Comparison of Anti-Shadoo Antibodies – Where is the Endogenous Shadoo protein?

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.

Segmentation of Cardiac Images by the Force Field Driven Speed Term

The class of geometric deformable models, so-called level sets, has brought tremendous impact to medical imagery. In this paper we present yet another application of level sets to medical imaging. The method we give here will in a way modify the speed term in the standard level sets equation of motion. To do so we build a potential based on the distance and the gradient of the image we study. In turn the potential gives rise to the force field: F~F(x, y) = P ∀(p,q)∈I ((x, y) - (p, q)) |ÔêçI(p,q)| |(x,y)-(p,q)| 2 . The direction and intensity of the force field at each point will determine the direction of the contour-s evolution. The images we used to test our method were produced by the Univesit'e de Sherbrooke-s PET scanners.

The Optimal Design for Grip Force of Material Handling

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.

Screening and Evaluation of in vivo and in vitro Generated Insulin Plant (Vernonia divergens) for Antimicrobial and Anticancer Activities

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.

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

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.

A Laser Point Interaction System Integrating Mouse Functions

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.

Magnetic Field Based Near Surface Haptic and Pointing Interface

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.

Interactive PTZ Camera Control System Using Wii Remote and Infrared Sensor Bar

This paper proposes an alternative control mechanism for an interactive Pan/Tilt/Zoom (PTZ) camera control system. Instead of using a mouse or a joystick, the proposed mechanism utilizes a Nintendo Wii remote and infrared (IR) sensor bar. The Wii remote has buttons that allows the user to control the movement of a PTZ camera through Bluetooth connectivity. In addition, the Wii remote has a built-in motion sensor that allows the user to give control signals to the PTZ camera through pitch and roll movement. A stationary IR sensor bar, placed at some distance away opposite the Wii remote, enables the detection of yaw movement. In addition, the Wii remote-s built-in IR camera has the ability to detect its spatial position, and thus generates a control signal when the user moves the Wii remote. Some experiments are carried out and their performances are compared with an industry-standard PTZ joystick.

Effect of White Kwao Extract (Pueraria mirifica) on in vitro Development and Implantation Rate of Mouse Embryo

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

Embryo Transfer as an Assisted Reproductive Technology in Farm Animals

Various assisted reproductive techniques have been developed and refined to obtain a large number of offspring from genetically superior animals or obtain offspring from infertile (or subfertile) animals. The embryo transfer is one assisted reproductive technique developed well, aimed at increased productivity of selected females, disease control, importation and exportation of livestock, rapid screening of AI sires for genetically recessive characteristics, treatment or circumvention of certain types of infertility. Embryo transfer also is a useful research tool for evaluating fetal and maternal interactions. This technique has been applied to nearly every species of domestic animal and many species of wildlife and exotic animals, including humans and non-human primates. The successful of embryo transfers have been limited to within-animal, homologous replacement of the embryos. There are several examples of interspecific and intergeneric embryo transfers in which embryos implanted but did not develop to term: sheep and goat, mouse and rat. An immunological rejections and placental incompatibility between the embryo and the surrogate mother appear to restrict interspecific embryo transfer/interspecific pregnancy. Recently, preimplantation embryo manipulation procedures have been applied, such as technique of inner cell mass transfer. This technique will possible to overcome the reproductive barrier interspecific embryo transfer/interspecific pregnancy, if there is a protective mechanism which prevents recognition of the foreign fetus by the mother of the other species

The Role of Immunogenic Adhesin Vibrio alginolyticus 49 k Da to Molecule Expression of Major Histocompatibility Complex on Receptors of Humpback Grouper Cromileptes altivelis

The purpose of research was to know the role of immunogenic protein of 49 kDa from V.alginolyticus which capable to initiate molecule expression of MHC Class II in receptor of Cromileptes altivelis. The method used was in vivo experimental research through testing of immunogenic protein 49 kDa from V.alginolyticus at Cromileptes altivelis (size of 250 - 300 grams) using 3 times booster by injecting an immunogenic protein in a intramuscular manner. Response of expressed MHC molecule was shown using immunocytochemistry method and SEM. Results indicated that adhesin V.alginolyticus 49 kDa which have immunogenic character could trigger expression of MHC class II on receptor of grouper and has been proven by staining using immunocytochemistry and SEM with labeling using antibody anti MHC (anti mouse). This visible expression based on binding between epitopes antigen and antibody anti MHC in the receptor. Using immunocytochemistry, intracellular response of MHC to in vivo induction of immunogenic adhesin from V.alginolyticus was shown.

Mouse Pointer Tracking with Eyes

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.

Retina Based Mouse Control (RBMC)

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.

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

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.

The First Prevalence Report of Direct Identification and Differentiation of B. abortus and B. melitensis using Real Time PCR in House Mouse of Iran

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.

Metabolic Analysis of Fibroblast Conditioned Media and Comparison with Theoretical Modeling

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.

A Novel Nucleus-Based Classifier for Discrimination of Osteoclasts and Mesenchymal Precursor Cells in Mouse Bone Marrow Cultures

Bone remodeling occurs by the balanced action of bone resorbing osteoclasts (OC) and bone-building osteoblasts. Increased bone resorption by excessive OC activity contributes to malignant and non-malignant diseases including osteoporosis. To study OC differentiation and function, OC formed in in vitro cultures are currently counted manually, a tedious procedure which is prone to inter-observer differences. Aiming for an automated OC-quantification system, classification of OC and precursor cells was done on fluorescence microscope images based on the distinct appearance of fluorescent nuclei. Following ellipse fitting to nuclei, a combination of eight features enabled clustering of OC and precursor cell nuclei. After evaluating different machine-learning techniques, LOGREG achieved 74% correctly classified OC and precursor cell nuclei, outperforming human experts (best expert: 55%). In combination with the automated detection of total cell areas, this system allows to measure various cell parameters and most importantly to quantify proteins involved in osteoclastogenesis.