Search results for: mouse embryonic fibroblast.

Authors: Priyanka Gupta, Paul Verma, Kerry Hourigan, Jayesh Bellare, Sameer Jadhav

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Understanding the consumption and production of various metabolites of fibroblast conditioned media is needed for its proper and optimized use in expansion of pluripotent stem cells. For this purpose, we have used the HPLC method to analyse the consumption of glucose and the production of lactate over time by mouse embryonic fibroblasts. The experimental data have also been compared with mathematical model fits. 0.025 moles of lactate was produced after 72 hrs while the glucose concentration decreased from 0.017 moles to 0.011 moles. The mathematical model was able to predict the trends of glucose consumption and lactate production.

Keywords: Conditioned media, HPLC, metabolite analysis, mouse embryonic fibroblast.

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Authors: Kasem Theerakittayakorn, Tanom Bunprasert

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Mouse L929 fibroblastic cell line, which is widely used in many experiment aspects, was tested for their differentiation potency in osteogenic differentiation and adipogenic differentiation. Human dermal fibroblasts, which their differentiation potency are still be in confliction, also be taken in the experiment. The differentiations were conducted by using the inducing medium ingredients which is generally used to induce differentiation of stem cells. By the inducing media used, L929 mouse fibroblasts successfully underwent osteogenic differentiation and adipogenic differentiation while human dermal fibroblasts underwent only osteogenic differentiation but not for adipogenic differentiation. Human dermal fibroblasts are hard to be differentiated in adipogenic lineage and need specific proper condition for induction.

Keywords: Adipogenic differentiation, Fibroblast, Inducingmedium, Osteogenic differentiation

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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: H. Mhamdi, N. Hamrouni, A. Temimi, M. Bouhlel

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In this article, we expose our research work in Human-machine Interaction. The research consists in manipulating the workspace by eyes. We present some of our results, in particular the detection of eyes and the mouse actions recognition. Indeed, the handicaped user becomes able to interact with the machine in a more intuitive way in diverse applications and contexts. To test our application we have chooses to work in real time on videos captured by a camera placed in front of the user.

Keywords: Computer vision, Face and Eyes Detection, Mouse pointer recognition.

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Authors: Mouse, cardiac myocytes, computer simulation, action potential.

Abstract:

Heterogeneous repolarization causes dispersion of the T-wave and has been linked to arrhythmogenesis. Such heterogeneities appear due to differential expression of ionic currents in different regions of the heart, both in healthy and diseased animals and humans. Mice are important animals for the study of heart diseases because of the ability to create transgenic animals. We used our previously reported model of mouse ventricular myocytes to develop 2D mouse ventricular tissue model consisting of 14,000 cells (apical or septal ventricular myocytes) and to study the stability of action potential propagation and Ca2+ dynamics. The 2D tissue model was implemented as a FORTRAN program code for highperformance multiprocessor computers that runs on 36 processors. Our tissue model is able to simulate heterogeneities not only in action potential repolarization, but also heterogeneities in intracellular Ca2+ transients. The multicellular model reproduced experimentally observed velocities of action potential propagation and demonstrated the importance of incorporation of realistic Ca2+ dynamics for action potential propagation. The simulations show that relatively sharp gradients of repolarization are predicted to exist in 2D mouse tissue models, and they are primarily determined by the cellular properties of ventricular myocytes. Abrupt local gradients of channel expression can cause alternans at longer pacing basic cycle lengths than gradual changes, and development of alternans depends on the site of stimulation.

Keywords: Mouse, cardiac myocytes, computer simulation, action potential

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Authors: Arslan Qamar Malik, Jehanzeb Ahmad

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The paper presents a novel idea to control computer mouse cursor movement with human eyes. In this paper, a working of the product has been described as to how it helps the special people share their knowledge with the world. Number of traditional techniques such as Head and Eye Movement Tracking Systems etc. exist for cursor control by making use of image processing in which light is the primary source. Electro-oculography (EOG) is a new technology to sense eye signals with which the mouse cursor can be controlled. The signals captured using sensors, are first amplified, then noise is removed and then digitized, before being transferred to PC for software interfacing.

Keywords: Human Computer Interaction, Real-Time System, Electro-oculography, Signal Processing.

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Authors: Changsoo Chon, Sangkuy Han, Donghyun Seo, Jihyung Park, Bokku Kang, Hansung Kim, Keyoungjin Chun, Cheolwoong Ko

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The purpose of the study is to develop a finite element model based on 3D bone structural images of Micro-CT and to analyze the stress distribution for the osteoporosis mouse femora. In this study, results of finite element analysis show that the early osteoporosis of mouse model decreased a bone density in trabecular region; however, the bone density in cortical region increased.

Keywords: Micro-CT, finite element analysis, osteoporosis, bone strength.

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Authors: A. Doosti, S. Moshkelani

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Brucellosis is a zoonotic disease; its symptoms and appearances are not exclusive in human and its traditional diagnosis is based on culture, serological methods and conventional PCR. For more sensitive, specific detection and differentiation of Brucella spp., the real time PCR method is recommended. This research has performed to determine the presence and prevalence of Brucella spp. and differentiation of Brucella abortus and Brucella melitensis in house mouse (Mus musculus) in west of Iran. A TaqMan analysis and single-step PCR was carried out in total 326 DNA of Mouse's spleen samples. From the total number of 326 samples, 128 (39.27%) gave positive results for Brucella spp. by conventional PCR, also 65 and 32 out of the 128 specimens were positive for B. melitensis, B. abortus, respectively. These results indicate a high presence of this pathogen in this area and that real time PCR is considerably faster than current standard methods for identification and differentiation of Brucella species. To our knowledge, this study is the first prevalence report of direct identification and differentiation of B. abortus and B. melitensis by real time PCR in mouse tissue samples in Iran.

Keywords: Differentiation, B. abortus, B. melitensis, TaqManprobe, Iran.

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Authors: Hyun June Kim, Tailong Shi, Seden Akdagli, Sam Most, Yuling Yan

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Quantitative analyses of whisker movements provide a means to study functional recovery and regeneration of mouse facial nerve after an injury. However, accurate tracking of the mouse whisker movement is challenging. Most methods for whisker tracking require manual intervention, e.g. fixing the head of the mouse during a study. Here we describe a semi-automated image processing method, which is applied to high-speed video recordings of free-moving mice to track the whisker movements. We first track the head movement of a mouse by delineating the lower head contour frame-by-frame that allows for detection of the location and orientation of the head. Then, a region of interest is identified for each frame; the subsequent application of a mask and the Hough transform detects the selected whiskers on each side of the head. Our approach is used to examine the functional recovery of damaged facial nerves in mice over a course of 21 days.

Keywords: Mystacial macrovibrissae, whisker tracking, head tracking, facial nerve recovery.

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Authors: V. Tawiwat, S. Sarawut

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Applied a mouse-s roller with a gripper to increase the efficiency for a gripper can learn to a material handling without slipping. To apply a gripper, we use the optimize principle to develop material handling by use a signal for checking a roller mouse that rotate or not. In case of the roller rotates means that the material slips. A gripper will slide to material handling until the roller will not rotate. As this experiment has test material handling for comparing a grip force that uses to material handling of the 10-human with the applied gripper. We can summarize that human exert the material handling more than the applied gripper. Because of the gripper can exert more befit to material handling than human and may be a minimum force to lift a material without slipping.

Keywords: Optimize, Gripper, Mouse's Roller, Minimum Force.

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Authors: Ching-Sheng Wang, Lun-Ping Hung, Sheng-Yu Peng, Li-Chieh Cheng

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The computer has become an essential tool in modern life, and the combined use of a computer with a projector is very common in teaching and presentations. However, as typical computer operating devices involve a mouse or keyboard, when making presentations, users often need to stay near the computer to execute functions such as changing pages, writing, and drawing, thus, making the operation time-consuming, and reducing interactions with the audience. This paper proposes a laser pointer interaction system able to simulate mouse functions in order that users need not remain near the computer, but can directly use laser pointer operations from at a distance. It can effectively reduce the users- time spent by the computer, allowing for greater interactions with the audience.

Keywords: laser pointer, presentation, interaction, mousefunction

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Authors: M. H. Rajikin, S. M. M. Syairah, A. R. Sharaniza

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Effects of nicotine on pre-partum body weight and preimplantation embryonic development has been reported previously. Present study was conducted to determine the effects of annatto (Bixa orellana)-derived delta-tocotrienol (TCT) (with presence of 10% gamma-TCT isomer) on the nicotine-induced reduction in body weight and 8-cell embryonic growth in mice. Twenty-four 6-8 weeks old (23-25g) female balb/c mice were randomly divided into four groups (G1-G4; n=6). Those groups were subjected to the following treatments for 7 consecutive days: G1 (control) were gavaged with 0.1 ml tocopherol stripped corn oil. G2 was subcutaneously (s.c.) injected with 3 mg/kg/day of nicotine. G3 received concurrent treatment of nicotine (3 mg/kg/day) and 60 mg/kg/day of δ-TCT mixture (contains 90% delta & 10% gamma isomers) and G4 was given 60 mg/kg/day of δ-TCT mixture alone. Body weights were recorded daily during the treatment. On Day 8, females were superovulated with 5 IU Pregnant Mare’s Serum Gonadotropin (PMSG) for 48 hours followed with 5 IU human Chorionic Gonadotropin (hCG) before mated with males at the ratio of 1:1. Females were sacrificed by cervical dislocation for embryo collection 48 hours post-coitum. Collected embryos were cultured in vitro. Results showed that throughout Day 1 to Day 7, the body weight of nicotine treated group (G2) was significantly lower (p<0.05) than that of G1, G3 and G4. Intervention with δ-TCT mixture (G3) managed to increase the body weight close to the control group. This is also observed in the group treated with δ-TCT mixture alone (G4). The development of 8-cell embryos following in vitro culture (IVC) was totally inhibited in G2. Intervention with δ- TCT mixture (G3) resulted in the production of 8-cell embryos, although it was not up to that of the control group. Treatment with δ- TCT mixture alone (G4) caused significant increase in the average number of produced 8-cell embryo compared to G1. Present data indicated that δ-TCT mixture was able to reverse the body weight loss in nicotine treated mice and the development of 8-cell embryos was also improved. Further analysis on the quality of embryos need to done to confirm the effects of δ-TCT mixture on preimplantation embryos.

Keywords: δ-tocotrienol, body weight, nicotine, preimplantation embryonic development.

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Authors: Kasun Karunanayaka, Sanath Siriwardana, Chamari Edirisinghe, Ryohei Nakatsu, PonnampalamGopalakrishnakone

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In this paper, we are presenting a new type of pointing interface for computers which provides mouse functionalities with near surface haptic feedback. Further, it can be configured as a haptic display where users may feel the basic geometrical shapes in the GUI by moving the finger on top of the device surface. These functionalities are achieved by tracking three dimensional positions of the neodymium magnet using Hall Effect sensors grid and generating like polarity haptic feedback using an electromagnet array. This interface brings the haptic sensations to the 3D space where previously it is felt only on top of the buttons of the haptic mouse implementations.

Keywords: Pointing interface, near surface haptic feedback, tactile display, tangible user interface.

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Authors: Sri Sumartiningsih

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In elevating performance in competetive sports, an athlete must continously train in achieving maximum performance,but needs to pay attention to recovery therapy, that is to recover from fatigue as well as injury.The correct recovery therapy will assist in process of recovery and helps in the training in achieving better performace. Binahong (Anredera cordifolia) was proven empirically by the locals in assisting speedy recovery from an injury.Clinical research with lab animals receiving blunt trauma injury, microscopically shown signs of: 1) redness, 2) heatiness, 3) swelling and, 4) lack of activity. There is also microscopic indication of: 1) infiltration of inflame cells (migration of cells to the trauma area), 2) Cells necrosis, 3) Congestion (as a result of dead red blood cells), 4) uedema. On administration of Binahong for 3 days, there is a significant drop of 5% in cell inflammation, 2% increase of fibroblast (cell membrance) count.Conclutin: Binahong do assist in reducing cell inflammation and increase counts of cells fibroblast. Suggestion: In helping athlete's to recover from force injury, we need study about Binahong's roots to inflammation cell and healing of injuried cell.

Keywords: Binahong, sport injury, hematoma

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Authors: Sansani Rungrattawatchai

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The White Kwao (Pueraria mirifica), a potent phytoestrogenic medicinal plant, has long been use in Thailand as a traditional folkmedicine. However, no scientific information of the direct effect of White Kwao on the development of mammalian embryo was available. Therefore, the purpose of this study was to investigate the effect of White Kwao extract on the in vitro development and implantation rate of mouse embryos. This study was designed into two experiments. In the first experiment, the two-cell stage mouse embryos were collected from the oviduct of superovulated mature female mice, and randomly cultured in three different media, the M16, M16 supplemented with 0.52μg esthinylestradiol-17β, and M16 supplemented with 10 mg/ml White Kwao extract. The culture was incubated in CO2 incubator at 37 oC . After the embryos were cultivated, the developments of embryos were observed every 24 hours for 5 days. The development rate of embryos from the two-cell stage to blastocyst stage in the media was with White Kwao was significantly higher (p<0.05) than those of the control group (68.50% versus 43.50%) but did not differ from the positive control group (68.50% versus 57.66%). In the second experiment, hatched blastocysts, which obtained from three different media, were differently labeled the nuclei with two polynucleotide-specific fluorochromes, the propidium iodide (PI) and the bisbenzimide. The results showed that the number of trophectoderm cells in the blastocysts that cultivated in the media with White Kwao did not significantly differ from the control (80.00 versus 70 cells) and the positive control group (80.00 versus 112.50 cells). The average number of inner cell mass in the White Kwao treated group did not significantly differ from the control group (20.50 versus 16.00 cells) and the positive control group (20.50 versus 20.50 cells). The total cell number including the trophectoderm and the inner cell mass of the individual hatched blastocyst was evaluated. The cell number in the blastocysts obtained from the media with the White Kwao did not significantly differ from the control (94.25 + 9.50 versus 92.33 + 4.05) and the positive control group (94.25 + 9.50 versus 110.33 + 9.16). The results demonstrated that the White Kwao treatment group did have a stimulating effect on the in vitro development of mouse embryos. The exact mechanism that White Kwao stimulated mouse embryo development is not known. The suspect mechanism may in a manner similar to the mechanism that of estrogen stimulated the development of the mouse embryos. Futher studies are needed to transfer the blastocyst into the endometrium of pseudopreagnancy mice to evaluate the effect of White Kwao on implantation

Keywords: White Kwao (Pueraria mirifica), blastocyst.

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Authors: Nadezhda M. Usmanova, Nikolai V. Tomilin

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Mammalian genomes contain large number of retroelements (SINEs, LINEs and LTRs) which could affect expression of protein coding genes through associated transcription factor binding sites (TFBS). Activity of the retroelement-associated TFBS in many genes is confirmed experimentally but their global functional impact remains unclear. Human SINEs (Alu repeats) and mouse SINEs (B1 and B2 repeats) are known to be clustered in GCrich gene rich genome segments consistent with the view that they can contribute to regulation of gene expression. We have shown earlier that Alu are involved in formation of cis-regulatory modules (clusters of TFBS) in human promoters, and other authors reported that Alu located near promoter CpG islands have an increased frequency of CpG dinucleotides suggesting that these Alu are undermethylated. Human Alu and mouse B1/B2 elements have an internal bipartite promoter for RNA polymerase III containing conserved sequence motif called B-box which can bind basal transcription complex TFIIIC. It has been recently shown that TFIIIC binding to B-box leads to formation of a boundary which limits spread of repressive chromatin modifications in S. pombe. SINEassociated B-boxes may have similar function but conservation of TFIIIC binding sites in SINEs located near mammalian promoters has not been studied earlier. Here we analysed abundance and distribution of retroelements (SINEs, LINEs and LTRs) in annotated sequences of the Database of mammalian transcription start sites (DBTSS). Fractions of SINEs in human and mouse promoters are slightly lower than in all genome but >40% of human and mouse promoters contain Alu or B1/B2 elements within -1000 to +200 bp interval relative to transcription start site (TSS). Most of these SINEs is associated with distal segments of promoters (-1000 to -200 bp relative to TSS) indicating that their insertion at distances >200 bp upstream of TSS is tolerated during evolution. Distribution of SINEs in promoters correlates negatively with the distribution of CpG sequences. Using analysis of abundance of 12-mer motifs from the B1 and Alu consensus sequences in genome and DBTSS it has been confirmed that some subsegments of Alu and B1 elements are poorly conserved which depends in part on the presence of CpG dinucleotides. One of these CpG-containing subsegments in B1 elements overlaps with SINE-associated B-box and it shows better conservation in DBTSS compared to genomic sequences. It has been also studied conservation in DBTSS and genome of the B-box containing segments of old (AluJ, AluS) and young (AluY) Alu repeats and found that CpG sequence of the B-box of old Alu is better conserved in DBTSS than in genome. This indicates that Bbox- associated CpGs in promoters are better protected from methylation and mutation than B-box-associated CpGs in genomic SINEs. These results are consistent with the view that potential TFIIIC binding motifs in SINEs associated with human and mouse promoters may be functionally important. These motifs may protect promoters from repressive histone modifications which spread from adjacent sequences. This can potentially explain well known clustering of SINEs in GC-rich gene rich genome compartments and existence of unmethylated CpG islands.

Keywords: Retroelement, promoter, CpG island, DNAmethylation.

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Authors: Junki Miyamoto, Mariko Kiyomi, Yuuki Nagashio, Takuya Suzuki, Soichi Tanabe

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Anti-allergic effects of royal jelly were evaluated in a human-like mouse model of atopic dermatitis. NC/Nga mice were cutaneously applied with royal jelly for 6 weeks. Royal jelly-treated mice exhibited lower levels of serum total immunoglobulin E in comparison with controls. We found that the treatment decreased (11% to the control) expression of mRNA for aquaporin-3, which is involved in the modulation of epidermal hydration. Microarray analysis revealed more than 10-fold changes in the expression of several genes, such as transglutaminase 2, repetin, and keratins. In normal human epidermal keratinocytes, royal jelly extract suppressed interleukin-8 elevation induced by TNF-α and interferon-γ, suggesting direct anti-inflammatory activity in keratinocytes. Collectively, topical application of royal jelly may be useful for amelioration of lesions and inflammation in atopic dermatitis.

Keywords: Aquaporin 3, immunoglobulin E, NC/Nga, royal jelly.

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Authors: M. Mousavi-Daramoroudi, H. Yousefnia, F. Abbasi-Davani, S. Zolghadri, S. Kakaei

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Despite the appropriate characteristics of ¹⁷⁷Lu and DOTATOC, to our best knowledge, the therapeutic benefit of ¹⁷⁷Lu-DOTATOC complex in breast cancer has not been reported until now. In this study, biodistribution of ¹⁷⁷Lu-DOTA-TOC in mouse tumor model for evaluation of possible utilization of this complex in breast cancer treatment was investigated.¹⁷⁷Lu was prepared with the specific activity of 2.6-3 GBq.mg^-1 and radionuclidic purity higher than 99%. The radiolabeled complex was prepared in the optimized conditions with the radiochemical purity higher than 99%. The final solution was injected to the BALB/c mice with adenocarcinoma breast cancer. The biodistribution results showed major accumulation in the kidneys as the major excretion route and the somatostatin receptor-positive tissues such as pancreas compared with the other tissues. Also, significant uptake was observed in tumor even in longer time after injection. According to the results obtained in this research study, somatostatin receptors expressed in breast cancers can be targeted with DOTATOC analogues especially with ¹⁷⁷Lu-DOTATOC as an ideal therapeutic agent.

Keywords: 177Lu, DOTATOC, adenocarcinoma, breast cancer, BALB/c mice.

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Authors: M. Jarrar, S. Behl, N. Shaheen, A. Fatima, R. Nasab

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Skin aging is a slow multifactorial process influenced by both internal as well as external factors. Ultra-violet radiations (UV), diet, smoking and personal habits are the most common environmental factors that affect skin aging. Fat contents and fibrous proteins as collagen and elastin are core internal structural components. The direct influence of UV on elastin integrity and health is central on aging of skin especially by time. The deposition of abnormal elastic material is a major marker in a photo-aged skin. Searching for compounds that may protect against cutaneous photodamage is exceedingly valued. Retinoids and alpha hydroxy acids have been endorsed by some researchers as possible candidates for protecting and or repairing the effect of UV damaged skin. For consolidating a better system of anti- and protective effects of such anti-aging agents, we evaluated the combinatory effects of various dosages of lactic acid and retinol on the dermal fibroblast’s elastin levels exposed to UV. The UV exposed cells showed significant reduction in the elastin levels. A combination of drugs with a higher concentration of lactic acid (30 -35 mM) and a lower concentration of retinol (10-15mg/mL) showed to work better in maintaining elastin concentration in UV exposed cells. We assume this preservation could be the result of increased tropo-elastin gene expression stimulated by retinol whereas lactic acid probably repaired the UV irradiated damage by enhancing the amount and integrity of the elastin fibers.

Keywords: Alpha Hydroxy Acid, Elastin, Retinol, Ultraviolet radiations.

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Authors: S. M. M. Syairah, M. H. Rajikin, A-R. Sharaniza

Abstract:

Several embryonic cellular mechanism including cell cycle, growth and apoptosis are regulated by phosphatidylinositol-3- kinase (PI3K)/Akt signaling pathway. The goal of present study is to determine the effects of annatto (Bixa orellana)-derived δ-tocotrienol (δ-TCT) on the regulations of PI3K/Akt genes in murine morula. Twenty four 6-8 week old (23-25g) female balb/c mice were randomly divided into four groups (G1-G4; n=6). Those groups were subjected to the following treatments for 7 consecutive days: G1 (control) received tocopherol stripped corn oil, G2 was given 60 mg/kg/day of δ-TCT mixture (contains 90% delta & 10% gamma isomers), G3 was given 60 mg/kg/day of pure δ-TCT (>98% purity) and G4 received 60 mg/kg/day α-TOC. On Day 8, females were superovulated with 5 IU Pregnant Mare’s Serum Gonadotropin (PMSG) for 48 hours followed with 5 IU human Chorionic Gonadotropin (hCG) before mated with males at the ratio of 1:1. Females were sacrificed by cervical dislocation for embryo collection 48 hours post-coitum. About fifty morulas from each group were used in the gene expression analyses using Affymetrix QuantiGene Plex 2.0 Assay. Present data showed a significant increase (p<0.05) in the average number (mean + SEM) of morula produced in G2 (27.32 + 0.23), G3 (25.42 + 0.21) and G4 (27.21 + 0.34) compared to control group (G1 – 14.61 + 0.25). This is parallel with the high expression of PDK1 gene with increase of 2.75-fold (G2), 3.07-fold (G3) and 3.59-fold (G4) compared to G1. From the present data, it can be concluded that supplementation with δ-TCT(s) and α-TOC induced high expression of PDK1 in G2-G4 which enhanced the PI3K/Akt signaling activity, resulting in the increased number of morula.

Keywords: Embryonic development, morula, nicotine, vitamin E.

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Authors: Eszter Tóth, Ervin Welker

Abstract:

Shadoo protein (Sho) was described in 2003 as the newest member of Prion protein superfamily [1]. Sho has similar structural motifs like prion protein (PrP) that is known for its central role in transmissible spongiform enchephalopathies. Although a great number of functions have been proposed, the exact physiological function of PrP is not known yet. Investigation of the function and localization of Sho may help us to understand the function of the Prion protein superfamily. Analyzing the subcellular localization of YFP-tagged forms of Sho, we detected the protein in the plasma membrane and in the nucleus of various cell lines. To reveal the localization of the endogenous protein we generated antibodies against Shadoo as well as employed commercially available anti-Shadoo antibodies: i) EG62 anti-mouse Shadoo antibody generated by Eurogentec Ltd.; ii) S-12 anti-human Shadoo antibody by Santa Cruz Biotechnology Inc.; iii) R-12 anti-mouse Shadoo antibody by Santa Cruz Biotechnology Inc.; iv) SPRN antibody against human Shadoo by Abgent Inc. We carried out immunocytochemistry on non-transfected HeLa, Zpl 2-1, Zw 3-5, GT1-1, GT1-7 and SHSY5Y cells as well as on YFP-Sho, Sho-YFP, and YFP-GPI transfected HeLa cells. Their specificity (in antibody-peptide competition assay) and co-localization (with the YFP signal) were assessed.

Keywords: Shadoo, prion protein, immunocytochemistry, antibody-peptide competition assay, antibody.

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Authors: Shanshan Li

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Beginning with the debate of concept, this essay explains the historical source and development of flexible housing in China. In the former part, the flexibility contained in traditional house is explored. While in the latter, the relevant practices in modern times are systematically analyzed as three phases–the Embryonic Period (1949 - 1980), the Systematic Practice (1981 - 2000), as well as the Integrated Trend and Prosperity (2001 - present). As a conclusion, the generalized flexibility is tentatively discussed.

Keywords: Flexibility, Long-term effectiveness, Variety, Social background.

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Authors: Samaneh Nazeri, Bagher Mojazi Amiri, Hamid Farahmand

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Production of high quality fish eggs with reasonable hatching rate makes a success in aquaculture industries. It is influenced by the environmental stimulators and inhibitors. Diazinon is a widely-used pesticide in Golestan province (Southern Caspian Sea, North of Iran) which is washed to the aquatic environment (3 mg/L in the river). It is little known about the effect of this pesticide on the embryogenesis of sturgeon fish, the valuable species of the Caspian Sea. Hormonal content of the egg is an important factor to guaranty the successful passes of embryonic stages. In this study, the fate of Persian sturgeon embryo to 24, 48, 72, and 96-hours exposure of diazinon (LC₅₀ dose) was tested. Also, the effect of thyroid hormones (T3 and T4) on these embryos was tested concurrently or separately with diazinon LC ₅₀ dose. Fertilized eggs are exposed to T3 (low dose: 1 ng/ml, high dose: 10 ng/ml), T4 (low dose: 1 ng/ml, high dose: 10 ng/ml). Six eggs were randomly selected from each treatment (with three replicates) in five developmental stages (two cell- division, neural, heart present, heart beaten, and hatched larvae). The possibility of changing T3, T4, and cortisol contents of the embryos were determined in all treated groups and in every mentioned embryonic stage. The hatching rate in treated groups was assayed at the end of the embryogenesis to clarify the effect of thyroid hormones and diazinon. The results indicated significant differences in thyroid hormone contents, but no significant differences were recognized in cortisol levels at various early life stages of embryos. There was also significant difference in thyroid hormones in (T3, T4) + diazinon treated embryos (P˂0.05), while no significant difference between control and treatments in cortisol levels was observed. The highest hatching rate was recorded in HT3 treatment, while the lowest hatching rate was recorded for diazinon LC₅₀ treatment. The result confirmed that Persian sturgeon embryo is less sensitive to diazinon compared to teleost embryos, and thyroid hormones may increase hatching rate even in the presence of diazinon.

Keywords: Persian sturgeon, diazinon, thyroid hormones, cortisol, embryo.

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Authors: Mustafa M. Donma, Orkide Donma

Abstract:

Obesity in children particularly draws attention, because it may threaten the individual’s future life due to many chronic diseases it may lead to. Most of these diseases including obesity itself altogether are related to inflammation. For this reason, inflammation-related parameters gain importance. Within this context, complete blood cell counts, ratios or indices derived from these counts have recently found some platform to be used as inflammatory markers. So far, mostly adipokines were investigated within the field of obesity. Metabolic inflammation is closely associated with cellular dysfunction. In this study, hematologic inflammatory markers and cytokines produced predominantly by the liver (fibroblast growth factor-21 (FGF-21) and fetuin A) were investigated in pediatric obesity. Two groups were constituted from 76 obese children based on World Health Organization criteria. Group 1 was composed of children, whose age- and sex-adjusted body mass index (BMI) percentiles were between 95 and 99. Group 2 consists of children, who are above 99th percentile. The first and the latter groups were defined as obese (OB) and morbid obese (MO). Anthropometric measurements of the children were performed. Informed consent forms and the approval of the institutional ethics committee were obtained. Blood cell counts and ratios were determined by automated hematology analyzer. The related ratios and indexes were calculated. Statistical evaluation of the data was performed by SPSS program. There was no statistically significant difference in terms of neutrophil-to lymphocyte ratio, monocyte-to-high density lipoprotein cholesterol ratio and platelet-to-lymphocyte ratio between the groups. Mean platelet volume and platelet distribution width values were decreased (p < 0.05), total platelet count, red cell distribution width (RDW) and systemic immune inflammation index values were increased (p < 0.01) in MO group. Both hepatokines were increased in the same group, however increases were not statistically significant. In this group, also a strong correlation was calculated between FGF-21 and RDW when controlled by age, hematocrit, iron and ferritin (r = 0.425; p < 0.01). In conclusion, the association between RDW, a hematologic inflammatory marker, and FGF-21, an inflammation-related hepatokine, found in MO group is an important finding discriminating between OB and MO children. This association is even more powerful when controlled by age and iron-related parameters.

Keywords: Childhood obesity, fetuin A, fibroblast growth factor-21, hematologic markers, red cell distribution width.

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Authors: Kakali Bhadra

Abstract:

Harmalol administration caused remarkable reduction in proliferation of HepG₂ cells with GI₅₀ of 14.2 mM, without showing much cytotoxicity in embryonic liver cell line, WRL-68. Data from circular dichroism and differential scanning calorimetric analysis of harmalol-CT DNA complex shows conformational changes with prominent CD perturbation and stabilization of CT DNA by 8 ^oC. Binding constant and stoichiometry was also calculated using the above biophysical techniques. Further, dose dependent apoptotic induction ability of harmalol was studied in HepG₂ cells using different biochemical assays. Generation of ROS, DNA damage, changes in cellular external and ultramorphology, alteration of membrane, formation of comet tail, decreased mitochondrial membrane potential and a significant increase in Sub G_o/G₁ population made the cancer cell, HepG₂, prone to apoptosis. Up regulation of p53 and caspase 3 further indicated the apoptotic role of harmalol.

Keywords: Apoptosis, beta carboline alkaloid, comet assay, cytotoxicity, ROS.

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Authors: Ling-fei Xu, Qi Zhao, Yan-ru Chen, Mu-chun Zhou, Meng Zhang, Shi-xue Xu

Abstract:

SVM ( Support Vector Machine ) is a new method in the artificial neural network ( ANN ). In the steel making, how to use computer to predict the end point of BOF accuracy is a great problem. A lot of method and theory have been claimed, but most of the results is not satisfied. Now the hot topic in the BOF end point predicting is to use optical way the predict the end point in the BOF. And we found that there exist some regular in the characteristic curve of the flame from the mouse of pudding. And we can use SVM to predict end point of the BOF, just single spectrum intensity should be required as the input parameter. Moreover, its compatibility for the input space is better than the BP network.

Keywords: SVM, predict, BOF, single spectrum intensity.

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Authors: Santosh Kumar, Anand Prakash, Kanak Sinha, Anita K Verma

Abstract:

Vernonia divergens Benth., commonly known as “Insulin Plant” (Fam: Asteraceae) is a potent sugar killer. Locally the leaves of the plant, boiled in water are successfully administered to a large number of diabetic patients. The present study evaluates the putative anti-diabetic ingredients, isolated from the in vivo and in vitro grown plantlets of V. divergens for their antimicrobial and anticancer activities. Sterilized explants of nodal segments were cultured on MS (Musashige and Skoog, 1962) medium in presence of different combinations of hormones. Multiple shoots along with bunch of roots were regenerated at 1mg l-1 BAP and 0.5 mg l-1 NAA. Micro-plantlets were separated and sub-cultured on the double strength (2X) of the above combination of hormones leading to increased length of roots and shoots. These plantlets were successfully transferred to soil and survived well in nature. The ethanol extract of plantlets from both in vivo & in vitro sources were prepared in soxhlet extractor and then concentrated to dryness under reduced pressure in rotary evaporator. Thus obtainedconcentrated extracts showed significant inhibitory activity against gram negative bacteria like Escherichia coli and Pseudomonas aeruginosa but no inhibition was found against gram positive bacteria. Further, these ethanol extracts were screened for in vitro percentage cytotoxicity at different time periods (24 h, 48 h and 72 h) of different dilutions. The in vivo plant extract inhibited the growth of EAC mouse cell lines in the range of 65, 66, 78, and 88% at 100, 50, 25 & 12.5μg mL-1 but at 72 h of treatment. In case of the extract of in vitro origin, the inhibition was found against EAC cell lines even at 48h. During spectrophotometric scanning, the extracts exhibited different maxima (ʎ) - four peaks in in vitro extracts as against single in in vivo preparation suggesting the possible change in the nature of ingredients during micropropagation through tissue culture techniques.

Keywords: Anti-cancer, Anti-microbial, EAC mouse cell, Tissue culture, Vernonia divergens.

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Authors: Koo-Yeon Kim, Eun-Hye Kim, Tae-Il Son

Abstract:

Epidermal Growth Factor (EGF, Mw=6,045) has been reported to have high efficiency of wound repair and anti-wrinkle effect. However, the half-life of EGF in the body is too short to exert the biological activity effectively when applied in free form. Growth Factors can be stabilized by immobilization with carbohydrates from thermal and proteolytic degradation. Low molecular weight chitosan (LMCS) and its derivate prepared by hydrogen peroxide has high solubility. LM6A6DC was successfully prepared as a reactive carbohydrate for the stabilization of EGF by the reactions of LMCS with alkalization, tosylation, azidation and reduction. The structure of LM6A6DC was confirmed by FT-IR, 1H NMR and elementary analysis. For enhancing the stability of free EGF, EGF was attached with LM6A6DC by using water-soluble carbodiimide. EGF-LM6A6DC conjugates did not show any cytotoxicity on the Normal Human Dermal Fibroblast (NHDF) 3T3 proliferation at least under 100 μg/ml. In the result, it was considered that LM6A6DC is suitable to immobilize of growth factor.

Keywords: Epidermal growth factor (EGF), low-molecular-weight chitosan, immobilization.

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Authors: Yahia M., Laanani I., Benbia S., Hachemi M., Massinissa Y.

Abstract:

Uterine and oviducal fluids are necessary for capacitation of the spermatozoa and early embryonic development. The aim of the present study was to determine the effects of estrous cycle phases (follicular and luteal) on some biological parameters (enzymes, electrolytes and total proteins) in uterine and oviducal secretions of ewes. Oviducal and uterine fluids were collected, diluted and centrifuged. According to our results, concentrations of GPT, G6PDH, total proteins, K and Na were significantly (P <0.05) higher at the luteal phase, however, the levels of aldolase, Mg, Ca and P were significantly (P <0.05) higher at the follicular phase in uterine secretions. While, only oviducal K and Ca were significantly (P<0.05) higher at the follicular phase. Our study revealed the existence of significant cyclic variations for some uterine and oviducal parameters which indicates the effect of ovarian hormones on the components of genital secretions.

Keywords: Biochemical parameters, estrous cycle, ewe, genital secretion.

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Authors: Mustafa M. Donma, Orkide Donma

Abstract:

Childhood obesity is an important clinical problem, because it may lead to chronic diseases during the adulthood period of the individual. Obesity is a metabolic disease associated with low-grade inflammation. The liver occurs at the center of metabolic pathways. Adropin, fibroblast growth factor-21 (FGF-21) and fetuin A are hepatokines. Due to the immense participation of the liver in glucose metabolism, these liver derived factors may be associated with insulin resistance (IR), which is a phenomenon discussed within the scope of obesity problems. The aim of this study is to determine the concentrations of adropin, FGF-21 and fetuin A in childhood obesity, to point out possible differences between the obesity groups and to investigate possible associations among these three hepatokines in obese and morbid obese children. A total of 132 children were included in the study. Two obese groups were constituted. The groups were matched in terms of mean±SD values of ages. Body mass index values of the obese and morbid obese groups were 25.0±3.5 kg/m2 and 29.8±5.7 kg/m2, respectively. Anthropometric measurements including waist circumference, hip circumference, head circumference, and neck circumference were recorded. Informed consent forms were taken from the parents of the participants and the Ethics Committee of the institution approved the study protocol. Blood samples were obtained after an overnight fasting. Routine biochemical tests including glucose- and lipid-related parameters were performed. Concentrations of the hepatokines (adropin, FGF-21, fetuin A) were determined by enzyme-linked immunosorbent assay. Insulin resistance indices such as homeostasis model assessment for IR (HOMA-IR), alanine transaminase-to aspartate transaminase ratio (ALT/AST), diagnostic obesity notation model assessment laboratory index, diagnostic obesity notation model assessment metabolic syndrome index as well as obesity indices such as diagnostic obesity notation model assessment-II index, and fat mass index were calculated using the previously derived formulas. Statistical evaluation of the study data as well as findings of the study were performed by SPSS for Windows. Statistical difference was accepted significant when p < 0.05. Statistically significant differences were found for insulin, triglyceride, high density lipoprotein cholesterol levels of the groups. A significant increase was observed for FGF-21 concentrations in the morbid obese group. Higher adropin and fetuin A concentrations were observed in the same group in comparison with the values detected in the obese group (p > 0.05). There was no statistically significant difference between the ALT/AST values of the groups. In all of the remaining IR and obesity indices, significantly increased values were calculated for morbid obese children. Significant correlations were detected between HOMA-IR and each of the hepatokines. The highest one was the association with fetuin A (r = 0.373, p = 0.001). In conclusion, increased levels observed in adropin, FGF-21 and fetuin A have shown that these hepatokines possess increasing potential going from the obese to morbid obese state. Out of the correlations found with IR index, the most affected hepatokine was fetuin A, the parameter possibly used as the indicator of the advanced obesity stage.

Keywords: adropin, fetuin A, fibroblast growth factor-21, insulin resistance, pediatric obesity

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