Application of Staining Intensity Correlation Analysis to Visualize Protein Colocalizationat a Cellular Level

Mutations of the telomeric copy of the survival motor neuron 1 (SMN1) gene cause spinal muscular atrophy. A deletion of the Eef1a2 gene leads to lower motor neuron degeneration in wasted mice. Indirect evidences have been shown that the eEF1A protein family may interact with SMN, and our previous study showed that abnormalities of neuromuscular junctions in wasted mice were similar to those of Smn mutant mice. To determine potential colocalization between SMN and tissue-specific translation elongation factor 1A2 (eEF1A2), an immunochemical analysis of HeLa cells transfected with the plasmid pcDNA3.1(+)C-hEEF1A2- myc and a new quantitative test of colocalization by intensity correlation analysis (ICA) was used to explore the association of SMN and eEF1A2. Here the results showed that eEF1A2 redistributed from the cytoplasm to the nucleus in response to serum and epidermal growth factor. In the cytoplasm, compelling evidence showed that staining for myc-tagged eEF1A2 varied in synchrony with that for SMN, consistent with the formation of a SMN-eEF1A2 complex in the cytoplasm of HeLa cells. These findings suggest that eEF1A2 may colocalize with SMN in the cytoplasm and may be a component of the SMN complex. However, the limitation of the ICA method is an inability to resolve colocalization in components of small organelles such as the nucleus.

Molecular Characterization of Free Radicals Decomposing Genes on Plant Developmental Stages

Biochemical and molecular analysis of some antioxidant enzyme genes revealed different level of gene expression on oilseed (Brassica napus). For molecular and biochemical analysis, leaf tissues were harvested from plants at eight different developmental stages, from young to senescence. The levels of total protein and chlorophyll were increased during maturity stages of plant, while these were decreased during the last stages of plant growth. Structural analysis (nucleotide and deduced amino acid sequence, and phylogenic tree) of a complementary DNA revealed a high level of similarity for a family of Catalase genes. The expression of the gene encoded by different Catalase isoforms was assessed during different plant growth phase. No significant difference between samples was observed, when Catalase activity was statistically analyzed at different developmental stages. EST analysis exhibited different transcripts levels for a number of other relevant antioxidant genes (different isoforms of SOD and glutathione). The high level of transcription of these genes at senescence stages was indicated that these genes are senescenceinduced genes.

An Assessment of the Small Hydropower Potential of Sisakht Region of Yasuj

Energy generated by the force of water in hydropower can provide a more sustainable, non-polluting alternative to fossil fuels, along with other renewable sources of energy, such as wind, solar and tidal power, bio energy and geothermal energy. Small scale hydroelectricity in Iran is well suited for “off-grid" rural electricity applications, while other renewable energy sources, such as wind, solar and biomass, can be beneficially used as fuel for pumping groundwater for drinking and small scale irrigation in remote rural areas or small villages. Small Hydro Power plants in Iran have very low operating and maintenance costs because they consume no fossil or nuclear fuel and do not involve high temperature processes. The equipment is relatively simple to operate and maintain. Hydropower equipment can adjust rapidly to load changes. The extended equipment life provides significant economic advantages. Some hydroelectric plants installed 100 years ago still operate reliably. The Polkolo river is located on Karun basin at southwest of Iran. Situation and conditions of Polkolo river are evaluated for construction of small hydropower in this article. The topographical conditions and the existence of permanent water from springs provide the suitability to install hydroelectric power plants on the river Polkolo. The cascade plant consists of 9 power plants connected with each other and is having the total head as 1100m and discharge about 2.5cubic meter per second. The annual production of energy is 105.5 million kwh.

Debye Layer Confinement of Nucleons in Nuclei by Laser Ablated Plasma

Following the laser ablation studies leading to a theory of nuclei confinement by a Debye layer mechanism, we present here numerical evaluations for the known stable nuclei where the Coulomb repulsion is included as a rather minor component especially for lager nuclei. In this research paper the required physical conditions for the formation and stability of nuclei particularly endothermic nuclei with mass number greater than to which is an open astrophysical question have been investigated. Using the Debye layer mechanism, nuclear surface energy, Fermi energy and coulomb repulsion energy it is possible to find conditions under which the process of nucleation is permitted in early universe. Our numerical calculations indicate that about 200 second after the big bang at temperature of about 100 KeV and subrelativistic region with nucleon density nearly equal to normal nuclear density namely, 10cm all endothermic and exothermic nuclei have been formed.

Immune Responce in Mice Immunized with Live Cold-Adapted Influenza Vaccine in Combination with Chitosan-Based Adjuvants

An influence of intranasal combined injection of live cold-adapted influenza vaccine with chitosan derivatives as adjuvants on the subpopulation structure of mononuclear leukocytes of mouse spleen which reflects the orientation of the immune response was studied. It is found that the inclusion of chitosan preparations promotes activation of cellular-level of immune response.

Agrowaste: Phytosterol from Durian Seed

Presence of phytosterol compound in Durian seed (Durio zibethinus) or known as King of fruits has been discovered from screening work using reagent test. Further analysis work has been carried out using mass spectrometer in order to support the priliminary finding. Isolation and purification of the major phytosterol has been carried out using an open column chromatography. The separation was monitored using thin layer chromatography (TLC). Major isolated compounds and purified phytosterol were identified using mass spectrometer and nuclear magnetic resonance (NMR). This novel finding could promote utilization of durian seeds as a functional ingredient in food products through production of standardized extract based on phytosterol content.

Calculation of Masses and Magnetic Moment of the Nucleon using the MIT Bag Model

The bag radius of the nucleon can be determined by MIT bag model based on electric and magnetic form factors of the nucleon. Also we determined the masses and magnetic moment of the nucleon with MIT bag model, using bag radius and compared with other results, suggests a suitable compatibility.

SDS-induced Serine Protease Activity of an Antiviral Red Fluorescent Protein

A rare phenomenon of SDS-induced activation of a latent protease activity associated with the purified silkworm excretory red fluorescent protein (SE-RFP) was noticed. SE-RFP aliquots incubated with SDS for different time intervals indicated that the protein undergoes an obligatory breakdown into a number of subunits which exhibit autoproteolytic (acting upon themselves) and/or heteroproteolytic (acting on other proteins) activities. A strong serine protease activity of SE-RFP subunits on Bombyx mori nucleopolyhedrovirus (BmNPV) polyhedral protein was detected by zymography technique. A complete inhibition of BmNPV infection to silkworms was observed by the oral administration assay of the SE-RFP. Here, it is proposed that the SE-RFP prevents the initial infection of BmNPV to silkworms by obliterating the polyhedral protein. This is the first report on a silkworm red fluorescent protein that exhibits a protease activity on exposure to SDS. The present studies would help in understanding the antiviral mechanism of silkworm red fluorescent proteins.

Development and Evaluation of a Dynamic Cardiac Phantom for use in Nuclear Medicine

The aim of this study was to develop a dynamic cardiac phantom for quality control in myocardial scintigraphy. The dynamic heart phantom constructed only contained the left ventricle, made of elastic material (latex), comprising two cavities: one internal and one external. The data showed a non-significant variation in the values of left ventricular ejection fraction (LVEF) obtained by varying the heart rate. It was also possible to evaluate the ejection fraction (LVEF) through different arrays of image acquisition and to perform an intercomparison of LVEF by two different scintillation cameras. The results of the quality control tests were satisfactory, showing that they can be used as parameters in future assessments. The new dynamic heart phantom was demonstrated to be effective for use in LVEF measurements. Therefore, the new heart simulator is useful for the quality control of scintigraphic cameras.

Density Functional Calculations of 27Al, 11B,and 14N and NQR Parameters in the (6, 0) BN_AlN Nanotube Junction

Density functional theory (DFT) calculations were performed to calculate aluminum-27, boron-11, and nitrogen-14 quadrupole coupling constant (CQ) in the representative considered model of (6, 0) boron nitride-aluminum nitride nanotube junction (BN-AlNNT) for the first time. To this aim, 1.3 nm length of BNAlN consisting of 18 Al, 18 B, and 36 N atoms was selected where the end atoms capped by hydrogen atoms. The calculated CQ values for optimized BN-AlNNT system reveal different electrostatic environment in the mentioned system. The calculations were performed using the Gaussian 98 package of program.

Influence of Artificial Roughness on Heat Transfer in the Rotating Flow

The results of an experimental study of the process of convective and boiling heat transfer in the vessel with stirrer for smooth and rough ring-shaped pipes are presented. It is established that creation of two-dimensional artificial roughness on the heated surface causes the essential (~100%) intensification of convective heat transfer. In case of boiling the influence of roughness appears on the initial stage of boiling and in case of fully developed nucleate boiling there was no intensification of heat transfer. The similitude equation for calculating convective heat transfer coefficient, which generalizes well experimental data both for the smooth and the rough surfaces is proposed.

Shell Closures in Exotic Nuclei

Inspired by the recent experiments [1]-[3] indicating unusual doubly magic nucleus 24O which lies just at the neutron drip-line and encouraged by the success of our relativistic mean-field (RMF) plus state dependent BCS approach for the description of the ground state properties of the drip-line nuclei [23]-[27], we have further employed this approach, across the entire periodic table, to explore the unusual shell closures in exotic nuclei. In our RMF+BCS approach the single particle continuum corresponding to the RMF is replaced by a set of discrete positive energy states for the calculations of pairing energy. Detailed analysis of the single particle spectrum, pairing energies and densities of the nuclei predict the unusual proton shell closures at Z = 6, 14, 16, 34, and unusual neutron shell closures at N = 6, 14, 16, 34, 40, 70, 112.

Multichannel Image Mosaicing of Stem Cells

Image mosaicing techniques are usually employed to offer researchers a wider field of view of microscopic image of biological samples. a mosaic is commonly achieved using automated microscopes and often with one “color" channel, whether it refers to natural or fluorescent analysis. In this work we present a method to achieve three subsequent mosaics of the same part of a stem cell culture analyzed in phase contrast and in fluorescence, with a common non-automated inverted microscope. The mosaics obtained are then merged together to mark, in the original contrast phase images, nuclei and cytoplasm of the cells referring to a mosaic of the culture, rather than to single images. The experiments carried out prove the effectiveness of our approach with cultures of cells stained with calcein (green/cytoplasm and nuclei) and hoechst (blue/nuclei) probes.

Multi-agent On-line Monitor for the Safety of Critical Systems

Operational safety of critical systems, such as nuclear power plants, industrial chemical processes and means of transportation, is a major concern for system engineers and operators. A means to assure that is on-line safety monitors that deliver three safety tasks; fault detection and diagnosis, alarm annunciation and fault controlling. While current monitors deliver these tasks, benefits and limitations in their approaches have at the same time been highlighted. Drawing from those benefits, this paper develops a distributed monitor based on semi-independent agents, i.e. a multiagent system, and monitoring knowledge derived from a safety assessment model of the monitored system. Agents are deployed hierarchically and provided with knowledge portions and collaboration protocols to reason and integrate over the operational conditions of the components of the monitored system. The monitor aims to address limitations arising from the large-scale, complicated behaviour and distributed nature of monitored systems and deliver the aforementioned three monitoring tasks effectively.

Developments for ''Virtual'' Monitoring and Process Simulation of the Cryogenic Pilot Plant

The implementation of the new software and hardware-s technologies for tritium processing nuclear plants, and especially those with an experimental character or of new technology developments shows a coefficient of complexity due to issues raised by the implementation of the performing instrumentation and equipment into a unitary monitoring system of the nuclear technological process of tritium removal. Keeping the system-s flexibility is a demand of the nuclear experimental plants for which the change of configuration, process and parameters is something usual. The big amount of data that needs to be processed stored and accessed for real time simulation and optimization demands the achievement of the virtual technologic platform where the data acquiring, control and analysis systems of the technological process can be integrated with a developed technological monitoring system. Thus, integrated computing and monitoring systems needed for the supervising of the technological process will be executed, to be continued with the execution of optimization system, by choosing new and performed methods corresponding to the technological processes within the tritium removal processing nuclear plants. The developing software applications is executed with the support of the program packages dedicated to industrial processes and they will include acquisition and monitoring sub-modules, named “virtually" as well as the storage sub-module of the process data later required for the software of optimization and simulation of the technological process for tritium removal. The system plays and important role in the environment protection and durable development through new technologies, that is – the reduction of and fight against industrial accidents in the case of tritium processing nuclear plants. Research for monitoring optimisation of nuclear processes is also a major driving force for economic and social development.

A Novel Prediction Method for Tag SNP Selection using Genetic Algorithm based on KNN

Single nucleotide polymorphisms (SNPs) hold much promise as a basis for disease-gene association. However, research is limited by the cost of genotyping the tremendous number of SNPs. Therefore, it is important to identify a small subset of informative SNPs, the so-called tag SNPs. This subset consists of selected SNPs of the genotypes, and accurately represents the rest of the SNPs. Furthermore, an effective evaluation method is needed to evaluate prediction accuracy of a set of tag SNPs. In this paper, a genetic algorithm (GA) is applied to tag SNP problems, and the K-nearest neighbor (K-NN) serves as a prediction method of tag SNP selection. The experimental data used was taken from the HapMap project; it consists of genotype data rather than haplotype data. The proposed method consistently identified tag SNPs with considerably better prediction accuracy than methods from the literature. At the same time, the number of tag SNPs identified was smaller than the number of tag SNPs in the other methods. The run time of the proposed method was much shorter than the run time of the SVM/STSA method when the same accuracy was reached.

Principal Type of Water Responsible for Damage of Concrete Repeated Freeze-Thaw Cycles

The first and basic cause of the failure of concrete is repeated freezing (thawing) of moisture contained in the pores, microcracks, and cavities of the concrete. On transition to ice, water existing in the free state in cracks increases in volume, expanding the recess in which freezing occurs. A reduction in strength below the initial value is to be expected and further cycle of freezing and thawing have a further marked effect. By using some experimental parameters like nuclear magnetic resonance variation (NMR), enthalpy-temperature (or heat capacity) variation, we can resolve between the various water states and their effect on concrete properties during cooling through the freezing transition temperature range. The main objective of this paper is to describe the principal type of water responsible for the reduction in strength and structural damage (frost damage) of concrete following repeated freeze –thaw cycles. Some experimental work was carried out at the institute of cryogenics to determine what happens to water in concrete during the freezing transition. 

Role of Oxidative DNA Damage in Pathogenesis of Diabetic Neuropathy

Oxidative stress is considered to be the cause for onset and the progression of type 2 diabetes mellitus (T2DM) and complications including neuropathy. It is a deleterious process that can be an important mediator of damage to cell structures: protein, lipids and DNA. Data suggest that in patients with diabetes and diabetic neuropathy DNA repair is impaired, which prevents effective removal of lesions. Objective: The aim of our study was to evaluate the association of the hOGG1 (326 Ser/Cys) and XRCC1 (194 Arg/Trp, 399 Arg/Gln) gene polymorphisms whose protein is involved in the BER pathway with DNA repair efficiency in patients with diabetes type 2 and diabetic neuropathy compared to the healthy subjects. Genotypes were determined by PCR-RFLP analysis in 385 subjects, including 117 with type 2 diabetes, 56 with diabetic neuropathy and 212 with normal glucose metabolism. The polymorphisms studied include codon 326 of hOGG1 and 194, 399 of XRCC1 in the base excision repair (BER) genes. Comet assay was carried out using peripheral blood lymphocytes from the patients and controls. This test enabled the evaluation of DNA damage in cells exposed to hydrogen peroxide alone and in the combination with the endonuclease III (Nth). The results of the analysis of polymorphism were statistically examination by calculating the odds ratio (OR) and their 95% confidence intervals (95% CI) using the ¤ç2-tests. Our data indicate that patients with diabetes mellitus type 2 (including those with neuropathy) had higher frequencies of the XRCC1 399Arg/Gln polymorphism in homozygote (GG) (OR: 1.85 [95% CI: 1.07-3.22], P=0.3) and also increased frequency of 399Gln (G) allele (OR: 1.38 [95% CI: 1.03-1.83], P=0.3). No relation to other polymorphisms with increased risk of diabetes or diabetic neuropathy. In T2DM patients complicated by neuropathy, there was less efficient repair of oxidative DNA damage induced by hydrogen peroxide in both the presence and absence of the Nth enzyme. The results of our study suggest that the XRCC1 399 Arg/Gln polymorphism is a significant risk factor of T2DM in Polish population. Obtained data suggest a decreased efficiency of DNA repair in cells from patients with diabetes and neuropathy may be associated with oxidative stress. Additionally, patients with neuropathy are characterized by even greater sensitivity to oxidative damage than patients with diabetes, which suggests participation of free radicals in the pathogenesis of neuropathy.

Study of Peptide Fragment of Alpha-Fetoprotein as a Radionuclide Vehicle

Alpfa-fetoprotein and its fragments may be an important vehicle for targeted delivery of radionuclides to the tumor. We investigated the effect of conditions on the labeling of biologically active synthetic peptide based on the (F-afp) with technetium-99m. The influence of the nature of the buffer solution, pH, concentration of reductant, concentration of the peptide and the reaction temperature on the yield of labeling was examined. As a result, the following optimal conditions for labeling of (F-afp) are found: pH 8.5 (phosphate and bicarbonate buffers) and pH from 1.7 to 7.0 (citrate buffer). The reaction proceeds with sufficient yield at room temperature for 30 min at the concentration of SnCl2 and (Fafp) (F-afp) is to be less than 10 mkg/ml and 25 mkg/ml, respectively. Investigations of the test drug accumulation in the tumor cells of human breast cancer were carried out. Results can be assumed that the in vivo study of the (F-afp) in experimental tumor lesions will show concentrations sufficient for imaging these lesions by SPECT.

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.