Search results for: HT-29 human epithelium cells.

Authors: Ali Q. S., Farid A. J., Kabeir B. M., Zamberi S., Shuhaimi M., Ghazali H. M., Yazid A. M.

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Adhesion to the human intestinal cell is considered as one of the main selection criteria of lactic acid bacteria for probiotic use. The adhesion ability of two Bifidobacteriums strains Bifidobacterium longum BB536 and Bifidobacterium psudocatenulatum G4 was done using HT-29 human epithelium cell line as in vitro study. Four different level of pH were used 5.6, 5.7, 6.6, and 6.8 with four different times 15, 30, 60, and 120 min. Adhesion was quantified by counting the adhering bacteria after Gram staining. The adhesion of B. longum BB536 was higher than B. psudocatenulatum G4. Both species showed significant different in the adhesion properties at the factors tested. The highest adhesion for both Bifidobacterium was observed at 120 min and the low adhesion was in 15 min. The findings of this study will contribute to the introduction of new effective probiotic strain for future utilization.

Keywords: Bifidobacterium, Adhesion, HT-29 human epithelium cells.

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Authors: Siti Aishah Abu Bakar, Madihah Zawawi, Abdul Manaf Ali, Aini Ideris

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Newcastle Disease Virus (NDV), an avian paramyxovirus, is a highly contagious, generalised virus disease of domestic poultry and wild birds characterized by gastro-intestinal, respiratory and nervous signs. In this study, it was shown that NDV strain AF2240 and V4-UPM are cytolytic to Human Promyelocytic Leukemia, HL60 and Human T-lymphoblastic Leukemia, CEM-SS cells. Results from MTT cytolytic assay showed that CD50 for NDV AF2240 against HL60 was 130 HAU and NDV V4-UPM against HL60 and CEM-SS were 110.6 and 150.9 HAU respectively. Besides, both strains were found to inhibit the proliferation of cells in a dose dependent manner. The mode of cell death either by apoptosis or necrosis was further analyzed using acridine orange and propidium iodide (AO/PI) staining. Our results showed that both NDV strains induced primarily apoptosis in treated cells at CD50 concentration. In conclusion, both NDV strains caused cytolytic effects primarily via apoptosis in leukemia cells.

Keywords: Apoptosis, Cytolytic, Leukaemia, Newcastle DiseaseVirus

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Authors: Ali A. Alshatwi, Vaiyapuri S. Periasamy, Jegan Athinarayanan

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Engineered nanoparticles’ usage rapidly increased in various applications in the last decade due to their unusual properties. However, there is an ever increasing concern to understand their toxicological effect in human health. Particularly, metal and metal oxide nanoparticles have been used in various sectors including biomedical, food and agriculture. But their impact on human health is yet to be fully understood. In this present investigation, we assessed the toxic effect of engineered nanoparticles (ENPs) including Ag, MgO and Co3O4 nanoparticles (NPs) on human mesenchymal stem cells (hMSC) adopting cell viability and cellular morphological changes as tools The results suggested that silver NPs are more toxic than MgO and Co3O4NPs. The ENPs induced cytotoxicity and nuclear morphological changes in hMSC depending on dose. The cell viability decreases with increase in concentration of ENPs. The cellular morphology studies revealed that ENPs damaged the cells. These preliminary findings have implications for the use of these nanoparticles in food industry with systematic regulations.

Keywords: Cobalt oxide, Human mesenchymal stem cells, MgO, Silver.

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Authors: Elnasharty M., El Sharaby A., Nor El-din A.

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The gustatory system allows animals to distinguish varieties of food and affects greatly the consumption of food, hence the health and growth of animals. In the current study, we investigated the histogenesis of vallate papillae (VLP) in the rabbit tongue using light and scanning electron microscopy. Samples were obtained from rabbit embryos at the embryonic days 16-30 (E16-30), and from newborns until maturity; 6 months. At E16, the first primordia of vallate papillae were observed as small pits on the surface epithelium of the tongue-s root. At E18, the caudal part was prominent with loose mesenchymal tissue core; meanwhile the rostral part of the papilla was remained as a thick mass of epithelial cells. At E20-24, the side epithelium formed the primitive annular groove. At E26, the primitive taste buds appeared only at the papillary surface and reached their maturity by E28. The annular groove started to appear at E26 became more defined at E28. The definitive vallate papillae with substantial number of apparently mature taste buds were observed by the end of the second week. We conclude that the vallate papillae develop early and mature during the early postnatal life.

Keywords: Rabbit, vallate papillae, histogenesis, taste buds.

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

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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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Authors: Liansheng Ren, Xihua Yang, Guoping Wang, Hong Zhang, Lili Zhao, Zhenguo Mi

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The inhibition effect of brazilin to human bladder tumor cell line T24 in vitro and in vivo was studied. The results of the in vitro experiments showed that brazilin has strong inhibition activity on the target cells. The inhibition ratio of 100 μg/mL brazilin and 100 μg/mL mitomycin to the target cells was 90.90 % and 63.24 % respectively, which showed that brazilin has higher inhibition activity than mitomycin under the same concentration. Brazilin could induce cell apoptosis in T24 cells. Significant antitumor activity of brazilin was also showed in the animals experiments. The life extention rate of 200 mg/mL, 300 mg/kg, and 400 mg/kg brazilin intraperitoneally injected into Balb/c-nu-nu nude mice that with human bladder cancer were 51.50 %, 56.90 %, and 58.42 %（P<0.05）. Our study showed that brazilin has significant inhibitory effect on human bladder tumor cell.

Keywords: bladder cancer, brazilin, inhibition, T24 cell line

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

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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, 1α, 25-dihydroxy-vitamin D3

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Authors: Kheireddine El-Boubbou

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Drug delivery to target human acute myeloid leukemia (AML) using a nanoparticulate chemotherapeutic formulation that can deliver drugs selectively to AML cancer is hugely needed. In this work, we report the development of a nanoformulation made of polymeric-stabilized multifunctional magnetic iron oxide nanoparticles (PMNP) loaded with the anticancer drug Doxorubicin (Dox) as a promising drug carrier to treat AML. Dox@PMNP conjugates simultaneously exhibited high drug content, maximized fluorescence, and excellent release properties. Nanoparticulate uptake and cell death following addition of Dox@PMNPs were then evaluated in different types of human AML target cells, as well as on normal human cells. While the unloaded MNPs were not toxic to any of the cells, Dox@PMNPs were found to be highly toxic to the different AML cell lines, albeit at different inhibitory concentrations (IC₅₀ values), but showed very little toxicity towards the normal cells. In comparison, free Dox showed significant potency concurrently to all the cell lines, suggesting huge potentials for the use of Dox@PMNPs as selective AML anticancer cargos. Live confocal imaging, fluorescence and electron microscopy confirmed that Dox is indeed delivered to the nucleus in relatively short periods of time, causing apoptotic cell death. Importantly, this targeted payload may potentially enhance the effectiveness of the drug in AML patients and may further allow physicians to image leukemic cells exposed to Dox@PMNPs using MRI.

Keywords: Magnetic nanoparticles, drug delivery, acute myeloid leukemia, iron oxide, cancer nanotherapy.

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Authors: Parchami A., Fatahian Dehkordi RF.

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The aim of the present investigation was to compare sex differences in thyroid gland structure of rabbits. Five adult male and five adult female (3.1-3.5 kg body weight) New Zealand white rabbits were used in the experiment. Results showed that at light microscopic level, there was no sex difference in microscopic appearance of the thyroid glands. At electron microscopic level, however, the mitochondria and the microvilli of the follicular cells are more numerous and the Golgi complex is also more extensive in male rabbits in comparison to females. Results obtained from micrometric measurements showed that the volume density of the follicles is higher in males than in females, but the differences are not statistically significant .The volume density of epithelium and the height of follicular cells are significantly greater in males than in females and reverse is true about the volume density of interstitium (p<0.05). The volume density of colloid is also greater in females (66±6) than in males (60±7) but the differences are not statistically significant .It was concluded that sex has limited effects on histomorphometric properties of thyroid gland in rabbits.

Keywords: Rabbit, Thyroid Gland, Sex difference, Electron microscope

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

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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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Authors: Andrew Jester, Ross Lee, Pritpal Singh

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As the world transitions to a more sustainable energy economy, the deployment of energy storage technologies is expected to increase to develop a more resilient grid system. However, current technologies are associated with various environmental and safety issues throughout their entire lifecycle; therefore, a new battery technology is desirable for grid applications to curtail these risks. Biological cells, such as human neurons and electrocytes in the electric eel, can serve as a more sustainable design template for a new bio-inspired (i.e., biomimetic) battery. Within biological cells, an electrochemical gradient across the cell membrane forms the membrane potential, which serves as the driving force for ion transport into/out of the cell akin to the charging/discharging of a battery cell. This work serves as the first step for developing such a biomimetic battery cell, starting with the fabrication and characterization of ion-selective membranes to facilitate ion transport through the cell. Performance characteristics (e.g., cell voltage, power density, specific energy, roundtrip efficiency) for the cell under investigation are compared to incumbent battery technologies and biological cells to assess the readiness level for this emerging technology. Using a Na+-Form Nafion-117 membrane, the cell in this work successfully demonstrated behavior like human neurons; these findings will inform how cell components can be re-engineered to enhance device performance.

Keywords: Battery, biomimetic, electrocytes, human neurons, ion-selective membranes, membrane potential.

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Authors: Liangliang Tang, Chang Xu, Xingying Chen

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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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Authors: N. Slepickova Kasalkova, P. Slepicka, L. Bacakova, V. Svorcik

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Tissue engineering includes combination of materials and techniques used for the improvement, repair or replacement of the tissue. Scaffolds, permanent or temporally material, are used as support for the creation of the "new cell structures". For this important component (scaffold), a variety of materials can be used. The advantage of some polymeric materials is their cytocompatibility and possibility of biodegradation. Poly(L-lactic acid) (PLLA) is a biodegradable,  semi-crystalline thermoplastic polymer. PLLA can be fully degraded into H₂O and CO₂. In this experiment, the effect of the surface modification of biodegradable polymer (performed by plasma treatment) on the various cell types was studied. The surface parameters and changes of the physicochemical properties of modified PLLA substrates were studied by different methods. Surface wettability was determined by goniometry, surface morphology and roughness study were performed with atomic force microscopy and chemical composition was determined using photoelectron spectroscopy. The physicochemical properties were studied in relation to cytocompatibility of human osteoblast (MG 63 cells), rat vascular smooth muscle cells (VSMC), and human stem cells (ASC) of the adipose tissue in vitro. A fluorescence microscopy was chosen to study and compare cell-material interaction. Important parameters of the cytocompatibility like adhesion, proliferation, viability, shape, spreading of the cells were evaluated. It was found that the modification leads to the change of the surface wettability depending on the time of modification. Short time of exposition (10-120 s) can reduce the wettability of the aged samples, exposition longer than 150 s causes to increase of contact angle of the aged PLLA. The surface morphology is significantly influenced by duration of modification, too. The plasma treatment involves the formation of the crystallites, whose number increases with increasing time of modification. On the basis of physicochemical properties evaluation, the cells were cultivated on the selected samples. Cell-material interactions are strongly affected by material chemical structure and surface morphology. It was proved that the plasma treatment of PLLA has a positive effect on the adhesion, spreading, homogeneity of distribution and viability of all cultivated cells. This effect was even more apparent for the VSMCs and ASCs which homogeneously covered almost the whole surface of the substrate after 7 days of cultivation. The viability of these cells was high (more than 98% for VSMCs, 89-96% for ASCs). This experiment is one part of the basic research, which aims to easily create scaffolds for tissue engineering with subsequent use of stem cells and their subsequent "reorientation" towards the bone cells or smooth muscle cells.

Keywords: Poly(L-lactic acid), plasma treatment, surface characterization, cytocompatibility, human osteoblasts, rat vascular smooth muscle cells, human stem cells.

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

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

Keywords: Arsenic trioxide, cytotoxixity, genotoxicity, HaCaT.

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Authors: Ibrahim M. S. Shnawa

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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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Authors: Harshani Uggallage, Kapila D. Dissanayaka

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A Sulforhodamine-B assay-guided fractionation on Nigella sativa seeds was conducted to determine the presence of cytotoxic compounds against human hepatoma (HepG2) cells. Initially, a freeze-dried sample of Nigella sativa seeds was sequentially extracted into solvents of increasing polarities. Crude extracts from the sequential extraction of Nigella sativa seeds in chloroform and ethyl acetate showed the highest cytotoxicity. The combined mixture of these two extracts was subjected to bioassay guided fractionation using a modified Kupchan method of partitioning, followed by Sephadex® LH-20 chromatography. This chromatographic separation process resulted in a column fraction with a convincing IC50 (half-maximal inhibitory concentration) value of 13.07 µg/ml, which is considerable for developing therapeutic drug leads against human hepatoma. Reversed phase High-Performance Liquid Chromatography (HPLC) was finally conducted for the same column fraction and the result indicates the presence of one or several main cytotoxic compounds against human HepG2 cells.

Keywords: Cytotoxic compounds, half-maximal inhibitory concentration, high-performance liquid chromatography, human HepG2 cells, Nigella sativa seeds, Sulforhodamine-B assay-guided fractionation.

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Authors: Vahid Anari, Mina Bakhshi

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Visual search and identification of immunohistochemically stained tissue of meningioma was performed manually in pathologic laboratories to detect and diagnose the cancers type of meningioma. This task is very tedious and time-consuming. Moreover, because of cell's complex nature, it still remains a challenging task to segment cells from its background and analyze them automatically. In this paper, we develop and test a computerized scheme that can automatically identify cells in microscopic images of meningioma and classify them into positive (proliferative) and negative (normal) cells. Dataset including 150 images are used to test the scheme. The scheme uses Fuzzy C-means algorithm as a color clustering method based on perceptually uniform hue, saturation, value (HSV) color space. Since the cells are distinguishable by the human eye, the accuracy and stability of the algorithm are quantitatively compared through application to a wide variety of real images.

Keywords: Positive cell, color segmentation, HSV color space, immunohistochemistry, meningioma, thresholding, fuzzy c-means.

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Authors: Elnasharty M. A., Abou-Ghanema I. I., Sayed-Ahmed A., A. Abo Elnour

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The sequential morphologic changes of rabbit duodenal mucosa-submucosa were studied from primodial stage to birth in 15 fetuses and during the early days of life in 21 rabbit newborns till maturity using light, scanning and transmission electron microscopy. Fetal rabbit duodenum develops from a simple tube of stratified epithelium to a tube containing villus and intervillus regions of simple columnar epithelium. By day 21 of gestation, the first rudimentary villi were appeared and by day 24 the first true villi were appeared. The Crypts of Lieberkuhn did not appear until birth. By the first day of postnatal life the duodenal glands appeared. The histological maturity of the rabbit small intestine occurred one month after birth. In conclusion, at all stages, the sequential morphologic changes of the rabbit small intestine developed to meet the structural and physiological demands during the fetal stage to be prepared to extra uterine life.

Keywords: Duodenum, mucosa, submucosa, morphogenesis, rabbit.

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

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

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

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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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Authors: Antonella Bandiera

Abstract:

A new sythetic gene coding for a Human Elastin-Like Polypeptide was constructed and expressed. The recombinant product was tested as coating agent to realize a surface suitable for cell growth. Coatings showed peculiar features and different human cell lines were seeded and cultured. All cell lines tested showed to adhere and proliferate on this substrate that has been shown also to exert a specific effect on cells, depending on cell type.

Keywords: elastin, recombinant protein, coating, cell adhesion.

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Authors: Amaneh Javid, Shahin Ahmadian, Ali A. Saboury, Saeed Rezaei-Zarchi

Abstract:

This study determines the effect of naked and heparinbased super-paramagnetic iron oxide nanoparticles on the human cancer cell lines of A2780. Doxorubicin was used as the anticancer drug, entrapped in the SPIO-NPs. This study aimed to decorate nanoparticles with heparin, a molecular ligand for 'active' targeting of cancerous cells and the application of modified-nanoparticles in cancer treatment. The nanoparticles containing the anticancer drug DOX were prepared by a solvent evaporation and emulsification cross-linking method. The physicochemical properties of the nanoparticles were characterized by various techniques, and uniform nanoparticles with an average particle size of 110±15 nm with high encapsulation efficiencies (EE) were obtained. Additionally, a sustained release of DOX from the SPIO-NPs was successful. Cytotoxicity tests showed that the SPIO-DOX-HP had higher cell toxicity than the individual HP and confocal microscopy analysis confirmed excellent cellular uptake efficiency. These results indicate that HP based SPIO-NPs have potential uses as anticancer drug carriers and also have an enhanced anticancer effect.

Keywords: Heparin, A2780 cells, ovarian cancer, nanoparticles, doxorubicin.

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Authors: Faten. A. Khorshid, Gehan. A. Raouf, Salem. M. El-Hamidy, Gehan. S. Al-amri, Nourah. A. Alotaibi, Taha A. Kumosani

Abstract:

Cancer becomes one of the leading cause of death in many countries over the world. Fourier-transform infrared (FTIR) spectra of human lung cancer cells (A549) treated with PMF (natural product extracted from PM 701) for different time intervals were examined. Second derivative and difference method were taken in comparison studies. Cesium (Cs) and Rubidium (Rb) nanoparticles in PMF were detected by Energy Dispersive X-ray attached to Scanning Electron Microscope SEM-EDX. Characteristic changes in protein secondary structure, lipid profile and changes in the intensities of DNA bands were identified in treated A549 cells spectra. A characteristic internucleosomal ladder of DNA fragmentation was also observed after 30 min of treatment. Moreover, the pH values were significantly increases upon treatment due to the presence of Cs and Rb nanoparticles in the PMF fraction. These results support the previous findings that PMF is selective anticancer agent and can produce apoptosis to A549 cells.

Keywords: Apoptosis, FTIR spectroscopy, pH therapy, Scanning Electron Microscope- Energy Dispersive X-ray (SEMEDX).

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Authors: M. Baryshev, D.Vainauska, S. Kozireva, A.Karpovs

Abstract:

Liposomal magnetofection is the most powerful nonviral method for the nucleic acid delivery into the cultured cancer cells and widely used for in vitro applications. Use of the static magnetic field condition may result in non-uniform distribution of aggregate complexes on the surface of cultured cells. To prevent this, we developed the new device which allows to concentrate aggregate complexes under dynamic magnetic field, assisting more contact of these complexes with cellular membrane and, possibly, stimulating endocytosis. Newly developed device for magnetofection under dynamic gradient magnetic field, “DynaFECTOR", was used to compare transfection efficiency of human liver hepatocellular carcinoma cell line HepG2 with that obtained by lipofection and magnetofection. The effect of two parameters on transfection efficiency, incubation time under dynamic magnetic field and rotation frequency of magnet, was estimated. Liposomal magnetofection under dynamic gradient magnetic field showed the highest transfection efficiency for HepG2 cells.

Keywords: Dynamic magnetic field, Lipofection, Magnetofection

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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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Authors: Archana Mukherjee, Aruna Korde, Sudipta Chakraborty, H. D. Sarma, Grace Samuel, M. R. A. Pillai

Abstract:

Radiolabeled cyclic RGD peptides targeting integrin αvβ3 are reported as promising agents for the early diagnosis of metastatic tumors. With an aim to improve tumor uptake and retention of the peptide, cyclic RGD peptide dimer E[c (RGDfK)] 2 (E = Glutamic acid, f = phenyl alanine, K = lysine) coupled to the bifunctional chelator DOTA was custom synthesized and radiolabelled with 68Ga. Radiolabelling of cyclic RGD peptide dimer with 68Ga was carried out using HEPES buffer and biological evaluation of the complex was done in nude mice bearing HT29 tumors.

Keywords: 68Ga peptides, Angiogenesis imaging, Cyclic RGD peptides, PET Imaging.

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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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Authors: P. Narrima, C. Y. Looi, M. A. Mohd, H. M. Ali

Abstract:

Persea declinata (Bl.) Kosterm is a member of the Lauraceae family, widely distributed in Southeast Asia. It is from the same genus with avocado (Persea americana Mill), which is widely consumed as food and for medicinal purposes. In the present study, we examined the anticancer properties of Persea declinata (Bl.) Kosterm bark methanolic crude extract (PDM). PDM exhibited a potent antiproliferative effect in MCF-7 human breast cancer cells, with an IC50 value of 16.68 .g/mL after 48h of treatment. We observed that PDM caused cell cycle arrest and subsequent apoptosis in MCF-7 cells, as exhibited by increased population at G0/G1 phase, higher lactate dehydrogenase (LDH) release, and DNA fragmentation. Mechanistic studies showed that PDM caused significant elevation in ROS production, leading to perturbation of mitochondrial membrane potential, cell permeability, and activation of caspases-3/7. On the other hand, real-time PCR and Western blot analysis showed that PDM treatment increased the expression of the proapoptotic molecule, Bax, but decreased the expression of prosurvival proteins, Bcl-2 and Bcl-xL, in a dose-dependent manner. These findings imply that PDM could inhibit proliferation in MCF-7 cells via cell cycle arrest and apoptosis induction, indicating its potential as a therapeutic agent worthy of further development.

Keywords: Antiproliferative, apoptosis, MCF-7 human breast cancer, Persea declinata.

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