Beta-COP is essential for biosynthetic membrane transport from the endoplasmic reticulum to the Golgi complex in vivo

Experimental glomerulonephritis was induced in rats to investigate the consequence of the antigen-antibody interaction on the surface of glomerular endothelial cells (GENs). A lectin, Lens culinaris hemoagglutinin (LCH), was first planted in the left kidney by isolated perfusion of a left kidney, and then the circulation was reestablished. Rabbit anti-LCH antibodies were injected from the tail vein 3 minutes after the recirculation of the left kidney, and acute glomerulonephritis ensued. Fifteen minutes after the injection, rabbit immunoglobulin G (IgG), rat C3, and LCH were observed exclusively on the surface of GENs. Accumulation of platelets was prominent. Three hours later, the immune deposits were seen in the subendothelial space, and the polymorphonuclear cells were the dominant infiltrate in the glomeruli. Up to the seventh day, immune deposits were seen in the subendothelial space, and the widening of this area was increasingly observed. Fourteen days later, immune deposits containing rat IgG were observed in the subepithelial area, but they were only occasionally seen in the subendothelial space and in the mesangial area. No crescent formation was seen at day 14, but the mesangial area was expanded, with an increased number of cells. The number of nuclei in the cross-section of a glomerulus increased after the induction of glomerulonephritis, but the number of leukocyte common antigen-positive cells (infiltrating cells) decreased gradually from day 4 to day 14. The staining of Thy-1.1, a marker of mesangial cell, was markedly enlarged in the glomerulus at day 14. These data suggest that mesangial proliferative glomerulonephritis can be induced by the antigen-antibody interaction on the surface of GENs.     

Several ways to produce porous carbon monoliths by template assisted routes

Several ways to produce microporous and mesoporous carbon monoliths by template assisted synthesis routes are described. Pressed zeolite (HY) and silica powder tablets have been used as templates. Sucrose acts as a carbon precursor and as a binder between the carbon particles as well. In the case of zeolite template, the carbon porosity could be increased by additional loading of the template with ethene. Due to the balance between mechanic stability and porosity of the carbon monolith, an optimal amount of sucrose is required. The final products were characterized by low temperature nitrogen adsorption.     

Extended bisection method for parameter identification of the transient heat conduction equation for thermo-elastic deformations during drilling

A new interdisciplinary approach is discribed to identifying unknown parameters using an extended version of the known interval bisection method. This developed method is based on the use of finite elements for calibrating the simulation calculation. The resulting thermo-elastic deformations which occur in drilling processes with impaired cooling lubrication are to be used as correction values for tool positioning in the NC control. Based on the strong impact on workpiece temperature of machining, a simulation approach is presented for calculating the temperature fields and their thermo-elastic consequences. In addition, methods are presented to correct these effects. This paper particularly deals with the temperature fields of drilling operations. Special attention is paid to the technique employed for iterative numerical determination of the unknown heat flux η _w and heat transfer coefficient \(\bar {\gamma }\) values. Finally, the data obtained from experiments are compared with those achieved by numerical simulation in order to verify the efficiency of simulation and determination of parameters.     

The Functional Characterization of GCaMP3.0 Variants Specifically Targeted to Subcellular Domains

Calcium (Ca²⁺) ions play a pivotal role in physiology and cellular signaling. The intracellular Ca²⁺ concentration ([Ca²⁺]_i) is about three orders of magnitude lower than the extracellular concentration, resulting in a steep transmembrane concentration gradient. Thus, the spatial and the temporal dynamics of [Ca²⁺]_i are ideally suited to modulate Ca²⁺-mediated cellular responses to external signals. A variety of highly sophisticated methods have been developed to gain insight into cellular Ca²⁺ dynamics. In addition to electrophysiological measurements and the application of synthetic dyes that change their fluorescent properties upon interaction with Ca²⁺, the introduction and the ongoing development of genetically encoded Ca²⁺ indicators (GECI) opened a new era to study Ca²⁺-driven processes in living cells and organisms. Here, we have focused on one well-established GECI, i.e., GCaMP3.0. We have systematically modified the protein with sequence motifs, allowing localization of the sensor in the nucleus, in the mitochondrial matrix, at the mitochondrial outer membrane, and at the plasma membrane. The individual variants and a cytosolic version of GCaMP3.0 were overexpressed and purified from E. coli cells to study their biophysical properties in solution. All versions were examined to monitor Ca²⁺ signaling in stably transfected cell lines and in primary cortical neurons transduced with recombinant Adeno-associated viruses (rAAV). In this comparative study, we provide evidence for a robust approach to reliably trace Ca²⁺ signals at the (sub)-cellular level with pronounced temporal resolution.     

„Ideale“ Lötverbindung und „optimale“ Lötverbindung

If due to the market analysis a company makes the decision for the production of a new product by application of soldering and special the soldering with temporarily liquid solder, this one is then made specifically after that the company specific construction and production preparation. Either the decision for soldering is technically and/or economically well‐founded or she is even unavoidable by so‐called mixed material connections in the assemblies. This hybrid design finds application increasing today. In the contribution on hand the analysis and assessment of the soldering theory and practice is carried out under the point of view of the possible production of “ideal soldered connections” and “optimal soldered connections”.     

Overexpression of microRNAs miR-25-3p,miR-185-5p and miR-132-3p in Late Onset Fetal Growth Restriction,Validation of Results and Study of the Biochemical Pathways Involved

In a prospective study, 48 fetuses were evaluated with Doppler ultrasound after 34 weeks and classified, according to the cerebroplacental ratio (CPR) and estimated fetal weight (EFW), into fetuses with normal growth and fetuses with late-onset fetal growth restriction (LO-FGR). Overexpression of miRNAs from neonatal cord blood belonging to LO-FGR fetuses, was validated by real-time PCR. In addition, functional characterization of overexpressed miRNAs was performed by analyzing overrepresented pathways, gene ontologies, and prioritization of synergistically working miRNAs. Three miRNAs: miR-25-3p, miR-185-5p and miR-132-3p, were significantly overexpressed in cord blood of LO-FGR fetuses. Pathway and gene ontology analysis revealed over-representation of certain molecular pathways associated with cardiac development and neuron death. In addition, prioritization of synergistically working miRNAs highlighted the importance of miR-185-5p and miR-25-3p in cholesterol efflux and starvation responses associated with LO-FGR phenotypes. Evaluation of miR-25-3p; miR-132-3p and miR-185-5p might serve as molecular biomarkers for the diagnosis and management of LO-FGR; improving the understanding of its influence on adult disease.     

Peanut varieties with reduced Ara h 1 content indicating no reduced allergenicity

Peanut allergy is a major cause of food‐induced severe anaphylactic reactions. To date, no medical care is available to prevent and treat peanut allergy and therefore hypoallergenic peanut varieties are of considerable health political and economic interest. Major allergens that induce IgE‐responses in peanut‐sensitive patients are Ara h 1, Ara h 2 and Ara h 3/4. In order to identify hypoallergenic peanuts, commercially locally available peanut varieties were screened for their allergen content. Ara h 1‐deficient peanuts from Southeast Asia were identified by SDS‐PAGE, immunoblotting, inhibition assays and ELISA. 2‐D PAGE analyses demonstrated the different compositions of the tested extracts and revealed a number of variations of the allergen patterns of peanuts from different varieties. Mediator release experiments of these peanut extracts demonstrated similar allergenicities as compared with standard peanut extract. These results indicate that the allergenicity of peanuts with reduced Ara h 1 content might be compensated by the other allergens, and thus do not necessarily cause a reduction of allergenicity.     

Characterization of an Invertebrate-Type Dopamine Receptor of the American Cockroach,Periplaneta americana

We have isolated a cDNA coding for a putative invertebrate-type dopamine receptor (Peadop2) from P. americana brain by using a PCR-based strategy. The mRNA is present in samples from brain and salivary glands. We analyzed the distribution of the PeaDOP2 receptor protein with specific affinity-purified polyclonal antibodies. On Western blots, PeaDOP2 was detected in protein samples from brain, subesophageal ganglion, thoracic ganglia, and salivary glands. In immunocytochemical experiments, we detected PeaDOP2 in neurons with their somata being located at the anterior edge of the medulla bilaterally innervating the optic lobes and projecting to the ventro-lateral protocerebrum. In order to determine the functional and pharmacological properties of the cloned receptor, we generated a cell line constitutively expressing PeaDOP2. Activation of PeaDOP2-expressing cells with dopamine induced an increase in intracellular cAMP. In contrast, a C-terminally truncated splice variant of this receptor did not exhibit any functional property by itself. The molecular and pharmacological characterization of the first dopamine receptor from P. americana provides the basis for forthcoming studies focusing on the significance of the dopaminergic system in cockroach behavior and physiology.     

Quantification and localization of the liquid zone of partially remelted M2 tool steel using X-ray microtomography and scanning electron microscopy

Thixoforming of steels poses challenges due to the high temperatures involved and the lack of understanding of thermomechanical behavior. The volume fractions of the liquid and solid phases in the semi-solid state are the most important parameters for such a forming process, as they affect the viscosity and hence the flow behavior of the material. Two-dimensional observations might not always be sufficient, as the size distribution and the connectivity of phases cannot be obtained from associated measurements, which can only be determined by three-dimensional (3-D) investigation. This paper presents the first application of high-energy X-ray microtomography to the microstructure of steel in the semi-solid state. The microstructure of M2 high-speed tool steel was studied in both as-received and heated-and-quenched states. From the reconstructed images, 3-D information could be obtained and was compared with scanning electron microscopy and energy dispersive spectrometry observations. The volume fraction and the location of liquid phase in the semi-solid state were determined in particular, and the continuous solid skeleton was investigated.