Search results for: fluorescent staining

Authors: M. Klimova, O. Semyachkina-Glushkovskaya, J. Kurts, E. Zinchenko, N. Navolokin, A. Shirokov, A. Dubrovsky, A. Abdurashitov, A. Terskov, A. Mamedova, I. Agranovich, T. Antonova, I. Blokhina

Abstract:

The direction for research of Alzheimer's disease is to find an effective non-invasive and non-pharmacological way of treatment. Here we tested our hypothesis that the opening of the blood-brain barrier (BBB) induces activation of lymphatic drainage and clearing functions that can be used as a method for non-invasive stimulation of clearance of beta-amyloid and therapy of Alzheimer’s disease (AD). To test our hypothesis, in this study on healthy male mice we analyzed the interaction between BBB opening by repeated loud music (100-10000 Hz, 100 dB, duration 2 h: 60 sec – sound; 60 sec - pause) and functional changes in the meningeal lymphatic vessels (MLVs). We demonstrate clearance of dextran 70 kDa (i.v. injection), fluorescent beta-amyloid (intrahippocampal injection) and gold nanorods (intracortical injection) via MLV that significantly increased after the opening of BBB. Our studies also demonstrate that the BBB opening was associated with the improvement of neurocognitive status in mice with AD. Thus, we uncover therapeutic effects of BBB opening by loud music, such as non-invasive stimulation of lymphatic clearance of beta-amyloid in mice with AD, accompanied by improvement of their neurocognitive status. Our data are consistent with other results suggesting the therapeutic effect of BBB opening by focused ultrasound without drugs for patients with AD. This research was supported by a grant from RSF 18-75-10033

Keywords: Alzheimer's disease, beta-amyloid, blood-brain barrier, meningeal lymphatic vessels, repeated loud music

Procedia PDF Downloads 113

Authors: Samuel Mejorado

Abstract:

C. Reinahrdtii serves as one of the most promising organisms from which to obtain biological hydrogen. However, its production catalyst, [FeFe]-hydrogenase, is largely inhibited by the presence of oxygen. In recent years, researchers have identified a Proton Gradient Regulation 5 (PGR5) deficient mutant, which shows enhanced respiration and lower accumulations of oxygen within the system. In this research, we investigated the effects of varying nutrient conditions on PGR5 mutants' ability to produce hydrogen. After growing PGR5 mutants in varying nutrient conditions under 55W fluorescent lamps at 30℃ with constant stirring at 200 rpm, a common water displacement method was utilized to obtain a definitive volumetric reading of hydrogen produced by these mutants over a period of 12 days. After the trials, statistical t-tests and ANOVAs were performed to better determine the effect which nutrient conditions have on PGR5 mutants' ability to produce hydrogen. In this, we report that conditions of sulfur deprivation most optimally enhanced hydrogen production within these mutants, with groups grown under these conditions demonstrating the highest production capacity over the entire 12-day period. Similarly, it was found that when grown under conditions of nitrogen deprivation, a favorable shift towards carbon fixation and overall lipid/starch metabolism was observed. Overall, these results demonstrate that PGR5-deficient mutants stand as a promising source of biohydrogen when grown under conditions of sulfur deprivation. To date, photochemical characteristics of [FeFe]-hydrogenase in these mutants have yet to be investigated under conditions of sulfur deprivation.

Keywords: biofuel, biohydrogen, [FeFe]-hydrogenase, algal biofuel

Procedia PDF Downloads 114

Authors: Kang-Hyun Leem, Myung-Gyou Kim, Hye Kyung Kim

Abstract:

The prickly pear cactus (Opuntia ficus-indica) has a global distribution and have been used for medicinal benefits such as artherosclerosis, diabetes, gastritis, and hyperglycemia. However, very little information is currently available for their mechanism. The prikly pear variety Opuntia ficus-indica var. Saboten (OFS) is widely cultivated in Cheju Island, southwestern region of Korea, and used as a functional food. Present study investigated the effects of OFS on pancreatic β-cell function using pancreatic islet β cells (HIT cell). Alpha-glucosidase inhibition, glucose uptake, insulin secretion, insulin sensitivity, and pancreatic β cell proliferation were determined. The inhibitory effect of ethanol extract of OFS stem on α-glucosidase enzyme was measured in a cell free system. Glucose uptake was determined using fluorescent glucose analogue, 2-NBDG. Insulin secretion was measured by ELISA assay. Cell proliferation was measured by MTT assay. Ethanol extracts of OFS dose-dependently inhibited α-glucosidase activity as well as glucose uptake. Insulinotrophic effect of OFS extract was observed at high glucose media in pancreatic β-islet cells. Furthermore, pancreatic β cell regeneration was also observed.These results suggest that OFS mediates the antidiabetic activity mainly via α-glucosidase inhibition, glucose uptake, and improved insulin sensitivity.

Keywords: prickly pear cactus, Opuntia ficus-indica var. Saboten, pancreatic islet HIT cells, α-glucosidase, glucose uptake, insulinotrophic

Procedia PDF Downloads 442

Authors: Emad Heydarnia, Aref Sepasi, Nika Asefi, Sara Khakshournia, Javad Mohammadnejad

Abstract:

Background: Despite breakthrough therapeutics in breast cancer, it is one of the main causes of mortality among women worldwide. Thus, drug therapies for treating breast cancer have recently been developed by scientists. Metformin and Sorafenib are well-known therapeutic in breast cancer. In the present study, we combined Sorafenib and PCL-sorafenib with metformin to improve drug absorption and promote therapeutic efficiency. Methods: The MCF-7 cells were treated with Metformin, Sorafenib, or PCL-sorafenib. The growth inhibitory effect of these drugs and cell viability were assessed using MTT and flow cytometry assays, respectively. The expression of targeted genes involved in cell proliferation, signaling, and the cell cycle was measured by Real-time PCR. Results: The results showed that MCF-7 cells treated with Metformin/Sorafenib and PCL-sorafenib/Metformin co-treatment contributed to 50% viability compared to untreated group. Moreover, PI and Annexin V staining tests showed that the cells viability for Metformin/Sorafenib and PCL-sorafenib/Metformin was 38% and 17%, respectively. Furthermore, Sorafenib/Metformin and PCL-sorafenib/Metformin leads to p53 gene expression increase by which they can increase ROS, thereby decreasing GPX4 gene expression. In addition, they affected the expression of BCL2, and BAX genes and altered the cell cycle. Conclusion: Together, the combination of PCL-sorafenib/Metformin and Sorafenib/Metformin increased Sorafenib absorption at lower doses and also leads to apoptosis and oxidative stress increases in MCF-7 cells.

Keywords: breast cancer, metformin, nanotechnology, sorafenib

Procedia PDF Downloads 32

Authors: Abdul Salam Turkmani

Abstract:

The Lower Cretaceous rocks are exposed only in the mountains regions of Syria, such as the Anti- Lebanon mountain on the western side of Damascus. The lower cretaceous sequences are made up of different rocks. The upper and middle parts of the section are composed mainly of carbonate sediments and, less frequently, gypsum and anhydrite. The lower beds are mainly composed of sandstone, conglomerate and clay. Clay samples were collected from the study area, which is located about 45 km west of the city of Damascus, near the border village of Kfer Yabous and to the left of the Damascus -Beirut International Road, within the lower Cretaceous upper Aptian deposits. The properties of clay were carried out by X-ray diffraction (XRD) and, X-ray fluorescence (XRF) and Thermal Analysis (DTA-TG-DSC) techniques. The studied samples of clay were mainly composed of kaolinite, quartz, illite. Chemical analysis shows the content of SiO₂ varied between 46.06 to 73 % Al₂O₃ 14.55-26.56%, about the staining oxides (Fe₂O₃ + TiO₂), the total content is about 4.3 to 12.5%. The physical properties were determined by studying the behavior of the body before and after firing, showed low bending strength values (22.5 kg/cm²) after drying, and (about 247 kg/cm²) after firing at 1180°C, water absorption value was about 10%. The cubic thermal expansion coefficient at 1140°C is 213.77 x 10-7 /°C. All of the presented results confirm the suitability of this clay for the ceramic industry.

Keywords: anti-Lebanon, Damascus, ceramic, clay, thermal analysis, thermal expansion coefficient

Procedia PDF Downloads 158

Authors: Reza Hadjiaghaie Vafaie, Sevda Givtaj

Abstract:

Automation and control of biological samples and solutions at the microscale is a major advantage for biochemistry analysis and biological diagnostics. Despite the known potential of miniaturization in biochemistry and biomedical applications, comparatively little is known about fluid automation and control at the microscale. Here, we study the electric field effect inside a fluidic channel and proper electrode structures with different patterns proposed to form forward, reversal, and rotational flows inside the channel. The simulation results confirmed that the ac electro-thermal flow is efficient for the control and automation of high-conductive solutions. In this research, the fluid pumping and mixing effects were numerically studied by solving physic-coupled electric, temperature, hydrodynamic, and concentration fields inside a microchannel. From an experimental point of view, the electrode structures are deposited on a silicon substrate and bonded to a PDMS microchannel to form a microfluidic chip. The motions of fluorescent particles in pumping and mixing modes were captured by using a CCD camera. By measuring the frequency response of the fluid and exciting the electrodes with the proper voltage, the fluid motions (including pumping and mixing effects) are observed inside the channel through the CCD camera. Based on the results, there is good agreement between the experimental and simulation studies.

Keywords: microfluidic, nano/micro actuator, AC electrothermal, Reynolds number, micropump, micromixer, microfabrication, mass transfer, biomedical applications

Procedia PDF Downloads 48

Authors: Fatemeh Rezaei, Afsaneh Amouei, Iman Bakouei, Mahdi Sharif, Shahabeddin Sarvi, Mohammad Taghi Rahimi, Ahmad Daryani

Abstract:

Background: Rodents act as reservoir host and important potential source for many zoonotic pathogens which pose a public health risk to humans. Therefore, it is necessary to investigate the prevalence of gastrointestinal and ectoparasites among rodents. Materials and Methods: 118 Rattus rattus were captured using snap live traps. Each rat was combed with a fine tooth comb to dislodge ectoparasite and studied. Various samples were collected from feces, examined wet smear, formalin-ether method and stained with modified acid-fast staining and trichrome. Result: The overall prevalence of gastrointestinal parasites of examined rats was 75.4%. Cryptosporidium 30.5%, was the most prevalent protozoan which was followed by Giardia 20.3% and Entamoeba muris 13.5%, Trichomonas muris 10.1% and Spironucleus muris 3.3%. The prevalence of helminth egg was as following Syphacia obvelata 24.5%, Hymenolepis diminuta 10.1% and Trichuris muris 9.3%. 86.4% rodents were found to be infested with ectoparasites including mite 35.6%, flea 28.4%, and lice 42.7%. A significant statistical difference was observed between prevalence and gender of infected individuals. Conclusions: The prevalence of gastrointestinal and ectoparasites of collected rats in studied area is remarkably high. In addition, Rattus rattus can be considered as potential risk for human health.

Keywords: prevalence, rodent, intestinal parasites, ecto-parasites, zoonose

Procedia PDF Downloads 496

Authors: Abujnah Dukali

Abstract:

Natural honey was assessed in cell culture system for its anticancer activity. Human leukemic cell line HL 60 was treated with honey and cultured for 5 days and cytotoxicity was calculated by MTT assay. Honey showed cytotoxicity with CC50 value of 174.20 µg/ml. Radical modulation activities was assessed by lipid peroxidation assay using egg lecithin. Honey showed antioxidant activity with EC50 value of 159.73 µg/ml. In addition, treatment with HL60 cells also resulted in nuclear DNA fragmentation, as seen in agarose gel electrophoresis. This is a hallmark of cells undergoing apoptosis. Confirmation of apoptosis was performed by staining the cells with Annexin V and FACS analysis. Apoptosis is an active, genetically regulated disassembly of the cell form within. Disassembly creates changes in the phospholipid content of the cytoplasmic membrane outer leaflet. Phosphatidylserine (PS) is translocated from the inner to the outer surface of the cell for phagocytic cell recognition. The human anticoagulant, annexin V, is a Ca2+-dependent phospholipid protein with a high affinity for PS. Annexin V labeled with fluorescein can identify apoptotic cells in the population It is a confirmatory test for apoptosis. Annexin V-positive cells were defined as apoptotic cells. Since honey shows both antioxidant activity and cytotoxicity at almost the same concentration, it can prevent the free radical induced cancer as prophylactic agent and kill the cancer cells by apoptotic process as a chemotherapeutic agent. Everyday intake of honey can prevent the cancer induction.

Keywords: anticancer, cells, DNA, honey

Procedia PDF Downloads 180

Authors: Isaac Quiros-Fernandez, Angel Cid-Arregui

Abstract:

Tumor-reactive T cell receptors (TCRs) are being subject of intense investigation since they offer great potential in adoptive cell therapies against cancer. However, the identification of tumor-specific TCRs has proven challenging, for instance, due to the limited expansion capacity of tumor-infiltrating T cells (TILs) and the extremely low frequencies of tumor-reactive T cells in the repertoire of patients and healthy donors. We have developed an approach for rapid identification and characterization of neoepitope-reactive TCRs from the T cell repertoire of healthy donors. CD8 T cells isolated from multiple donors are subjected to a first sorting step after staining with HLA multimers carrying the peptide of interest. The isolated cells are expanded for two weeks, after which a second sorting is performed using the same peptide-HLA multimers. The cells isolated in this way are then processed for single-cell sequencing of their TCR alpha and beta chains. Newly identified TCRs are cloned in appropriate expression vectors for functional analysis on Jurkat, NK92, and primary CD8 T cells and tumor cells expressing the appropriate antigen. We have identified TCRs specifically binding HLA-A2 presenting epitopes of tumor antigens, which are capable of inducing TCR-mediated cell activation and cytotoxicity in target cancer cell lines. This method allows the identification of tumor-reactive TCRs in about two to three weeks, starting from peripheral blood samples of readily available healthy donors.

Keywords: cancer, TCR, tumor antigens, immunotherapy

Procedia PDF Downloads 38

Authors: Samkeliso S. Ndzimandze, Welldone Moyo, Oranso T. Mahlangu, Adolph A. Muleja, Alex T. Kuvarega, Thabo T. I. Nkambule

Abstract:

The high cost of precursor materials has hindered the commercialization of ceramic membrane technology in water treatment. In this work, a ceramic membrane disc (approximately 50 mm in diameter and 4 mm thick) was prepared from low-cost starting materials, kaolin, and fly ash by pressing at 200 bar and calcining at 900 °C. The fabricated membrane was characterized for various physicochemical properties, natural organic matter (NOM) removal as well as fouling propensity using several techniques. Further, the ceramic membrane was tested on samples collected from four drinking water treatment plants in KwaZulu-Natal, South Africa (named plants 1-4). The membrane achieved 48.6%, 54.6%, 57.4%, and 76.4% bulk UV254 reduction for raw water at plants 1, 2, 3, and 4, respectively. These removal rates were comparable to UV254 reduction achieved by coagulation/flocculation steps at the respective plants. Further, the membrane outperformed sand filtration steps in plants 1-4 in removing disinfection by-product precursors (8%-32%) through size exclusion. Fluorescence excitation-emission matrices (FEEM) studies showed the removal of fluorescent NOM fractions present in the water samples by the membrane. The membrane was fabricated using an up-scalable facile method, and it has the potential for application as a polishing step to complement conventional processes in water treatment for drinking purposes.

Keywords: crossflow filtration, drinking water treatment plants, fluorescence excitation-emission matrices, ultraviolet 254 (UV₂₅₄)

Procedia PDF Downloads 15

Authors: L. Spitaleri, C. M. A. Gangemi, R. Purrello, G. Nicotra, G. Trusso Sfrazzetto, G. Casella, M. Casarin, A. Gulino

Abstract:

Hybrid molecular–nanoparticle materials, obtained with a bottom-up approach, are suitable for the fabrication of functional nanostructures showing structural control and well-defined properties, i.e., optical, electronic or catalytic properties, in the perspective of applications in different fields of nanotechnology. Gold nanoparticles (Au NPs) exhibit important chemical, electronic and optical properties due to their size, shape and electronic structures. In fact, Au NPs containing no more than 30-40 atoms are only luminescent because they can be considered as large molecules with discrete energy levels, while nano-sized Au NPs only show the surface plasmon resonance. Hence, it appears that gold nanoparticles can alternatively be luminescent or plasmonic, and this represents a severe constraint for their use as an optical material. The aim of this work was the fabrication of nanoscale assembly of Au NPs covalently anchored to each other by means of novel bi-functional porphyrin molecules that work as bridges between different gold nanoparticles. This functional architecture shows a strong surface plasmon due to the Au nanoparticles and a strong luminescence signal coming from porphyrin molecules, thus, behaving like an artificial organized plasmonic and fluorescent network. The self-assembly geometry of this porphyrin on the Au NPs was studied by investigation of the conformational properties of the porphyrin derivative at the DFT level. The morphology, electronic structure and optical properties of the conjugated Au NPs – porphyrin system were investigated by TEM, XPS, UV–vis and Luminescence. The present nanostructures can be used for plasmon-enhanced fluorescence, photocatalysis, nonlinear optics, etc., under atmospheric conditions since our system is not reactive to air nor water and does not need to be stored in a vacuum or inert gas.

Keywords: gold nanoparticle, porphyrin, surface plasmon resonance, luminescence, nanostructures

Procedia PDF Downloads 130

Authors: Vitheejongjaroen P., Jaisin Y., Pachekrepapol U., Taweechotipatr M.

Abstract:

Lactic acid bacteria (LAB) are the most common type of microorganisms that had been used as probiotics also known for many beneficial health effects. The antioxidant activity of LAB is associated with numerous health-protective effects. This research aimed to investigate the antioxidant activity of lactic acid bacteria isolated from Thai sour pork sausage for their application in fermented milk products. Antioxidant activity determined by DPPH (2,2-diphenyl-1-picrylhydrazyl) radical scavenging assay showed that the isolate FN33-7, as 1 of 8 isolated exhibited scavenging activity in intact cell 5-7%, and supernatant 13-16%, intracellular cell free extract 42-48% respectively. This isolate was identified using 16S ribosomal DNA sequence analysis as Lactobacillus plantarum. The effect of milk fermented with L. plantarum FN33-7 on microbial count, pH and syneresis was assessed during refrigerated storage period of 28 days. The strain showed increased viability, pH level decreased, while syneresis increased. These results are similar to dairy products fermented with commercial starter cultures. Additionally, microstructure analysis of fermented milk by fluorescent microscopy showed that curd structure appeared to be dense and less porous in this fermented milk than commercial yogurt. The results of this study indicated that L. plantarum FN33-7 was a good probiotic candidate to be used in cultured milk products to reduce the risk of diseases caused by oxidative stress.

Keywords: Lactobacillus plantarum, probiotics, free radical, antioxidant, oxidative stress, fermented milk products

Procedia PDF Downloads 109

Authors: M. Sunitha, B. Nithyavani, Mathew Yohannan, S. Thiruvieni Balajji, M. A. Rathi, C. Arul Raj, P. Ragavendran, V. K. Gopalkrishnan

Abstract:

Establishment of an early and reliable biomarker for ovarian carcinogenesis whose expression can be monitored through noninvasive techniques will enable early diagnosis of cancer. Carbonic anhydrases (CA) isozymes IX and XII have been suggested to play a role in oncogenic processes. In von Hippel-Lindau (VHL)-defective tumors, the cell surface transmembrane carbonic anhydrase (CA) CA XI and CA XII genes are overexpressed because of the absence of pVHL. These enzymes are involved in causing a hypoxia condition, thereby providing an environment for metastasis. Aberrant expression of the facilitative glucose transporter GLUT I is found in a wide spectrum of epithelial malignancies. Studying the mRNA expression of CA IX, CA XII and Glut I isozymes in ovarian cancer cell lines (OAW-42 and PA-1) revealed the expression of these hypoxia genes. Immunohistochemical staining of carbonic anhydrases was also performed in 40 ovarian cancer tissues. CA IX and CA XII were expressed at 540 bp and 520 bp in OAW42, PA1 in ovarian cancer cell lines. GLUT-1 was expressed at 325bp in OAW 42, PA1 genes in ovarian cancer cell lines. Immunohistochemistry revealed high to moderate levels of expression of these enzymes. The immuostaining was seen predominantly on the cell surface membrane. The study concluded that these genes CA IX, CA XII and Glut I are expressed under hypoxic condition in tumor cells. From the present results expression of CA IX, XII and Glut I may represent potential targets in ovarian cancer therapy.

Keywords: ovarian cancer, carbonic anhydrase IX, XII, Glut I, tumor markers

Procedia PDF Downloads 342

Authors: Sibel Dikmen Kucuk, Yusuf Guner

Abstract:

In recent years, petroleum-based polymers began to be limited due to the effects on the human and environmental point of view in many countries. Thus, organic-based biodegradable materials have attracted much interest in the composite industry because of environmental concerns. As a result of this, it has been asked that inorganic and petroleum-based materials should be reduced and altered with biodegradable materials. In this point, in this study, it is aimed to investigate the potential of the use of TEMPO (2,2,6,6- tetramethylpiperidine 1-oxyl)-mediated oxidation nano-fibrillated cellulose instead of EPDM (ethylene-propylene-diene monomer) rubber, which is a petroleum-based material. Thus, the exchange of petroleum-based EPDM rubber with organic-based cellulose nanofibers, which are environmentally friendly (green) and biodegradable, will be realized. The effect of tempo-oxidized cellulose nanofibers (TCNF) instead of EPDM rubber was analyzed by rheological, mechanical, chemical, thermal, and aging analyses. The aged surfaces were visually scrutinized, and surface morphological changes were examined via scanning electron microscopy (SEM). The results obtained showed that TEMPO oxidation nano-fibrillated cellulose could be used at an amount of 1.0 and 2.2 phr resulting the values stay within tolerance according to customer standard and without any chemical degradation, crack, color change or staining.

Keywords: EPDM, lignin, green materials, biodegradable fillers

Procedia PDF Downloads 99

Authors: Siddharth Deshpande, Nihar Masurkar, Vallerinteavide Mavelli Girish, Malan Desai, Chester Drum

Abstract:

Folding and stabilization of recombinant proteins remain a consistent challenge for industrial and therapeutic applications. Proteins derived from thermophilic bacteria often have superior expression and stability qualities. To develop a generalizable approach to protein folding and stabilization, we tested the hypothesis that wrapping a thermostable exoshell around a protein substrate would aid folding and impart thermostable qualities to the internalized substrate. To test the effect of internalizing a protein within a thermostable exoshell (tES), we tested in vitro folding and stability using green fluorescent protein (GFPuv), horseradish peroxidase (HRP) and renilla luciferase (rLuc). The 8nm interior volume of a thermostable ferritin assembly was engineered to accommodate foreign proteins and either present a positive, neutral or negative interior charge environment. We further engineered the tES complex to reversibly assemble and disassemble with pH titration. Template proteins were expressed as inclusion bodies and an in vitro folding protocol was developed that forced proteins to fold inside a single tES. Functional yield was improved 100-fold, 100-fold and 150-fold with use of tES for GFPuv, HRP and rLuc respectively and was highly dependent on the internal charge environment of the tES. After folding, functional proteins could be released from the tES folding cavity using size exclusion chromatography at pH 5.8. Internalized proteins were tested for improved stability against thermal, organic, urea and guanidine denaturation. Our results demonstrated that thermostable exoshells can efficiently refold and stabilize inactive aggregates into functional proteins.

Keywords: thermostable shell, in vitro folding, stability, functional yield

Procedia PDF Downloads 219

Authors: Cheng-Cao Sun, Shu-Jun Li, Cuili Yang, Yongyong Xi, Liang Wang, Feng Zhang, De-Jia Li

Abstract:

MicroRNAs (miRNAs) act as key regulators of multiple cancers. Hsa-miR-329 (miR-329) functions as a tumor suppressor in some malignancies. However, its role on lung cancer remains poorly understood. In this study, we investigated the role of miR-329 on the development of lung cancer. The results indicated that miR-329 was decreased in primary lung cancer tissues compared with matched adjacent normal lung tissues and very low levels were found in a non-small cell lung cancer (NSCLC) cell lines. Ectopic expression of miR-329 in lung cancer cell lines substantially repressed cell growth as evidenced by cell viability assay, colony formation assay and BrdU staining, through inhibiting cyclin D1, cyclin D2, and up-regulatiing p57(Kip2) and p21(WAF1/CIP1). In addition, miR-329 promoted NSCLC cell apoptosis, as indicated by up-regulation of key apoptosis gene cleaved caspase-3, and down-regulation of anti-apoptosis gene Bcl2. Moreover, miR-329 inhibited cellular migration and invasiveness through inhibiting matrix metalloproteinases (MMP)-7 and MMP-9. Further, oncogene MET was revealed to be a putative target of miR-329, which was inversely correlated with miR-329 expression. Furthermore, down-regulation of MET by siRNA performed similar effects to over-expression of miR-329. Collectively, our results demonstrated that miR-329 played a pivotal role in lung cancer through inhibiting cell proliferation, migration, invasion, and promoting apoptosis by targeting oncogenic MET.

Keywords: hsa-miRNA-329(miR-329), MET, non-small cell lung cancer (NSCLC), proliferation, apoptosis

Procedia PDF Downloads 379

Authors: Tej Varma Y, Debi D. Pant

Abstract:

Photophysics and rotational dynamics of the fluorescent probe, 6-methoxyquinoline (6MQ) with cationic surfactant, alkyltrimethylammonium bromide (nTAB) micelle solutions have been investigated (n = 12, 14 and 16). Absorption and emission peaks of the dye have been observed to shift at concentrations around critical micellar concentration (cmc) of nTAB compared to that of bulk solutions suggesting probe is in a lower polar environment. The probe senses changes in polarity (ET (30)) brought about by variation of surfactant chain length concentration and is invariably solubilized in the aqueous interface or palisade layer. The order of change in polarity observed was DTAB > CTAB > TTAB. The binding constant study shows that the probe binds strongest with TTAB (is of the order TTAB > CTAB > DTAB) due to deeper penetration into the micelle. The anisotropy decay for the probe in all the nTAB micelles studied have been rationalized based on a two-step model consisting of fast-restricted rotation of the probe and slow lateral diffusion of the probe in the micelle that is coupled to the overall rotation of the micelle. Fluorescence lifetime measurements of probe in the cationic micelles demonstrate the close proximity of the 6MQ to the Br - counterions. The fluorescence lifetimes of TTAB and DTAB are much shorter than in CTAB. These results indicate that 6MQ resides to a substantial degree in the head group region of the micelles. All the changes observed in the steady state fluorescence, microenvironment, fluorescence lifetimes, fluorescence anisotropy, and other calculations are in agreement with each other suggesting binding of the cationic surfactant with the neutral dye molecule.

Keywords: photophysics, chain length, ntaB, micelles

Procedia PDF Downloads 609

Authors: Hye Kyung Kim, Myung-Gyou Kim, Kang-Hyun Leem

Abstract:

It has been reported that deer antler extract has bone-strengthening activity and effectively used in bone diseases therapy. However, little is known about the cellular and molecular mechanism of this effect. The upper section, mid section, and base of the antler has been known to exhibit different biological properties. Present study investigated the effects of these three parts of deer antler extracts on bone formation and resorption. The effects of deer antler extracts (DH) on bone formation were determined by cell proliferation, alkaline phosphatase (ALP) activity, collagen synthesis, and mineralization in human osteoblastic MG-63 cells. The effect on bone resorption was determined by osteoclastogenesis from bone marrow-derived precursor cells driven by RANKL. Ethanol extracts of DH (50 ~ 100 µg/ml) dose-dependently increased cell proliferation, and upper part increased the cell proliferation by 118.4% while mid and base parts increased proliferation by 107.8% and 102.3%, respectively. ALP activity was significantly increased by upper part of the DH treatment. After enhancement of ALP activity, significant augmentation of collagen synthesis and calcification assessed by Sirus red and Alzarin red staining, respectively, was observed in upper part of the DH treatment. The effect of DH on bone resorption was not observed in all three parts of the DH. These results could provide a mechanistic explanation for the bone-strengthening effects of DH.

Keywords: alkaline phosphatase, collagen synthesis, deer antler, osteoblastic MG-63 cells

Procedia PDF Downloads 289

Authors: S. Mantzoukis, M. Gerasimou, P. Christodoulou, N. Varsamis, G. Kolliopoulou, N. Zotos

Abstract:

Purpose: Patients in Intensive Care Units (ICU) are exposed to a different spectrum of microorganisms relative to the hospital. Due to the fact that the majority of these patients are intubated, bronchial secretions should be examined. Material and Method: Bronchial secretions should be taken with care so as not to be mixed with sputum or saliva. The bronchial secretions are placed in a sterile container and then inoculated into blood, Mac Conkey No2, Chocolate, Mueller Hinton, Chapman and Saboureaud agar. After this period, if any number of microbial colonies are detected, gram staining is performed and then the isolated organisms are identified by biochemical techniques in the automated Microscan system (Siemens) followed by a sensitivity test in the same system using the minimum inhibitory concentration MIC technique. The sensitivity test is verified by a Kirby Bauer test. Results: In 2017 the Laboratory of Microbiology received 365 samples of bronchial secretions from the Intensive Care Unit. 237 were found positive. S. epidermidis was identified in 1 specimen, A. baumannii in 60, K. pneumoniae in 42, P. aeruginosa in 50, C. albicans in 40, P. mirabilis in 4, E. coli in 4, S. maltophilia in 6, S. marcescens in 6, S. aureus in 12, S. pneumoniae in 1, S. haemolyticus in 4, P. fluorescens in 1, E. aerogenes in 1, E. cloacae in 5. Conclusions: The majority of ICU patients appear to be a fertile ground for the development of infections. The nature of the findings suggests that a significant part of the bacteria found comes from the unit (nosocomial infection).

Keywords: bronchial secretions, cultures, infections, intensive care units

Procedia PDF Downloads 170

Authors: S. Dikmen Kucuk, A. Tozluoglu, Y. Guner

Abstract:

In recent years, petroleum-based polymers began to be limited due to effects on human and environmental point of view in many countries. Thus, organic-based biodegradable materials have attracted much interest in the composite industry because of environmental concerns. As a result of this, it has been asked that inorganic and petroleum-based materials should be reduced and altered with biodegradable materials. In this point, in this study, it is aimed to investigate the potential of use of TEMPO (2,2,6,6- tetramethylpiperidine 1-oxyl)-mediated oxidation nano-fibrillated cellulose instead of EPDM (ethylene-propylene-diene monomer) rubber, which is a petroleum-based material. Thus, the exchange of petroleum-based EPDM rubber with organic based cellulose nanofibers, which are environmentally friendly (green) and biodegradable, will be realized. The effect of tempo-oxidized cellulose nanofibers (TCNF) instead of EPDM rubber was analyzed by rheological, mechanical, chemical, thermal and aging analyses. The aged surfaces were visually scrutinized and surface morphological changes were examined via scanning electron microscopy (SEM). The results obtained showed that TEMPO oxidation nano-fibrillated cellulose can be used at an amount of 1.0 and 2.2 phr resulting the values stay within tolerance according to customer standard and without any chemical degradation, crack, colour change or staining.

Keywords: EPDM, cellulose, green materials, nanofibrillated cellulose, TCNF, tempo-oxidized nanofiber

Procedia PDF Downloads 85

Authors: Salina Yahya Saddik

Abstract:

The present study is designed to demonstrate the Ovarian Surface Epithelial cells (OSE) Estrogen Receptor α (ERα) and Progesterone Receptor (PR) during pregnancy and estrous cycle in rat. Moreover, determination of the levels of plasma progesterone, estradiol, FSH and LH were also made. The levels of plasma progesterone, estradiol, FSH and LH concentrations were determined on days 7 (n=5), 14 (n=5), and 21(n=5) of pregnancy in three groups of rats and during the estrous cycle (n=5) using ELISA kit. Immunohistochemical method for PR and ERα expression was also made on the ovary. During pregnancy, FSH and LH remained low except at term when LH levels began to increase from 16 ng/ml to 47 ng/ml. Progesterone levels significantly exceeded estradiol values in all pregnant rats with a peak value of 202 ng/ml on day 14. Elevated progesterone levels were associated negatively with LH and estradiol levels during pregnancy. The levels of estradiol surged significantly on day 21. Immunohistochemistry of the ovary showed low levels of OSE cells staining positive for ERα expression. ERα positive cells were absent on day 7 and 14 of pregnancy, only day 21 recorded a very low percentage of immunostaining (0.5%) within the nuclei of OSE cells. On the contrary, immunostaining of PR was not observed within the nuclei of OSE cells in all groups of study. In conclusions, these results may suggest that progesterone effect during pregnancy seems to be overriding the positive effect of estrogens on OSE cells. High progesterone levels may have a direct negative effect on gonadotropin production and thereby it might inhibit events leading to both follicular development and OSE proliferation. Understanding the factors affecting OSE proliferation may help elucidating the mechanism(s) of assisted diseases such as ovarian cancer.

Keywords: ovarian surface, pregnancy, gonadotropins, steroids

Procedia PDF Downloads 289

Authors: Sarim Ahmad

Abstract:

The Single Cell Gel Electrophoresis Assay (SCGE, known as comet assay) is a potential, uncomplicated, sensitive and state-of-the-art technique for quantitating DNA damage at individual cell level and repair from in vivo and in vitro samples of eukaryotic cells and some prokaryotic cells, being popular in its widespread use in various areas including human biomonitoring, genotoxicology, ecological monitoring and as a tool for research into DNA damage or repair in different cell types in response to a range of DNA damaging agents, cancer risk and therapy. The method involves the encapsulation of cells in a low-melting-point agarose suspension, lysis of the cells in neutral or alkaline (pH > 13) conditions, and electrophoresis of the suspended lysed cells, resulting in structures resembling comets as observed by fluorescence microscopy; the intensity of the comet tail relative to the head reflects the number of DNA breaks. The likely basis for this is that loops containing a break lose their supercoiling and become free to extend towards the anode. This is followed by visual analysis with staining of DNA and calculating fluorescence to determine the extent of DNA damage. This can be performed by manual scoring or automatically by imaging software. The assay can, therefore, predict an individual’s tumor sensitivity to radiation and various chemotherapeutic drugs and further assess the oxidative stress within tumors and to detect the extent of DNA damage in various cancerous and precancerous lesions of oral cavity.

Keywords: comet assay, single cell gel electrophoresis, DNA damage, early detection test

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Authors: Xiaoyan Jiang, Huizhu Wang, Lili Gui, Dandan Shen, Xiao Wei, Xi Yu, Hailiang Liu, Min Hong

Abstract:

Objective: Applicate FITC to establish Th2-type allergic contact dermatitis model, and study the effect and mechanism of Cimifugin on Th2-type allergic contact dermatitis. Methods: The Balb/c mice were sensitized with painting 80 ul of 1.5% FITC onto the shaved abdomen skin at DAY1 and DAY2. The animals were challenged on their right ears with 20 ul of 0.6% FITC, and the left ears were painted with solvent alone at day 6, mice were administered cimifugin for 7 days. 24h later, ear swelling was noted, and the infiltration of eosinophils was investigated by hematoxylin and eosin (H&E) staining. while part of the ear tissue homogenates prepared for detecting interleukin-4 levels by ELISA .Mice were administered cimifugin In the initial stage of the above model for 5 days(-1DAY—DAY3), ear tissue were homogenized to detect IL-33 levels by ELISA. Results: Cimifugin 25mg/kg, 50mg/kg inhibited mouse ear swelling, ear histopathology showed that mice given Cimifugin has significantly reduced levels of local tissue fluid exudation, congestion, infiltration of lymphocytes, and other inflammatory conditions compared with the model group. At the same time, it has significantly reduce of Th2 cytokines IL-4 in the mouse ear tissue homogenate. Data of the initial stage shows that 12.5mg/kg, 50mg/kg Cimifugin significantly inhibited IL-33 levels. Conclusion: Cimifugin inhibit FITC-induced Th2-type allergic contact dermatitis, and its mechanism may be related to inhibition of IL-33.

Keywords: cimifugin, allergic contact dermatitis, Th1/Th2, IL-33

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Authors: Manasa Perikala, Asha Bhardwaj

Abstract:

Fluorescent carbon dots (CDs), a young member of Carbon nanomaterial family, has gained a lot of research attention across the globe due to its highly luminescent emission properties, non-toxic behavior, stable emission properties, and zero re-absorption lose. These dots have the potential to replace the use of traditional semiconductor quantum dots in light-emitting devices (LED’s, fiber lasers) and other photonic devices (temperature sensor, UV detector). However, One major drawback of Carbon dots is that, till date, the actual mechanism of photoluminescence (PL) in carbon dots is still an open topic of discussion among various researchers across the globe. PL mechanism of CDs based on wide particle size distribution, the effect of surface groups, hybridization in carbon, and charge transfer mechanisms have been proposed. Although these mechanisms explain PL of CDs to an extent, no universally accepted mechanism to explain complete PL behavior of these dots is put forth. In our work, we report parameters affecting the size and surface of CDs, such as time of the reaction, synthesis temperature and concentration of precursors and their effects on the optical properties of the carbon dots. The effect of auxochromes on the emission properties and re-modification of carbon surface using an external surface functionalizing agent is discussed in detail. All the explanations have been supported by UV-Visible absorption, emission spectroscopies, Fourier transform infrared spectroscopy and Transmission electron microscopy and X-Ray diffraction techniques. Once the origin of PL in CDs is understood, parameters affecting PL centers can be modified to tailor the optical properties of these dots, which can enhance their applications in the fabrication of LED’s and other photonic devices out of these carbon dots.

Keywords: carbon dots, photoluminescence, size effects on emission in CDs, surface modification of carbon dots

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Authors: Shu-Jen Chen, Yi-Chien Wan, Ruey-Chi Wang

Abstract:

Hydroxyapatite (Ca10(PO4)6(OH)2, HAp) can be applied to the fabrication of bone replacement materials, the composite of dental filling, and the adsorption of biomolecules and dyes. The integration of HAp and magnetic materials would offer several advantages for bio-separation process because the magnetic adsorbents is capable of recovered by applied magnetic field. C-phycocyanin (C-PC) and Allophycocyanin (APC), isolated from Spirulina platensis, can be used in fluorescent labeling probes, health care foods and clinical diagnostic reagents. Although the purification of C-PC and APC are reported by HAp adsorption, the adsorption of C-PC and APC by magnetic HAp composites was not reported yet. Therefore, the fabrication of HAp with magnetic silica nanoparticles for proteins adsorption was investigated in this work. First, the magnetic silica particles were prepared by covering silica layer on Fe3O4 nanoparticles with a reverse micelle method. Then, the Fe3O4@SiO2 nanoparticles were mixed with calcium carbonate to obtain magnetic silica/calcium carbonate composites (Fe3O4@SiO2/CaCO3). The Fe3O4@SiO2/CaCO3 was further reacted with K2HPO4 for preparing the magnetic silica/hydroxyapatite composites (Fe3O4@SiO2/HAp). The adsorption experiments indicated that the adsorption capacity of Fe3O4@SiO2/HAp toward C-PC and APC were highest at pH 6. The adsorption of C-PC and APC by Fe3O4@SiO2/HAp could be correlated by the pseudo-second-order model, indicating chemical adsorption dominating the adsorption process. Furthermore, the adsorption data showed that the adsorption of Fe3O4@SiO2/HAp toward C-PC and APC followed the Langmuir isotherm. The isoelectric points of C-PC and APC were around 5.0. Additionally, the zeta potential data showed the Fe3O4@SiO2/HAp composite was negative charged at pH 6. Accordingly, the adsorption mechanism of Fe3O4@SiO2/HAp toward C-PC and APC should be governed by hydrogen bonding rather than electrostatic interaction. On the other hand, as compared to C-PC, the Fe3O4@SiO2/HAp shows higher adsorption affinity toward APC. Although the Fe3O4@SiO2/HAp cannot recover C-PC and APC from Spirulina platensis homogenate, the Fe3O4@SiO2/HAp can be applied to separate C-PC and APC.

Keywords: hydroxyapatite, magnetic, C-phycocyanin, allophycocyanin

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Authors: Indu Barwal, Shloka Thakur, Subhash C. Yadav

Abstract:

The plant virus capsid protein based nanoparticles are extensively studied for their application in biomedical research for development of nanomedicines and drug delivery systems. We have developed a chimeric drug delivery system by controlled in vitro assembly of separately bioconjugated fluorescent dye (as reporting molecule), folic acid (as receptor binding biomolecule for targeted delivery) and doxorubicin (as anticancer drug) using modified EDC NHS chemistry on heterologously overexpressed (E. coli) capsid proteins of cowpea chlorotic mottle virus (CCMV). This chimeric vehicle was further encapsidated on gold nanoparticles (20nm) coated with 5≠ thiolated DNA probe to neutralize the positive charge of capsid proteins. This facilitates the in vitro assembly of modified capsid subunits on the gold nanoparticles to develop chimeric GNPs encapsidated targeted drug delivery system. The bioconjugation of functionalities, number of functionality on capsid subunits as well as virus like nanoparticles, structural stability and in vitro assembly were confirmed by SDS PAGE, relative absorbance, MALDI TOF, ESI-MS, Circular dichroism, intrinsic tryptophan fluorescence, zeta particle size analyzer and TEM imaging. This vehicle was stable at pH 4.0 to 8.0 suitable for many organelles targeting. This in vitro assembled chimeric plant virus like particles could be suitable for ideal drug delivery vehicles for subcutaneous cancer treatment and could be further modified for other type of cancer treatment by conjugating other functionalities (targeting, drug) on capsids.

Keywords: chimeric drug delivery vehicles, bioconjugated plant, virus, capsid

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Authors: Sujata Chaturvedi, Ishita Pant, Deepak Kumar Jha, Vinod Kumar Singh Gautam, Chandra Bhushan Tripathi

Abstract:

Objective: To describe clinical, histopathological and immunohistochemical profile of glioblastoma in patients and to correlate these findings with patient survival. Material and methods: 30 cases of histopathologically diagnosed glioblastomas were included in this study. These cases were analysed in detail for certain clinical and histopathological parameters. Immunohistochemical staining for p53, epidermal growth factor receptor (EGFR), vascular endothelial growth factor (VEGF), mouse double minute 2 homolog (MDM2) and Ki67 was done and scores were calculated. Results of these findings were correlated with patient survival. Results: A retrospective analysis of the histopathology records and clinical case files was done in 30 cases of glioblastoma (WHO grade IV). The mean age of presentation was 50.6 years with a male predilection. The most common involved site was the frontal lobe. Amongst the clinical parameters, age of the patient and extent of surgical resection showed a significant correlation with the patient survival. Histopathological parameters showed no significant correlation with the patient survival, while amongst the immunohistochemical parameters expression of MDM2 showed a significant correlation with the patient survival. Conclusion: In this study incorporating clinical, histopathological and basic panel of immunohistochemistry, age of the patient, extent of the surgical resection and expression of MDM2 showed significant correlation with the patient survival.

Keywords: glioblastoma, p53, EGFR, VEGF, MDM2, Ki67

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Authors: Stefan Gruden, Charlott Brunmark, Bo Holmqvist, Erwin D. Brenndorfer, Martin Johansson, Jian Liu, Ying Zhao, Niklas Axen, Moustapha Hassan

Abstract:

Aim: The study was designed to evaluate the ability of the calcium sulfate-based NanoZolid® drug delivery technology to locally release the epidermal growth factor (EGF) protein while maintaining its biological activity. Methods: NanoZolid-formulated EGF protein labelled with a near-infrared dye (EGF-NIR) depots or EGF-NIR dissolved in PBS were injected subcutaneously into mice bearing EGF receptor (EGFR) positive human A549 lung cancer tumors inoculated subcutaneously. The release and biodistribution of the EGF-NIR were investigated in vivo longitudinally up to 96 hours post-administration, utilizing whole-body fluorescence imaging. In order to confirm the in vivo findings, histological analysis of tumor cryosections was performed to investigate EGF-NIR fluorescent signal and EGFR expression level by immunofluorescence labelling. Results: The in vivo fluorescence imaging showed a controlled release profile of the EGF-NIR loaded in the NanoZolid depots compared to free EGF-NIR. Histological analysis of the tumors further demonstrated a prevailing distribution of EGF-NIR in regions with high levels of EGFR expression. Conclusion: Calcium sulfate based depots can be used to formulate EGF while maintaining its biological activity, e.g., receptor binding capability. This may have good clinical potential for local delivery of biomolecules to enhance treatment efficacy and minimize systemic adverse effects.

Keywords: bioresorbable, calcium sulfate, controlled release, NanoZolid

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Authors: Faisal Nouroz, Mukaramin Mukaramin

Abstract:

Of the transposable elements (TEs), the retrotransposons are the most copious elements identified from many sequenced genomes. They have played a major role in genome evolution, rearrangement, and expansions based on their copy and paste mode of proliferation. They are further divided into LTR and Non-LTR retrotransposons. The purpose of the current study was to identify the LTR REs in sequenced Phoenix dactylifera genome and to study their structural diversity. A total of 150 P. dactylifera BAC sequences with > 60kb sizes were randomly retrieved from National Center for Biotechnology Information (NCBI) database and screened for the presence of LTR retrotransposons. Seven bacterial artificial chromosomes (BAC) sequences showed full-length LTR Retrotransposons with 4 Copia and 3 Gypsy families having variable copy numbers in respective families. Reverse transcriptase (RT) domain was found as the most conserved domain among Copia and Gypsy superfamilies and was used to deduce evolutionary analysis. The amino acid residues among various RT sequences showed variability in their percentages indicating post divergence evolution. Amino acid Leucine was found in highest proportions followed by Lysine, while Methionine and Tryptophan were in lowest percentages. The phylogenetic analysis based on RT domains confirmed that although having most conserved RT regions, several evolutionary events occurred causing nucleotide polymorphisms and hence clustering of Gypsy and Copia superfamilies into their respective lineages. The study will be helpful in identification and annotation of these elements in other species and genera and their distribution patterns on chromosomes by fluorescent in situ hybridization techniques.

Keywords: transposable elements, Phoenix dactylifera, retrotransposons, phylogenetic analysis

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Authors: R. M. Korzerzer, J. O. Hambolu, S. O. Salami, S. B. Oladele

Abstract:

The grasscutter (Thryonomys swinderianus) has become an important source of protein and income to rural dwellers in most West African countries including Nigeria. Twelve apparently healthy grasscutters consisting of six males and six females between the ages of three and seven months were obtained from rural dwellers in Benue state and used for this study. The animals were transported by means of constructed cages to the Department of Veterinary Anatomy, Ahmadu Bello University, Zaria and sacrificed using chloroform inhalation gaseous anaesthesia by suffocation. The spleen and mesenteric lymph nodes were extirpated and the tissues prepared using standard methods, haematoxilin and eosin stain was used for routine histology, while Rhodamine B-aniline-methylene blue stain was used for staining reticular and elastic fibres. The spleen was dark red in colour and roughly triangular in outline, and was observed to increase consistently with age, maximum values were recorded at seven months of age in both males and females. Mean ± SEM values for splenic weights were 0.67 ± 0.09 g, 1.65 ± 0.35 g and 2.31 ± 0.06 g at three, five and seven months of age, respectively. The percentage ratio of splenic weight to body weight was 0.1%. Histologically, the germinal centres revealed three zones; the germinal centre, cortical layer and the marginal zone. The mesenteric lymph nodes were constantly bean shaped and appeared as opaque white masses which resemble fat but were distinguished from fat by their pearly glossy nature. The mean ± SEM values for mesenteric lymph node weights were 0.056 ± 0.005 g, 0.143 ± 0.034 g and 0.1600 ± 0.023 g at three, five and seven months of age, respectively.

Keywords: anatomical, spleen, mesenteric lymph node, grasscutter

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