells. PEG-PIONs/DOX have been successfully used for the efficient delivery of an anticancer drug into the tumor region. Fluorescent imaging showed the internalization of PEG-PIONs/DOX in the cytoplasm. Biodistribution studies demonstrated that PEG-PIONs/DOX preferentially accumulate in tumor region via the enhanced permeability and retention effect. The present findings show that synthesized nanosystem is promising tool for potential magnetic drug delivery.

Keywords: targeted drug delivery, magnetic properties, iron oxide nanoparticles, biodistribution

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Authors: Chhabi Nigam, S. Ramakrishnan

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This paper brings out the analysis of the airborne Synthetic Aperture Radar (SAR) data to study the impact of Doppler centroid on Image quality and geocoding accuracy from the perspective of Stripmap mode of data acquisition. Although in Stripmap mode of data acquisition radar beam points at 90 degrees broad side (side looking), shift in the Doppler centroid is invariable due to platform motion. In-accurate estimation of Doppler centroid leads to poor image quality and image miss-registration. The effect of Doppler centroid is analyzed in this paper using multiple sets of data collected from airborne platform. Occurrences of ghost (ambiguous) targets and their power levels have been analyzed that impacts appropriate choice of PRF. Effect of aircraft attitudes (roll, pitch and yaw) on the Doppler centroid is also analyzed with the collected data sets. Various stages of the RDA (Range Doppler Algorithm) algorithm used for image formation in Stripmap mode, range compression, Doppler centroid estimation, azimuth compression, range cell migration correction are analyzed to find the performance limits and the dependence of the imaging geometry on the final image. The ability of Doppler centroid estimation to enhance the imaging accuracy for registration are also illustrated in this paper. The paper also tries to bring out the processing of low squint SAR data, the challenges and the performance limits imposed by the imaging geometry and the platform dynamics on the final image quality metrics. Finally, the effect on various terrain types, including land, water and bright scatters is also presented.

Keywords: ambiguous target, Doppler Centroid, image registration, Airborne SAR

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Authors: Ethan James

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Over one billion people worldwide suffer from some level of vision loss or blindness as a result of progressive retinal diseases. Many patients, particularly in developing areas, are incorrectly diagnosed or undiagnosed whatsoever due to unconventional diagnostic tools and screening methods. Artificial intelligence (AI) based on deep learning (DL) convolutional neural networks (CNN) have recently gained a high interest in ophthalmology for its computer-imaging diagnosis, disease prognosis, and risk assessment. Optical coherence tomography (OCT) is a popular imaging technique used to capture high-resolution cross-sections of retinas. In ophthalmology, DL has been applied to fundus photographs, optical coherence tomography, and visual fields, achieving robust classification performance in the detection of various retinal diseases including macular degeneration, diabetic retinopathy, and retinitis pigmentosa. However, there is no complete diagnostic model to analyze these retinal images that provide a diagnostic accuracy above 90%. Thus, the purpose of this project was to develop an AI model that utilizes machine learning techniques to automatically diagnose specific retinal diseases from OCT scans. The algorithm consists of neural network architecture that was trained from a dataset of over 20,000 real-world OCT images to train the robust model to utilize residual neural networks with cyclic pooling. This DL model can ultimately aid ophthalmologists in diagnosing patients with these retinal diseases more quickly and more accurately, therefore facilitating earlier treatment, which results in improved post-treatment outcomes.

Keywords: artificial intelligence, deep learning, imaging, medical devices, ophthalmic devices, ophthalmology, retina

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Authors: Fentie Bishaw Wagayie

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This study was carried out with the objective of studying the response of Farta sheep in feed intake and live weight change when fed on hay supplemented with noug seed cake (NSC), wheat bran (WB), and their mixtures. The digestibility trial of 7 days and 90 days of feeding trial was conducted using 25 intact male Farta sheep with a mean initial live weight of 16.83 ± 0.169 kg. The experimental animals were arranged randomly into five blocks based on the initial live weight, and the five treatments were assigned randomly to each animal in a block. Five dietary treatments used in the experiment comprised of grass hay fed ad libitum (T1), grass hay ad libitum + 300 g DM WB (T2), grass hay ad libitum + 300 g DM (67% WB: 33% NSC mixture) (T3), grass hay ad libitum + 300 g DM (67% NSC: 33% WB) (T4) and 300 g DM/ head/day NSC (T5). Common salt and water were offered ad libitum. The supplements were offered twice daily at 0800 and 1600 hours. The experimental sheep were kept in individual pens. Supplementation of NSC, WB, and their mixtures significantly increased (p < 0.01) the total dry matter (DM) (665.84-788 g/head/day) and (p < 0.001) crude protein (CP) intake. Unsupplemented sheep consumed significantly higher (p < 0.01) grass hay DM (540.5g/head/day) as compared to the supplemented treatments (365.8-488 g/h/d), except T2. Among supplemented sheep, T5 had significantly higher (p < 0.001) CP intake (99.98 g/head/day) than the others (85.52-90.2 g/head/day). Supplementation significantly improved (p < 0.001) the digestibility of CP (66.61-78.9%), but there was no significant effect (p > 0.05) on DM, OM, NDF, and ADF digestibility between supplemented and control treatments. Very low CP digestibility (11.55%) observed in the basal diet (grass hay) used in this study indicated that feeding sole grass hay could not provide nutrients even for the maintenance requirement of growing sheep. Significant final and daily live weight gain (p < 0.001) in the range of 70.11-82.44 g/head/day was observed in supplemented Farta sheep, but unsupplemented sheep lost weight by 9.11g/head/day. Numerically, among the supplemented treatments, sheep supplemented with a higher proportion of NSC in T4 (201 NSC + 99 g WB) gained more weight than the rest, though not statistically significant (p > 0.05). The absence of statistical difference in daily body weight gain between all supplemented sheep indicated that the supplementation of NSC, WB, and their mixtures had similar potential to provide nutrients. Generally, supplementation of NSC, WB, and their mixtures to the basal grass hay diet improved feed conversion ratio, total DM intake, CP intake, and CP digestibility, and it also improved the growth performance with a similar trend for all supplemented Farta sheep over the control group. Therefore, from a biological point of view, to attain the required level of slaughter body weight within a short period of the growing program, sheep producer can use all the supplement types depending upon their local availability, but in the order of priority, T4, T5, T3, and T2, respectively. However, based on partial budget analysis, supplementation of 300 g DM/head /day NSC (T5) could be recommended as profitable for producers with no capital limitation, whereas T4 supplementation (201 g NSC + 99 WB DM/day) is recommended when there is capital scarcity.

Keywords: weight gain, supplement, Farta sheep, hay as basal diet

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Authors: Ebru Ergi̇n, Funda Kutlu

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Emerging adulthood is a developmental period and the formation of identity is crucial task of emerging adults in this period. In this frame, the main aim of the study was to compare college students and full-time workers on emerging adulthood dimensions and identity statuses in relation to some demographic variables in Turkey. The participants of the study were university students studying in Ankara and the employees working full-time in Ankara and Bursa. The mean age of the sample was 20.84 (sd=1.84), ranging from 18 to 25. The measurement instruments of the study were Inventory of Dimensions of Emerging Adulthood and Extended Objective Measure of Ego Identity Status (EOMEIS-II). The participants’ data (N=313) were analyzed to test the research questions and hypotheses of the study. A series of MANOVA were performed to test the group differences for some demographic characteristics (such as: employee/student, male/female, living with family/living apart from family) on scores of emerging adulthood dimensions and identity status. The results of the MANOVAs indicated that students, females and participants who live apart from their families had higher scores on emerging adulthood dimensions. The results of the identity status scores differences depending on the demographic characteristic pointed out that there were a significant group differences for identity foreclosure identity scores between employees and students. Employees’ foreclosure identity scores were higher than students. Furthermore, the identity scores were differed significantly according to gender of the participants. Male participants had higher scores in moratorium and foreclosure identity and female participants have higher achievement identity scores than males. Also, the participants who live with their family scored higher in foreclosure identity and the participants who live apart from their family scored higher in identity achievement status.

Keywords: college students, emerging adulthood, full-time employees, identity statuses

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Authors: Merry Meryam Martgrita, Marselina Irasonia Tan

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One of several aggresive cancer is cancer that overexpress c-erbB2 receptor along with the expression of estrogen receptor. Components of extracellular matrix play an important role to increase cancer cells proliferation, migration and invasion. Both components can affect cancer development by regulating the signal transduction pathways in cancer cells. In recent research, SKOV-3 ovarian cancer cell line, that overexpress c-erbB2 receptor was cultured on type IV collagen and treated with estradiol-17β, to reveal the role of both components on RNA and protein level of c-erbB2 receptor. In this research we found a modulation phenomena of increasing and decreasing of c-erbB2 RNA level and a stabilisation phenomena of c-erbB2 protein expression due to estradiol-17β and type IV collagen. It seemed that estradiol-17β has an important role to increase c-erbB2 transcription and the stability of c-erbB2 protein expression. Type IV collagen has an opposite role. It blocked c-erbB2 transcription when it bound to integrin receptor in SKOV-3 cells.

Keywords: c-erbB2, estradiol-17β, SKOV-3, type IV collagen

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Authors: Ja’Afar Umar, Naziru Salisu

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The consumption of powdered or industrialized juices has increased globally due to the fast pace of city life. These foods, with their attractive color, odor, and taste, are easily diluted in water and can lead to obesity, diabetes, hypertension, and cardiovascular problems. In a study, 80 purple varieties of onion bulbs were used to evaluate the cytotoxicity of the Tiara and Bevi mix beverage powder. The viability of the bulbs was tested using the A. cepa toxicity test. The bulbs were divided into five groups, and the root growth was recorded. The mixture was then squashed in a 45% acetic acid solution and examined for chromosomal abnormalities. The chromosomal abnormalities were classified as bridges, c-mitoses, vagrants, fragments, stickiness, bi-nuclei, and multi-polar. The study found that the highest number of dividing cells was in the negative control group, followed by the group treated with BM beverage. The highest number of aberrant cells was in the group treated with TR beverage, followed by BM 5%. Stickiness of cells was observed in both BM and TR 5% beverage concentrations. No lagging chromosome was present in the negative control group. The highest mitotic index was in the negative control group, and bridge fragrance was observed in the groups treated with different beverages. This study highlights the importance of Allium cepa L. in genotoxic substance testing, revealing chromosomal and mitotic abnormalities in root tip cells. The study also reveals that at 5% concentrations, root growth decreases, indicating potential genetic abnormalities in Allium cepa's genetic material.

Keywords: cytotoxicity, Allium cepa, Beverages, Chromosome

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Authors: H. Bouaziz, M. Sefi, J. de Lapuente, M. Borras, N. Zeghal

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Although arsenic trioxide has been the subject of toxicological research, in vitro cytotoxicity and genotoxicity studies using relevant cell models and uniform methodology are not well elucidated. Hence, the aim of the present study was to evaluate the cytotoxicity and genotoxicity induced by arsenic trioxide in human keratinocytes (HaCaT) using the MTT [3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] and alkaline single cell gel electrophoresis (Comet) assays, respectively. Human keratinocytes were treated with different doses of arsenic trioxide for 4 h prior to cytogenetic assessment. Data obtained from the MTT assay indicated that arsenic trioxide significantly reduced the viability of HaCaT cells in a dose-dependent manner, showing a IC50 value of 34.18 ± 0.6 µM. Data generated from the comet assay also indicated a significant dose-dependent increase in DNA damage in HaCaT cells associated with arsenic trioxide exposure. We observed a significant increase in comet tail length and tail moment, showing an evidence of arsenic trioxide -induced genotoxic damage in HaCaT cells. This study confirms that the comet assay is a sensitive and effective method to detect DNA damage caused by arsenic.

Keywords: arsenic trioxide, cytotoxixity, genotoxicity, HaCaT

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Authors: A. Aljohani, D. Burke, D. Clarke, M. Gormally, M. Byrne, G. Fleming

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Introduction: House Dust Mites (HDMs) are a source of allergen particles embedded in textiles and furnishings. Vacuum sampling is commonly used to recover and determine the abundance of HDMs but the efficiency of this method is less than standardized. Here, the efficiency of recovery of HDMs was evaluated from home-associated textiles using vacuum sampling protocols.Methods/Approach: Living Mites (LMs) or dead Mites (DMs) House Dust Mites (Dermatophagoides pteronyssinus: FERA, UK) were separately seeded onto the surfaces of Smooth Cotton, Denim and Fleece (25 mites/10x10cm2 squares) and left for 10 minutes before vacuuming. Fabrics were vacuumed (SKC Flite 2 pump) at a flow rate of 14 L/min for 60, 90 or 120 seconds and the number of mites retained by the filter (0.4μm x 37mm) unit was determined. Vacuuming was carried out in a linear direction (Protocol 1) or in a multidirectional pattern (Protocol 2). Additional fabrics with LMs were also frozen and then thawed, thereby euthanizing live mites (now termed EMs). Results/Findings: While there was significantly greater (p=0.000) recovery of mites (76% greater) in fabrics seeded with DMs than LMs irrespective of vacuuming protocol or fabric type, the efficiency of recovery of DMs (72%-76%) did not vary significantly between fabrics. For fabrics containing EMs, recovery was greatest for Smooth Cotton and Denim (65-73% recovered) and least for Fleece (15% recovered). There was no significant difference (p=0.99) between the recovery of mites across all three mite categories from Smooth Cotton and Denim but significantly fewer (p=0.000) mites were recovered from Fleece. Scanning Electron Microscopy images of HMD-seeded fabrics showed that live mites burrowed deeply into the Fleece weave which reduced their efficiency of recovery by vacuuming. Research Implications: Results presented here have implications for the recovery of HDMs by vacuuming and the choice of fabric to ameliorate HDM-dust sensitization.

Keywords: allergy, asthma, dead, fabric, fleece, live mites, sampling

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Authors: Brayden Mi

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Reverse time migration has been widely used in the Petroleum exploration industry to reveal subsurface images and to detect rock and fluid properties since the early 1980s. The seismic technology involves the construction of a velocity model through interpretive model construction, seismic tomography, or full waveform inversion, and the application of the reverse-time propagation of acquired seismic data and the original wavelet used in the acquisition. The methodology has matured from 2D, simple media to present-day to handle full 3D imaging challenges in extremely complex geological conditions. Conventional Ultrasound computed tomography (USCT) utilize travel-time-inversion to reconstruct the velocity structure of an organ. With the velocity structure, USCT data can be migrated with the “bend-ray” method, also known as migration. Its seismic application counterpart is called Kirchhoff depth migration, in which the source of reflective energy is traced by ray-tracing and summed to produce a subsurface image. It is well known that ray-tracing-based migration has severe limitations in strongly heterogeneous media and irregular acquisition geometries. Reverse time migration (RTM), on the other hand, fully accounts for the wave phenomena, including multiple arrives and turning rays due to complex velocity structure. It has the capability to fully reconstruct the image detectable in its acquisition aperture. The RTM algorithms typically require a rather accurate velocity model and demand high computing powers, and may not be applicable to real-time imaging as normally required in day-to-day medical operations. However, with the improvement of computing technology, such a computational bottleneck may not present a challenge in the near future. The present-day (RTM) algorithms are typically implemented from a flat datum for the seismic industry. It can be modified to accommodate any acquisition geometry and aperture, as long as sufficient illumination is provided. Such flexibility of RTM can be conveniently implemented for the application in USCT imaging if the spatial coordinates of the transmitters and receivers are known and enough data is collected to provide full illumination. This paper proposes an implementation of a full 3D RTM algorithm for USCT imaging to produce an accurate 3D acoustic image based on the Phase-shift-plus-interpolation (PSPI) method for wavefield extrapolation. In this method, each acquired data set (shot) is propagated back in time, and a known ultrasound wavelet is propagated forward in time, with PSPI wavefield extrapolation and a piece-wise constant velocity model of the organ (breast). The imaging condition is then applied to produce a partial image. Although each image is subject to the limitation of its own illumination aperture, the stack of multiple partial images will produce a full image of the organ, with a much-reduced noise level if compared with individual partial images.

Keywords: illumination, reverse time migration (RTM), ultrasound computed tomography (USCT), wavefield extrapolation

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Authors: Hoda M. Eid, Abir Nachar, Farah Thong, Gary Sweeney, Pierre S. Haddad

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Caffeic acid methyl ester (CAME) and caffeic ethyl esters (CAEE) were previously reported to potently stimulate glucose uptake in cultured C2C12 skeletal muscle cells via insulin-independent mechanisms involving the activation of adenosine monophosphate-activated protein kinase (AMPK). In the present study, we investigated the effect of the two compounds on the translocation of glucose transporter GLUT4 in L6 skeletal muscle cells. The cells were treated with the optimum non-toxic concentration (50 µM) of either CAME or CAEE for 18 h. Levels of GLUT4myc at the cell surface were measured by O-phenylenediamine dihydrochloride (OPD) assay. The effects of CAME and CAEE on GLUT1 and GLUT4 protein content were also measured by western immunoblot. Our results show that CAME and CAEE significantly increased glucose uptake, GLUT4 translocation and GLUT4 protein content. Furthermore, the effect of the two CA esters on two insulin-sensitive cell lines: H4IIE rat hepatoma and 3T3-L1 adipocytes were investigated. CAME and CAEE reduced the enzymatic activity of the key hepatic gluconeogenic enzyme glucose-6-phosphatase in a concentration-dependent manner. In addition, they exerted a concentration-dependent antiadipogenic effect on 3T3-L1 cells. Mitotic clonal expansion (MCE), a prerequisite for adipocytes differentiation was also concentration-dependently inhibited. The two compounds abrogated lipid droplet accumulation, blocked MCE and maintained cells in fibroblast-like state when applied at the maximum non-toxic concentration (100 µM). In addition, the expression of the early key adipogenic transcription factors CCAAT enhancer-binding protein beta (C/EBP-β) and the master regulator of adipogenesis peroxisome-proliferator-activated receptor gamma (PPAR-γ) were inhibited. We, therefore, conclude that CAME and CAEE exert pleiotropic benefits in several insulin-sensitive cell lines through insulin-independent mechanisms involving AMPK, hence they may treat obesity, diabetes and other metabolic diseases.

Keywords: type 2 diabetes mellitus, insulin resistance, GLUT4, Akt, AMPK.

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Authors: Muhammad Ali Syed, Arshad Saleem Bhatti, Chen-zhong Li, Habib Ali Bokhari

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Surface plasmon resonance (SPR) biosensors have emerged as a promising technique for bioanalysis as well as microbial detection and identification. Real time, sensitive, cost effective, and label free detection of biomolecules from complex samples is required for early and accurate diagnosis of infectious diseases. Like many other types of optical techniques, SPR biosensors may also be successfully utilized for microbial detection for accurate, point of care, and rapid results. In the present study, we have utilized a commercially available automated SPR biosensor of BI company to study the microbial detection form water samples spiked with different concentration of Staphylococcus aureus bacterial cells. The gold thin film sensor surface was functionalized to react with proteins such as protein G, which was used for directed immobilization of monoclonal antibodies against Staphylococcus aureus. The results of our work reveal that this immunosensor can be used to detect very small number of bacterial cells with higher sensitivity and specificity. In our case 10^3 cells/ml of water have been successfully detected. Therefore, it may be concluded that this technique has a strong potential to be used in microbial detection and identification.

Keywords: surface plasmon resonance (SPR), Staphylococcus aureus, biosensors, microbial detection

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Authors: Philip Speranza

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As everyday transit options are shifting from autocentric to pedestrian and bicycle oriented modes for healthy living, downtown streets are becoming more attractive places to live. However, tools and methods to measure the natural environment at the small scale of streets do not exist. Fortunately, a combination of mobile data collection technology and parametric urban design software now allows an interface to relate urban ecological conditions. This paper describes creation of an interactive tool to measure urban phenomena of air, water, and heat/light at the scale of new three-by-three block pedestrianized areas in Barcelona called Superilles. Each Superilla limits transit to the exterior of the blocks and to create more walkable and bikeable interior streets for healthy living. The research will describe the integration of data collection, analysis, and design output via a live interface using parametric software Rhino Grasshopper and the Human User Interface (UI) plugin.

Keywords: transit, urban design, GIS, parametric design, Superilles, Barcelona, urban ecology

Procedia PDF Downloads 245

Authors: Elif Gencturk, Ekin Yurdakul, Ahmet Y. Celik, Senol Mutlu, Kutlu O. Ulgen

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Yeast cells are generally used as a model system of eukaryotes due to their complex genetic structure, rapid growth ability in optimum conditions, easy replication and well-defined genetic system properties. Thus, yeast cells increased the knowledge of the principal pathways in humans. During fermentation, carbohydrates (hexoses and pentoses) degrade into some toxic by-products such as 5-hydroxymethylfurfural (5-HMF or HMF) and furfural. HMF influences the ethanol yield, and ethanol productivity; it interferes with microbial growth and is considered as a potent inhibitor of bioethanol production. In this study, yeast single cell behavior under HMF application was monitored by using a continuous flow single phase microfluidic platform. Microfluidic device in operation is fabricated by hot embossing and thermo-compression techniques from cyclo-olefin polymer (COP). COP is biocompatible, transparent and rigid material and it is suitable for observing fluorescence of cells considering its low auto-fluorescence characteristic. The response of yeast cells was recorded through Red Fluorescent Protein (RFP) tagged Nop56 gene product, which is an essential evolutionary-conserved nucleolar protein, and also a member of the box C/D snoRNP complexes. With the application of HMF, yeast cell proliferation continued but HMF slowed down the cell growth, and after HMF treatment the cell proliferation stopped. By the addition of fresh nutrient medium, the yeast cells recovered after 6 hours of HMF exposure. Thus, HMF application suppresses normal functioning of cell cycle but it does not cause cells to die. The monitoring of Nop56 expression phases of the individual cells shed light on the protein and ribosome synthesis cycles along with their link to growth. Further computational study revealed that the mechanisms underlying the inhibitory or inductive effects of HMF on growth are enriched in functional categories of protein degradation, protein processing, DNA repair and multidrug resistance. The present microfluidic device can successfully be used for studying the effects of inhibitory agents on growth by single cell tracking, thus capturing cell to cell variations. By metabolic engineering techniques, engineered strains can be developed, and the metabolic network of the microorganism can thus be manipulated such that chemical overproduction of target metabolite is achieved along with the maximum growth/biomass yield.  

Keywords: COP, HMF, ribosome biogenesis, thermoplastic microbioreactor, yeast

Procedia PDF Downloads 169

Authors: Emanuela Chiarella, Clelia Nisticò, Nicola Lombardo, Giovanna Lucia Piazzetta, Nadia Lobello, Maria Mesuraca

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Killian’s Antrochoanal Polyp is a benign lesion of the maxillary sinus characterized by unilateral nasal obstruction, pus discharge, and headache. It affects, more commonly children and young adults. Although its etiology still remains unclear, chronic inflammation, autoreactivity, allergies, and viral infections are strongly associated with its formation and development, resulting in nasal tissue remodeling. We aimed to investigate the stem cells components which reside in this pathological tissue. In particular, we adopted a protocol for the isolation and culturing of mesenchymal stem cells from surgical biopsies of three Killian nasal polyp patients (KNP-MSCs) as well as from their healthy nasal tissue (HNT-MSCs) that were used as controls. The immunophenotype profile of HNT-MSCs and KNP-MSCs was more similar, with a marked positivity for CD73, CD90, and CD105 expression, while being negative for CD34 and CD14 haematopoietic genes. Cell proliferation assay showed that KNP-MSCs had a replicative disadvantage compared to HNT-MSCs, as evidenced by the significantly lower number of cells in the S-phase of the cell cycle. KNP-MSCs also took longer to close a wound than HNT-MSCs, indicating a partial epithelial phenotype in which low levels of ICAM-1 mRNA and a significant increase in E-CAD transcript were detectable. Subsequently, the differentiation potential of both MSCs populations was analyzed by inducing osteoblastic or adipocyte differentiation for up to 20 days. KNP-MSCs showed the ability to differentiate into osteoblasts, although ALP activity as well as the number and size of calcium deposits were lower than osteogenic induced-HNT-MSCs. Also, mRNA levels of osteoblastic marker genes (OCN, OPN, OSX, RUNX2) resulted lower compared to control cell population. Instead, the analysis of the adipogenic differentiation potential showed a similar behavior between KNP-MSCs and HNT-MSCs considering that the amount of lipid droplets, the expression of adipocyte-specific genes (FABP4, AdipoQ, PPARγ2, LPL) and the content of triacylglycerols were almost overlapping. Taken together, these results first demonstrated that Killian's nasal polyp is a source of mesenchymal stem cells with self-renewal and multi-differentiative capabilities.

Keywords: Mesenchymal stem cells, adipogenic differentiation, osteogenic differentiation, EMT

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Authors: A. Vijayendra Chary, R. Hemalatha

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Background: Vitamin D is a potent modulator of the immune system and is involved in regulating cell proliferation and differentiation. Vitamin D deficiency has been linked to increased severity of asthma in children. Asthma has dramatically increased in past decades, particular in developing countries and affects up to 20% of the population. IgE and its receptors, CD23 (FcεRII) and CD 21, play an essential role in all allergic conditions. Methods: A case control study was conducted on asthma and age and sex matched control children. 25 hydroxyvitamin D3 was quantified by HPLC; CD23; and CD21 expression on B cells were performed by flow cytometry. Total Histamine, total IGE and IL-5 and IFN-γ cytokines were determined by ELISA in blood samples of bronchial asthma (n=45) and control children (n=45). Results: The mean ± SE of vitamin D was significantly (p<0.05) low in asthma children (13.6±0.54 ng/mL) than in controls (17.4 ± 0.37 ng/mL). The mean (%) ± SE of CD23 and CD21 expression on B cells were significantly (p<0.01) high in asthma (1.02±0.09; 1.67± 0.13), when compared to controls (0.24±0.01; 0.94±0.03) respectively. The mean± SE of Serum IgE and blood histamine levels in asthma children (354.52 ± 17.33 IU/mL; 53.27 ± 2.54 nM/mL) were increased (P<0.05) when compared to controls (183.12±17.62 IU/mL 39.34±4.16 nM/mL) respectively and IFN-γ (Th1 cytokine) was lower (P<0.01) (16.37±1.27 pg/mL) than in controls (43.34±6.21 pg/mL). Conclusion: Our study provides evidence that low vitamin D levels are associated with increased IgE receptors CD23 and CD21 on B cells. In addition, there was preferential activation of Th2 (IL-5) and suppression of Th1 (IFN-γ) cytokines in children with asthma.

Keywords: bronchial asthma, CD23, IgE, vitamin D

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Authors: Gupta Renu, T. S. Roy

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Introduction: For the prospect of successful replacement therapies in treatment of Diabetes mallitus it is necessary to know events occurring during normal human pancreas development. Literature of human pancreas development are few in number as well as mainly related to first trimester because of ethical and technical difficulties. So the study was conducted on 12 fetuses from 12 gestational weeks (GW) to 5 months of infant to know normal development of exocrine and endocrine part of human pancreas. Material and Methods: Human fetalpancreases were screened by haematoxyline and eosin staining and done electron microscopy for suitable specimens to know ultrastructural detail of fetal pancreas. Results:It was observed arborized tubules, the cells budding out from these tubules differentiated into primitive acini and islets in 12thGW. At 14 weeks scanty granules were observed in the endocrine cells which coincided with the capillary invasion of the islets. The ducts and acini were surrounded by well-organized connective tissue. The acinihad elongated cells, small amount of cytoplasm and large open face euchromatic nuclei with single nucleolus. The mature form of islets of Langerhans was observed close to the acini and duct in 20 GW fetus. Connective tissue around the duct was well organized.No significant developmental change was observed early postnatal, infant. Conclusion: The development of both component exocrine as well as endocrine part of human fetal pancreas was studied by light and electron microscopy. Observations suggested that the fetal pancreas contained mainly ducts, few acini, many centroacinar cells, and large undifferentiated tissue.

Keywords: gestational weeks (GW), acini, islets of Langerhans, ducts

Procedia PDF Downloads 262

Authors: Prajna Paramita Naik, Sujit Kumar Bhutia

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Background: Regardless of the development multimodal treatment strategies, oral squamous cell carcinoma (OSCC) is often associated with a high rate of recurrence, metastasis and chemo- and radio- resistance. The present study inspected the relevance of CD44, ABCB1 and ADAM17 expression as a putative stem cell compartment in oral squamous cell carcinoma (OSCC) and deciphered the role of autophagy in regulating the expression of aforementioned proteins, stemness and chemoresistance. Methods: A retrospective analysis of CD44, ABCB1 and ADAM17 expression with respect to the various clinicopathological factors of sixty OSCC patients were determined via immunohistochemistry. The correlation among CD44, ABCB1 and ADAM17 expression was established. Sphere formation assay, flow cytometry and fluorescence microscopy were conducted to elucidate the stemness and chemoresistance nature of established cisplatin-resistant oral cancer cells (FaDu). The pattern of expression of CD44, ABCB1 and ADAM17 in parental (FaDu-P) and resistant FaDu cells (FaDu-CDDP-R) were investigated through fluorescence microscopy. Western blot analysis of autophagy marker proteins was performed to compare the status of autophagy in parental and resistant FaDu cell. To investigate the role of autophagy in chemoresistance and stemness, sphere formation assay, immunofluorescence and Western blot analysis was performed post transfection with siATG14 and the level of expression of autophagic proteins, mitochondrial protein and stemness-associated proteins were analyzed. The statistical analysis was performed by GraphPad Prism 4.0 software. p-value was defined as follows: not significant (n.s.): p > 0.05;*: p ≤ 0.05; **: p ≤ 0.01; ***: p ≤ 0.001; ****: p ≤ 0.0001 were considered statistically significant. Results: In OSCC, high CD44, ABCB1 and ADAM17 expression were significantly correlated with higher tumor grades and poor differentiation. However, the expression of these proteins was not related to the age and sex of OSCC patients. Moreover, the expression of CD44, ABCB1 and ADAM17 were positively correlated with each other. In vitro and OSCC tissue double labeling experiment data showed that CD44+ cells were highly associated with ABCB1 and ADAM17 expression. Further, FaDu-CDDP-R cells showed higher sphere forming capacity along with increased fraction of the CD44+ population and β-catenin expression FaDu-CDDP-R cells also showed accelerated expression of CD44, ABCB1 and ADAM17. A comparatively higher autophagic flux was observed in FaDu-CDDP-R against FaDu-P cells. The expression of mitochondrial proteins was noticeably reduced in resistant cells as compared to parental cells indicating the occurrence of autophagy-mediated mitochondrial degradation in oral cancer. Moreover, inhibition of autophagy was coupled with the decreased formation of orospheres suggesting autophagy-mediated stemness in oral cancer. Blockade of autophagy was also found to induce the restoration of mitochondrial proteins in FaDu-CDDP-R cells indicating the involvement of mitophagy in chemoresistance. Furthermore, a reduced expression of CD44, ABCB1 and ADAM17 was also observed in ATG14 deficient cells FaDu-P and FaDu-CDDP-R cells. Conclusion: The CD44+ ⁄ABCB1+ ⁄ADAM17+ expression in OSCC might be associated with chemoresistance and a putative CSC compartment. Further, the present study highlights the contribution of mitophagy in chemoresistance and confirms the potential involvement of autophagic regulation in acquisition of stem-like characteristics in OSCC.

Keywords: ABCB1, ADAM17, autophagy, CD44, chemoresistance, mitophagy, OSCC, stemness

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