Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 530

Search results for: endothelial progenitor cells

530 Endothelial Specificity of ICAM2, Flt-1, and Tie2 Promoters In Vitro and In Vivo

Authors: Jing Lei, Yoram Vodovotz, Timothy R. Billiar

Abstract:

To identify an endothelial cell-specific promoter suitable for vascular-specific targeting, we tested five promoters in vitro--Tie2SE, Tie2LE, ICAM2, Flt-1 and vWF--for promoter activity and specificity in endothelial cells, smooth muscle cells and non-vascular resident cells as well as tissues. These promoters, except for vWF, exhibited good endothelial activity and specificity in vitro. In a syngenic heart transplantation model, the ICAM2 promoter was variably functional in coronary endothelial cells of donor hearts. Thus, the ICAM2, Flt-1, Tie2SE and Tie2LE promoters hold promise for endothelial-specific targeting, but in vitro expression may not predict in vivo expression.

Keywords: vascular-specific targeting, endothelial cell-specificpromoter, endothelial specificity.

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529 Effects of Stiffness on Endothelial Cells Behavior

Authors: Forough Ataollahi, Belinda Pingguan-Murphy, Wan Abu Bakar Wan Abas, Noor Azuan Abu Osman

Abstract:

Endothelium proliferation is an important process in cardiovascular homeostasis and can be regulated by extracellular environment, as cells can actively sense mechanical environment. In this study, we evaluated endothelial cell proliferation on PDMS/alumina (Al2O3) composites and pure PDMS. The substrates were prepared from pure PDMS and its composites with 5% and 10% Al2O3 at curing temperature 50˚C for 4h and then characterized by mechanical, structural and morphological analyses. Higher stiffness was found in the composites compared to the pure PDMS substrate. Cell proliferation of the cultured bovine aortic endothelial cells on substrate materials were evaluated via Resazurin assay and 1, 1’-Dioctadecyl-1, 3, 3, 3’, 3’-Tetramethylindocarbocyanine Perchlorate-Acetylated LDL (Dil-Ac-LDL) cell staining, respectively. The results revealed that stiffer substrates promote more endothelial cells proliferation to the less stiff substrates. Therefore, this study firmly hypothesizes that the stiffness elevates endothelial cells proliferation.

Keywords: Bovine aortic endothelial cells, extra cellular matrix, proliferation, stiffness.

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528 Endothelial-Cell-Mediated Displacement of Extracellular Matrix during Angiogenesis

Authors: Yue Du, Sahan C. B. Herath, Qing-Guo Wang, Harry Asada, Peter C. Y. Chen

Abstract:

Mechanical interaction between endothelial cells (ECs) and the extracellular matrix (or collagen gel) is known to influence the sprouting response of endothelial cells during angiogenesis. This influence is believed to impact on the capability of endothelial cells to sense soluble chemical cues. Quantitative analysis of endothelial-cell-mediated displacement of the collagen gel provides a means to explore this mechanical interaction. Existing analysis in this context is generally limited to 2D settings. In this paper, we investigate the mechanical interaction between endothelial cells and the extracellular matrix in terms of the endothelial-cellmediated displacement of the collagen gel in both 2D and 3D. Digital image correlation and Digital volume correlation are applied on confocal reflectance image stacks to analyze cell-mediated displacement of the gel. The skeleton of the sprout is extracted from phase contrast images and superimposed on the displacement field to further investigate the link between the development of the sprout and the displacement of the gel.

Keywords: Angiogenesis, digital image correlation, digital volume correlation, interaction between ECs and ECM.

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527 Biomolecules Based Microarray for Screening Human Endothelial Cells Behavior

Authors: Adel Dalilottojari, Bahman Delalat, Frances J. Harding, Michaelia P. Cockshell, Claudine S. Bonder, Nicolas H. Voelcker

Abstract:

Endothelial Progenitor Cell (EPC) based therapies continue to be of interest to treat ischemic events based on their proven role to promote blood vessel formation and thus tissue re-vascularisation. Current strategies for the production of clinical-grade EPCs requires the in vitro isolation of EPCs from peripheral blood followed by cell expansion to provide sufficient quantities EPCs for cell therapy. This study aims to examine the use of different biomolecules to significantly improve the current strategy of EPC capture and expansion on collagen type I (Col I). In this study, four different biomolecules were immobilised on a surface and then investigated for their capacity to support EPC capture and proliferation. First, a cell microarray platform was fabricated by coating a glass surface with epoxy functional allyl glycidyl ether plasma polymer (AGEpp) to mediate biomolecule binding. The four candidate biomolecules tested were Col I, collagen type II (Col II), collagen type IV (Col IV) and vascular endothelial growth factor A (VEGF-A), which were arrayed on the epoxy-functionalised surface using a non-contact printer. The surrounding area between the printed biomolecules was passivated with polyethylene glycol-bisamine (A-PEG) to prevent non-specific cell attachment. EPCs were seeded onto the microarray platform and cell numbers quantified after 1 h (to determine capture) and 72 h (to determine proliferation). All of the extracellular matrix (ECM) biomolecules printed demonstrated an ability to capture EPCs within 1 h of cell seeding with Col II exhibiting the highest level of attachment when compared to the other biomolecules. Interestingly, Col IV exhibited the highest increase in EPC expansion after 72 h when compared to Col I, Col II and VEGF-A. These results provide information for significant improvement in the capture and expansion of human EPC for further application.

Keywords: Cardiovascular disease, cell microarray platform, cell therapy, endothelial progenitor cells, high throughput screening.

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526 Evaluation of Antiglycation Effects of Extracts Obtained from Canarium album Raeusch Fruit and Beneficial Activity on Advanced Glycation Endproduct-Mediated Oxidative Stress and Inflammation in Monocytes and Vascular Endothelial Cells

Authors: Chiung-Tsun Kuoa, Tzu-Hao Liu, Fang-Yi Lin, Tai-Hao Hsu, Hui-Yin Chen

Abstract:

Hyperglycemia-mediated accumulation of advanced glycation end-products (AGEs) play a pivotal role in the development of diabetic complications by inducing inflammation. In the present study, we evaluated the possible effects of water/ethanol (1/1, v/v) extracts (WEE) and its fractions from Canarium album Raeusch. (Chinese olive) which is a fruit used on AGEs-stimulated oxidative stress and inflammation in monocytes and vascular endothelial cells. Co-incubation of EA.hy926 endothelial cells with WEE and its fractions for 24h resulted in a significant decrease of monocyte–endothelial cell adhesion, the expression of ICAM-1, generation of intracellular ROS and depletion of GSH induced by AGEs. Chinese olive fruit extracts also reduced the expression of pro-inflammatory mediates, such as TNF-α, IL-1β and IL-6 in THP-1 cells. These findings suggested that Chinese olive fruit was able to protect vascular endothelium from dysfunction induced by AGEs. 

Keywords: Advanced glycation end-products (AGEs), Canarium album Raeusch, endothelial dysfunction, inflammation.

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525 Potential Effects of Human Bone Marrow Non- Mesenchymal Mononuclear Cells on Neuronal Differentiation

Authors: Chareerut Phruksaniyom, Khwanthana Grataitong, Permphan Dharmasaroja, Surapol Issaragrisil

Abstract:

Bone marrow-derived stem cells have been widely studied as an alternative source of stem cells. Mesenchymal stem cells (MSCs) were mostly investigated and studies showed MSCs can promote neurogenesis. Little is known about the non-mesenchymal mononuclear cell fraction, which contains both hematopoietic and nonhematopoietic cells, including monocytes and endothelial progenitor cells. This study focused on unfractionated bone marrow mononuclear cells (BMMCs), which remained 72 h after MSCs were adhered to the culture plates. We showed that BMMC-conditioned medium promoted morphological changes of human SH-SY5Y neuroblastoma cells from an epithelial-like phenotype towards a neuron-like phenotype as indicated by an increase in neurite outgrowth, like those observed in retinoic acid (RA)-treated cells. The result could be explained by the effects of trophic factors released from BMMCs, as shown in the RT-PCR results that BMMCs expressed nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), and ciliary neurotrophic factor (CNTF). Similar results on the cell proliferation rate were also observed between RA-treated cells and cells cultured in BMMC-conditioned medium, suggesting that cells creased proliferating and differentiated into a neuronal phenotype. Using real-time RT-PCR, a significantly increased expression of tyrosine hydroxylase (TH) mRNA in SHSY5Y cells indicated that BMMC-conditioned medium induced catecholaminergic identities in differentiated SH-SY5Y cells.

Keywords: bone marrow, neuronal differentiation, neurite outgrowth, trophic factor, tyrosine hydroxylase

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524 Hemocompatible Thin-Film Materials Recreating the Structure of the Cell Niches with High Potential for Endothelialization

Authors: Roman Major, Klaudia Trembecka-Wojciga, Juergen Markus Lackner, Boguslaw Major

Abstract:

The future and the development of science is therefore seen in interdisciplinary areas such as biomedical engineering. Selfassembled structures, similar to stem cell niches would inhibit fast division process and subsequently capture the stem cells from the blood flow. By means of surface topography and the stiffness as well as microstructure progenitor cells should be differentiated towards the formation of endothelial cells monolayer which effectively will inhibit activation of the coagulation cascade. The idea of the material surface development met the interest of the clinical institutions, which support the development of science in this area and are waiting for scientific solutions that could contribute to the development of heart assist systems. This would improve the efficiency of the treatment of patients with myocardial failure, supported with artificial heart assist systems. Innovative materials would enable the redesign, in the post project activity, construction of ventricular heart assist.

Keywords: Bio-inspired materials, electron microscopy, haemocompatibility, niche-like structures, thin coatings.

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523 On the Mathematical Model of Vascular Endothelial Growth Connected with a Tumor Proliferation

Authors: N. Khatiashvili, Ch. Pirumova, V. Akhobadze

Abstract:

In the paper the mathematical model of tumor growth is considered. New capillary network formation, which supply cancer cells with the nutrients, is taken into the account. A formula estimating a tumor growth in connection with the number of capillaries is obtained.

Keywords: Differential Equations, Mathematical Models, Vascular Endothelial, Tumor

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522 High Glucose Increases Acetylcholine-Induced Ca2+ Entry and Protein Expression of STIM1

Authors: Hong Ding, Fatiha Benslimane, Isra Marei, Chris R. Triggle

Abstract:

Hyperglycaemia is a key factor that contributes to the development of diabetes-related microvascular disease and a major risk factor for endothelial dysfunction. In the current study, we have explored glucose-induced abnormal intracellular calcium (Ca2+ i) homeostasis in mouse microvessel endothelial cells (MMECs) in high glucose (HG) (40mmol/L) versus control (low glucose, LG) (11 mmol/L). We demonstrated that the exposure of MMECs to HG for 3 days did not change basal Ca2+ i, however, there was a significant increase of acetylcholine-induced Ca2+ entry. Western blots illustrated that exposure to HG also increased STIM1 (Stromal Interaction Molecule 1), but not Orai1 (the pore forming subunit), protein expression levels. Although the link between HG-induced changes in STIM1 expression, enhanced Ca2+ entry and endothelial dysfunction requires further study, the current data are suggestive that targeting these pathways may reduce the impact of HG on endothelial function.

Keywords: store-operated calcium entry, hyperglycaemia, STIM1, endothelial dysfunction

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521 Stroma-Providing Activity of Adipose Derived Mesenchymal Stromal Cells in Tissue-Related O2 Microenvironment

Authors: P. I. Bobyleva, E. R. Andreeva, I. V. Andrianova, E. V. Maslova, L.B. Buravkova

Abstract:

This work studied the ability of adipose tissue-derived mesenchymal stromal cells (MSCs) to form stroma for expansion of cord blood hematopoietic cells. We showed that 72-hour interaction of MSCs with cord blood mononuclear cells (MNCs) in vitro at atmospheric (20%) and low (5%) O2 conditions increased the expression of ICAM-1, HCAM (at the beginning of interaction) on MSCs. Viability of MSCs and MNCs were maintained at high level. Adhesion of MNCs to MSCs was faster at 20% O2. MSCs promoted the proliferation of adhered MNCs to form the suspension containing great number of hematopoietic colony-forming units, and this effect was more pronounced at 5% O2. Thus, adipose-derived MSCs supplied sufficient stromal support to cord blood MNCs both at 20% and 5% О2, providing their adhesion with further expansion of new generation of different hematopoietic lineages.

Keywords: Hematopoietic stem and progenitor cells, mesenchymal stromal cells, tissue-related oxygen.

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520 Tumor Necrosis Factor-α Regulates Heme Oxygenase-1 Expression in Endothelial Cells via the Phosphorylation of JNK/p38

Authors: Chan-Jung Liang, Shu-Huei Wang, Pei-Jhen Wu, Jaw-Shiun Tsai, Chau-Chung Wu, Yuh-Lien Chen

Abstract:

Heme oxygenase-1 (HO-1), an enzyme degrading heme to carbon monoxide, iron, and biliverdin, has been recognized as playing a crucial role in cellular defense against stressful conditions, not only related to heme release. In the present study, the effects of TNF-a on the expression of heme oxygenase-1 (HO-1) in human aortic endothelial cells (HAECs) as well as the related mechanisms were investigated. 10 ng/mL TNF-α treatment significantly increased HO-1 expression after 6h, then a further increase at 12h and declined at 24h. Treatment with 2 ng/mL of TNF-a after 12 h resulted in a significant increase in HO-1 expression, which peaked at 10 ng/mL, then declined at 20 ng/mL. TNF-α induced HO-1 expression and then HO-1 expression reduced  vascular cell adhesion molecule-1 (VCAM-1) expression. Phosphorylation studies of ERK1/2, JNK, and p38, three subgroups of mitogen-activated protein kinases (MAPKs) demonstrated TNF-α-induced ERK1/2, JNK, and p38 phosphorylation. The increase in HO-1 expression in response to TNF-α treatment was affected by pretreatment with SP600125 (a JNK inhibitor) and SB203580 (a p38 inhibitor), not with PD98059 (an ERK1/2 inhibitor). The expression of HO-1 was stronger in aortas of TNF-α-treated apo-E deficient mice when compared with control mice. These results suggest that low dose of TNF-α treatment notably induced HO-1 expression was mediated through JNK/p38 phosphorylation and may have a protective potential in cardiovascular diseases and inflammatory response through the regulation of HO-1 expression.

Keywords: Heme oxygenase-1 inflammation, endothelial cells, mitogen-activated protein kinases (MAPKs).

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519 Effect of Copper Ions Doped-Hydroxyapatite 3D Fiber Scaffold

Authors: Adil Elrayah, Jie Weng, Esra Suliman

Abstract:

The mineral in human bone is not pure stoichiometric calcium phosphate (Ca/P) as it is partially substituted by in organic elements. In this study, the copper ions (Cu2+) substituted hydroxyapatite (CuHA) powder has been synthesized by the co-precipitation method. The CuHA powder has been used to fabricate CuHA fiber scaffolds by sol-gel process and the following sinter process. The resulted CuHA fibers have slightly different microstructure (i.e. porosity) compared to HA fiber scaffold, which is denser. The mechanical properties test was used to evaluate CuHA, and the results showed decreases in both compression strength and hardness tests. Moreover, the in vitro used endothelial cells to evaluate the angiogenesis of CuHA. The result illustrated that the viability of endothelial cell on CuHA fiber scaffold surfaces tends to antigenic behavior. The results obtained with CuHA scaffold give this material benefit in biological applications such as antimicrobial, antitumor, antigens, compacts, filling cavities of the tooth and for the deposition of metal implants anti-tumor, anti-cancer, bone filler, and scaffold.

Keywords: Fiber scaffold, copper ions, hydroxyapatite, hardness, in vitro, mechanical properties.

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518 Oleate Induces Apoptosis in 3T3-L1 Adipocytes

Authors: A. Rohana, A. M., Fadzilah Adibah, M. S. Muhammad Roji

Abstract:

Oleic acid (C18:1) play an important role in proliferation of fat cells. In this study, the effect of oleate on cells viability in 3T3-L1 cells (fat cells) was investigated. The 3T3-L1 cells were treated with various concentrations of oleate in the presence of 23 mM glucose. Oleate was added to adipogenic media (day 0) to investigate the influence of oleate on proliferation of postconfluent preadipocytes after 24 h induction. 0.1 mM oleate promoted cell division by increasing 33.9% number of cells from basal control in postconfluent preadipocytes. However, there were no significantly different in cells viability with control cells when oleate concentrations were increased up to 0.5 mM. When added to differentiated adipocytes (day 12) for 48 h, the number of cells decreased as oleate concentrations increased. 92.7% of cells lost demonstrated apoptosis and necrosis after 48 h with 0.5 mM oleate. The fluorochrome staining was examined under fluorescence microscopy using acridine orange and ethidium bromide double staining. Furthermore, the presence of high lactate (60.6% increased from basal control) released into plasma has shown the direct cytotoxicity of 0.5 mM oleate on adipocytes.

Keywords: adipocytes, apoptosis, oleate, postconfluentpreadipocytes

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517 Comparison of the Performance of GaInAsSb and GaSb Cells under Different Temperature Blackbody Radiations

Authors: Liangliang Tang, Chang Xu, Xingying Chen

Abstract:

GaInAsSb cells probably show better performance than GaSb cells in low-temperature thermophotovoltaic systems due to lower bandgap; however, few experiments proved this phenomenon so far. In this paper, numerical simulation is used to evaluate GaInAsSb and GaSb cells with similar structures under different radiation temperatures. We found that GaInAsSb cells with n-type emitters show slightly higher output power densities compared with that of GaSb cells with n-type emitters below 1,550 K-blackbody radiation, and the power density of the later cells will suppress the formers above this temperature point. During the temperature range of 1,000~2,000 K, the efficiencies of GaSb cells are about twice of GaInAsSb cells if perfect filters are used to prevent the emission of the non-absorbed long wavelength photons. Several parameters that affect the GaInAsSb cell were analyzed, such as doping profiles, thicknesses of GaInAsSb epitaxial layer and surface recombination velocity. The non-p junctions, i.e., n-type emitters are better for GaInAsSb cell fabrication, which is similar to that of GaSb cells.

Keywords: Thermophotovoltaic cell, GaSb, GaInAsSb, diffused emitters.

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516 Gene Expressions Associated with Ultrastructural Changes in Vascular Endothelium of Atherosclerotic Lesion

Authors: M. Maimunah, G.A. Froemming, H. Nawawi, M.I. Nafeeza, O. Effat, M.R. Rohayu Izanwati, M.S. Mohamed Saifulaman

Abstract:

Attachment of the circulating monocytes to the endothelium is the earliest detectable events during formation of atherosclerosis. The adhesion molecules, chemokines and matrix proteases genes were identified to be expressed in atherogenesis. Expressions of these genes may influence structural integrity of the luminal endothelium. The aim of this study is to relate changes in the ultrastructural morphology of the aortic luminal surface and gene expressions of the endothelial surface, chemokine and MMP-12 in normal and hypercholesterolemic rabbits. Luminal endothelial surface from rabbit aortic tissue was examined by scanning electron microscopy (SEM) using low vacuum mode to ascertain ultrastructural changes in development of atherosclerotic lesion. Gene expression of adhesion molecules, MCP-1 and MMP-12 were studied by Real-time PCR. Ultrastructural observations of the aortic luminal surface exhibited changes from normal regular smooth intact endothelium to irregular luminal surface including marked globular appearance and ruptures of the membrane layer. Real-time PCR demonstrated differentially expressed of studied genes in atherosclerotic tissues. The appearance of ultrastructural changes in aortic tissue of hypercholesterolemic rabbits is suggested to have relation with underlying changes of endothelial surface molecules, chemokine and MMP-12 gene expressions.

Keywords: Ultrastructure of luminal endothelial surface, Macrophage metalloelastase (MMP-12), Real-time PCR.

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515 The Identification of Anuran Glial Cells

Authors: Ibrahim M. S. Shnawa

Abstract:

Attempts were made to identify anuran glial cells. They were found as nervous tissue resident. Having stage dependent morphotype changes, whereby, appeared as an ovoid to oval in resting state and amoeboid mrophotypes in activated state, stained fairly with methylene blue and take up Pelikane blue 10% aqueous solution, as well as having the ability to phagocytize heat killed Staphylococcus aureus. They were delineated from the migrating peripheral monocytes by morphotypic and morphometeric differences. Such criteria were consistence with glial cells. Thus, the anuran glial cells are being identified in the frog Rana ridibunda Pallas 1771 and this animal can be of use as a simple model for the immunobiology of glial cells.

Keywords: Amoeboid cell, bacterial phagocytosis, Glial cells, Resting.

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514 Identification of Differentially Expressed Gene(DEG) in Atherosclerotic Lesion by Annealing Control Primer (ACP)-Based Genefishing™ PCR

Authors: M. Maimunah, G. A. Froemming, H. Nawawi, M. I. Nafeeza, O. Effat, M. Y. Rosmadi, M. S. Mohamed Saifulaman

Abstract:

Atherosclerosis was identified as a chronic inflammatory process resulting from interactions between plasma lipoproteins, cellular components (monocyte, macrophages, T lymphocytes, endothelial cells and smooth muscle cells) and the extracellular matrix of the arterial wall. Several types of genes were known to express during formation of atherosclerosis. This study is carried out to identify unknown differentially expressed gene (DEG) in atherogenesis. Rabbit’s aorta tissues were stained by H&E for histomorphology. GeneFishing™ PCR analysis was performed from total RNA extracted from the aorta tissues. The DNA fragment from DEG was cloned, sequenced and validated by Real-time PCR. Histomorphology showed intimal thickening in the aorta. DEG detected from ACP-41 was identified as cathepsin B gene and showed upregulation at week-8 and week-12 of atherogenesis. Therefore, ACP-based GeneFishing™ PCR facilitated identification of cathepsin B gene which was differentially expressed during development of atherosclerosis.

Keywords: Atherosclerosis, GeneFishing™ PCR, cathepsin B gene.

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513 Enhanced Differentiation of Stromal Cells and Embryonic Stem Cells with Vitamin D3

Authors: Mayada Alqaisi, Nasser Al-Shanti, Quiyu Wang, William S. Gilmore

Abstract:

In-vitro mouse co-culture of E14 embryonic stem cells (ESCs) and OP9 stromal cells can recapitulate the earliest stages of haematopoietic development, not accessible in human embryos, supporting both haemogenic precursors and their primitive haematopoietic progeny. 1α, 25-Dihydroxy-vitamin D3 (VD3) has been demonstrated to be a powerful differentiation inducer for a wide variety of neoplastic cells, and could enhance early differentiation of ESCs into blood cells in E14/OP9 co-culture. This study aims to ascertain whether VD3 is key in promoting differentiation and suppressing proliferation, by separately investigating the effects of VD3 on the proliferation phase of the E14 cell line and on stromal OP9 cells.The results showed that VD3 inhibited the proliferation of the cells in a dose-dependent manner, quantitatively by decreased cell number, and qualitatively by alkaline-phosphatase staining that revealed significant differences between VD3-treated and untreated cells, characterised by decreased enzyme expression (colourless cells). Propidium-iodide cell-cycle analyses showed no significant percentage change in VD3-treated E14 and OP9 cells within their G and S-phases, compared to the untreated controls, despite the increased percentage of G-phase compared to the S-phase in a dosedependent manner. These results with E14 and OP9 cells indicate that adequate VD3 concentration enhances cellular differentiation and inhibits proliferation. The results also suggest that if E14 and OP9 cells were co-cultured andVD3-treated, there would be furtherenhanced differentiation of ESCs into blood cells.

Keywords: Differentiation, embryonic stem cells, OP9 stromal cells, , 25-dihydroxy-vitamin D3

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512 Effect of Oxytocin on Cytosolic Calcium Concentration of Alpha and Beta Cells in Pancreas

Authors: Rauza Sukma Rita, Katsuya Dezaki, Yuko Maejima, Toshihiko Yada

Abstract:

Oxytocin is a nine-amino acid peptide synthesized in the paraventricular nucleus (PVN) and supraoptic nucleus (SON) of the hypothalamus. Oxytocin promotes contraction of the uterus during birth and milk ejection during breast feeding. Although oxytocin receptors are found predominantly in the breasts and uterus of females, many tissues and organs express oxytocin receptors, including the pituitary, heart, kidney, thymus, vascular endothelium, adipocytes, osteoblasts, adrenal gland, pancreatic islets, and many cell lines. On the other hand, in pancreatic islets, oxytocin receptors are expressed in both α-cells and β-cells with stronger expression in α- cells. However, to our knowledge there are no reports yet about the effect of oxytocin on cytosolic calcium reaction on α and β-cell. This study aims to investigate the effect of oxytocin on α-cells and β-cells and its oscillation pattern. Islet of Langerhans from wild type mice were isolated by collagenase digestion. Isolated and dissociated single cells either α-cells or β-cells on coverslips were mounted in an open chamber and superfused in HKRB. Cytosolic concentration ([Ca2+]i) in single cells were measured by fura-2 microfluorimetry. After measurement of [Ca2+]i, α-cells were identified by subsequent immunocytochemical staining using an anti-glucagon antiserum. In β-cells, the [Ca2+]i increase in response to oxytocin was observed only under 8.3 mM glucose condition, whereas in α-cells, [Ca2+]i an increase induced by oxytocin was observed in both 2.8 mM and 8.3 mM glucose. The oscillation incidence was induced more frequently in β-cells compared to α-cells. In conclusion, the present study demonstrated that oxytocin directly interacts with both α-cells and β-cells and induces increase of [Ca2+]i and its specific patterns.

Keywords: α-cells, β-cells, cytosolic calcium concentration, oscillation, oxytocin.

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511 White Blood Cells Identification and Counting from Microscopic Blood Image

Authors: Lorenzo Putzu, Cecilia Di Ruberto

Abstract:

The counting and analysis of blood cells allows the evaluation and diagnosis of a vast number of diseases. In particular, the analysis of white blood cells (WBCs) is a topic of great interest to hematologists. Nowadays the morphological analysis of blood cells is performed manually by skilled operators. This involves numerous drawbacks, such as slowness of the analysis and a nonstandard accuracy, dependent on the operator skills. In literature there are only few examples of automated systems in order to analyze the white blood cells, most of which only partial. This paper presents a complete and fully automatic method for white blood cells identification from microscopic images. The proposed method firstly individuates white blood cells from which, subsequently, nucleus and cytoplasm are extracted. The whole work has been developed using MATLAB environment, in particular the Image Processing Toolbox.

Keywords: Automatic detection, Biomedical image processing, Segmentation, White blood cell analysis.

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510 Optimization of Breast Tumor Cells Isolation Efficiency and Purity by Membrane Filtration

Authors: Bhuvanendran Nair Gourikutty Sajay, Liu Yuxin, Chang Chia-Pin, Poenar Daniel Puiu, Abdur Rub Abdur Rahman

Abstract:

Size based filtration is one of the common methods employed to isolate circulating tumor cells (CTCs) from whole blood. It is well known that this method suffers from isolation efficiency to purity tradeoff. However, this tradeoff is poorly understood. In this paper, we present the design and manufacturing of a special rectangular slit filter. The filter was designed to retain maximal amounts of nucleated cells, while minimizing the pressure on cells, thereby preserving their morphology. The key parameter, namely, input pressure, was optimized to retain the maximal number of tumor cells, whilst maximizing the depletion of normal blood cells (red and white blood cells and platelets). Our results indicate that for a slit geometry of 5 × 40 μm on a 13 mm circular membrane with a fill factor of 21%, a pressure of 6.9 mBar yields the optimum for maximizing isolation of MCF-7 and depletion of normal blood cells.

Keywords: Circulating tumor cells, Parylene slit membrane, Retention, White Blood Cell depletion.

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509 Cellular Components of the Hemal Node of Egyptian Cattle

Authors: Amira E. Derbalah, Doaa M. Zaghloul

Abstract:

10 clinically healthy hemal nodes were collected from male bulls aged 2-3 years. Light microscopy revealed a capsule of connective tissue consisted mainly of collagen fiber surrounding hemal node, numerous erythrocytes were found in wide subcapsular sinus under the capsule. The parenchyma of the hemal node was divided into cortex and medulla. Diffused lymphocytes, and lymphoid follicles, having germinal centers were the main components of the cortex, while in the medulla there was wide medullary sinus, diffused lymphocytes and few lymphoid nodules. The area occupied with lymph nodules was larger than that occupied with non-nodular structure of lymphoid cords and blood sinusoids. Electron microscopy revealed the cellular components of hemal node including elements of circulating erythrocytes intermingled with lymphocytes, plasma cells, mast cells, reticular cells, macrophages, megakaryocytes and endothelial cells lining the blood sinuses. The lymphocytes were somewhat triangular in shape with cytoplasmic processes extending between adjacent erythrocytes. Nuclei were triangular to oval in shape, lightly stained with clear nuclear membrane indentation and clear nucleoli. The reticular cells were elongated in shape with cytoplasmic processes extending between adjacent lymphocytes, rough endoplasmic reticulum, ribosomes and few lysosomes were seen in their cytoplasm. Nucleus was elongated in shape with less condensed chromatin. Plasma cells were oval to irregular in shape with numerous dilated rough endoplasmic reticulum containing electron lucent material occupying the whole cytoplasm and few mitochondria were found. Nuclei were centrally located and oval in shape with heterochromatin emarginated and often clumped near the nuclear membrane. Occasionally megakaryocytes and mast cells were seen among lymphocytes. Megakaryocytes had multilobulated nucleus and free ribosomes often appearing as small aggregates in their cytoplasm, while mast cell had their characteristic electron dense granule in the cytoplasm, few electron lucent granules were found also, we conclude that, the main function of the hemal node of cattle is proliferation of lymphocytes. No role for plasma cell in erythrophagocytosis could be suggested.

Keywords: Cattle, Electron microscopy, Hemal node, Histology, Immune system.

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508 Microalbuminuria in Human Immunodeficiency Virus Infection and Acquired Immunodeficiency Syndrome

Authors: Sharan Badiger, Prema T. Akkasaligar, Patil LS, Manish Patel, Biradar MS

Abstract:

Human immunodeficiency virus infection and acquired immunodeficiency syndrome is a global pandemic with cases reporting from virtually every country and continues to be a common infection in developing country like India. Microalbuminuria is a manifestation of human immunodeficiency virus associated nephropathy. Therefore, microalbuminuria may be an early marker of human immunodeficiency virus associated nephropathy, and screening for its presence may be beneficial. A strikingly high prevalence of microalbuminuria among human immunodeficiency virus infected patients has been described in various studies. Risk factors for clinically significant proteinuria include African - American race, higher human immunodeficiency virus ribonucleic acid level and lower CD4 lymphocyte count. The cardiovascular risk factors of increased systolic blood pressure and increase fasting blood sugar level are strongly associated with microalbuminuria in human immunodeficiency virus patient. These results suggest that microalbuminuria may be a sign of current endothelial dysfunction and micro-vascular disease and there is substantial risk of future cardiovascular disease events. Positive contributing factors include early kidney disease such as human immunodeficiency virus associated nephropathy, a marker of end organ damage related to co morbidities of diabetes or hypertension, or more diffuse endothelial cells dysfunction. Nevertheless after adjustment for non human immunodeficiency virus factors, human immunodeficiency virus itself is a major risk factor. The presence of human immunodeficiency virus infection is independent risk to develop microalbuminuria in human immunodeficiency virus patient. Cardiovascular risk factors appeared to be stronger predictors of microalbuminuria than markers of human immunodeficiency virus severity person with human immunodeficiency virus infection and microalbuminuria therefore appear to potentially bear the burden of two separate damage related to known vascular end organ damage related to know vascular risk factors, and human immunodeficiency virus specific processes such as the direct viral infection of kidney cells.The higher prevalence of microalbuminuria among the human immunodeficiency virus infected could be harbinger of future increased risks of both kidney and cardiovascular disease. Further study defining the prognostic significance of microalbuminuria among human immunodeficiency virus infected persons will be essential. Microalbuminuria seems to be a predictor of cardiovascular disease in diabetic and non diabetic subjects, hence it can also be used for early detection of micro vascular disease in human immunodeficiency virus positive patients, thus can help to diagnose the disease at the earliest.

Keywords: Acquired immunodeficiency syndrome, Human immunodeficiency virus, Microalbuminuria.

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507 Different Formula of Mixed Bacteria as a Bio-Treatment for Sewage Wastewater

Authors: E. Marei, A. Hammad, S. Ismail, A. El-Gindy

Abstract:

This study aims to investigate the ability of different formula of mixed bacteria as a biological treatments of wastewater after primary treatment as a bio-treatment and bio-removal and bio-adsorbent of different heavy metals in natural circumstances. The wastewater was collected from Sarpium forest site-Ismailia Governorate, Egypt. These treatments were mixture of free cells and mixture of immobilized cells of different bacteria. These different formulas of mixed bacteria were prepared under Lab. condition. The obtained data indicated that, as a result of wastewater bio-treatment, the removal rate was found to be 76.92 and 76.70% for biological oxygen demand, 79.78 and 71.07% for chemical oxygen demand, 32.45 and 36.84 % for ammonia nitrogen as well as 91.67 and 50.0% for phosphate after 24 and 28 hrs with mixed free cells and mixed immobilized cells, respectively. Moreover, the bio-removals of different heavy metals were found to reach 90.0 and 50. 0% for Cu ion, 98.0 and 98.5% for Fe ion, 97.0 and 99.3% for Mn ion, 90.0 and 90.0% Pb, 80.0% and 75.0% for Zn ion after 24 and 28 hrs with mixed free cells and mixed immobilized cells, respectively. The results indicated that 13.86 and 17.43% of removal efficiency and reduction of total dissolved solids were achieved after 24 and 28 hrs with mixed free cells and mixed immobilized cells, respectively.

Keywords: Biological desalination, bio-sorption heavy metals, free cell bacteria, immobilized bacteria, wastewater bio-treatment.

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506 Temperature Effect on the Organic Solar Cells Parameters

Authors: F.Belhocine-Nemmar; MS.Belkaid D. Hatem, O Boughias

Abstract:

In this work, the influence of temperature on the different parameters of solar cells based on organic semiconductors are studied. The short circuit current Isc increases so monotonous with temperature and then saturates to a maximum value before decreasing at high temperatures. The open circuit voltage Vco decreases linearly with temperature. The fill factor FF and efficiency, which are directly related with Isc and Vco follow the variations of the letters. The phenomena are explained by the behaviour of the mobility which is a temperature activated process.

Keywords: cells parameters, organic materials, solar cells, temperature effect

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505 Immunomodulatory Effects of Multipotent Mesenchymal Stromal Cells on T-Cell Populations at Tissue-Related Oxygen Level

Authors: A. N. Gornostaeva, P. I. Bobyleva, E. R. Andreeva, L. B. Buravkova

Abstract:

Multipotent mesenchymal stromal cells (MSCs) possess immunomodulatory properties. The effect of MSCs on the crucial cellular immunity compartment – T-cells is of a special interest. It is known that MSC tissue niche and expected milieu of their interaction with T- cells are characterized by low oxygen concentration, whereas the in vitro experiments usually are carried out at a much higher ambient oxygen (20%). We firstly evaluated immunomodulatory effects of MSCs on T-cells at tissue-related oxygen (5%) after interaction implied cell-to-cell contacts and paracrine factors only. It turned out that MSCs under reduced oxygen can effectively suppress the activation and proliferation of PHAstimulated T-cells and can provoke decrease in the production of proinflammatory and increase in anti-inflammatory cytokines. In hypoxia some effects were amplified (inhibition of proliferation, antiinflammatory cytokine profile shift). This impact was more evident after direct cell-to-cell interaction; lack of intercellular contacts could revoke the potentiating effect of hypoxia.

Keywords: Cell-to-cell interaction, low oxygen, MSC immunosuppression, T-cells.

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504 Enhancing K-Means Algorithm with Initial Cluster Centers Derived from Data Partitioning along the Data Axis with the Highest Variance

Authors: S. Deelers, S. Auwatanamongkol

Abstract:

In this paper, we propose an algorithm to compute initial cluster centers for K-means clustering. Data in a cell is partitioned using a cutting plane that divides cell in two smaller cells. The plane is perpendicular to the data axis with the highest variance and is designed to reduce the sum squared errors of the two cells as much as possible, while at the same time keep the two cells far apart as possible. Cells are partitioned one at a time until the number of cells equals to the predefined number of clusters, K. The centers of the K cells become the initial cluster centers for K-means. The experimental results suggest that the proposed algorithm is effective, converge to better clustering results than those of the random initialization method. The research also indicated the proposed algorithm would greatly improve the likelihood of every cluster containing some data in it.

Keywords: Clustering algorithm, K-means algorithm, Datapartitioning, Initial cluster centers.

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503 Synchrony between Genetic Repressilators in Sister Cells in Different Temperatures

Authors: Jerome G. Chandraseelan, Samuel M. D. Oliveira, Antti Häkkinen, Sofia Startceva, Andre S. Ribeiro

Abstract:

We used live E. coli containing synthetic genetic oscillators to study how the degree of synchrony between the genetic circuits of sister cells changes with temperature. We found that both the mean and the variability of the degree of synchrony between the fluorescence signals from sister cells are affected by temperature. Also, while most pairs of sister cells were found to be highly synchronous in each condition, the number of asynchronous pairs increased with increasing temperature, which was found to be due to disruptions in the oscillations. Finally we provide evidence that these disruptions tend to affect multiple generations as opposed to individual cells. These findings provide insight in how to design more robust synthetic circuits and in how cell division can affect their dynamics.

Keywords: Repressilator, robustness, synchrony, synthetic biology.

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502 A Compilation of Nanotechnology in Thin Film Solar Cell Devices

Authors: Nurul Amziah Md Yunus, Izhal Abdul Halin, Nasri Sulaiman, Noor Faezah Ismail, Nik Hasniza Nik Aman

Abstract:

Nanotechnology has become the world attention in various applications including the solar cells devices due to the uniqueness and benefits of achieving low cost and better performances of devices. Recently, thin film solar cells such as Cadmium Telluride (CdTe), Copper-Indium-Gallium-diSelenide (CIGS), Copper-Zinc-Tin-Sulphide (CZTS), and Dye-Sensitized Solar Cells (DSSC) enhanced by nanotechnology have attracted much attention. Thus, a compilation of nanotechnology devices giving the progress in the solar cells has been presented. It is much related to nanoparticles or nanocrystallines, carbon nanotubes, and nanowires or nanorods structures.

Keywords: Nanotechnology, nanocrystalline, nanowires, carbon nanotubes, nanorods, thin film solar cells.

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501 Possible Role of Polyamine on Tumor Spread after Surgical Trauma

Authors: Kuniyasu Soda

Abstract:

Surgical trauma seems to facilitate metastatic spread, although the underlying mechanisms are not known. Increased concentrations of polyamines (spermine and spermidine) in the blood seem to have associated with the enhanced malignant potential of cancer cells and decrease in anti-tumor immunity of cancer patients. In addition to de novo synthesis in rapidly growing cells such as normal regenerating cells and cancer cells, cells can take up polyamines from extra-cellular sources. We have shown that increased polyamine concentration results in decreases in cytokine production and expression of adhesion molecules involved in anti-tumor immunity, such as CD11a. And, immune cells in an environment with increased polyamine levels lose anti-tumor immune functions, such as lymphokine activated killer cell (LAK) activities. Because blood polyamine levels are increased in post-surgical patients, polyamine seems to have roles on post-traumatic tumor spread.

Keywords: Immune function, LAK, Polyamine, Surgical trauma.

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