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RationaleThe mechanisms underlying atrial fibrillation (AF), the most common clinical arrhythmia, are poorly understood. Nucleoplasmic Ca2+ regulates gene expression, but the nature and significance of nuclear Ca2+-changes in AF are largely unknown.ObjectiveTo elucidate mechanisms by which AF alters atrial-cardiomyocyte nuclear Ca2+ ([Ca2+]Nuc) and CaMKII (Ca2+/calmodulin-dependent protein kinase-II)-related signaling.Methods and resultsAtrial cardiomyocytes were isolated from control and AF dogs (kept in AF by atrial tachypacing [600 bpm × 1 week]). [Ca2+]Nuc and cytosolic [Ca2+] ([Ca2+]Cyto) were recorded via confocal microscopy. Diastolic [Ca2+]Nuc was greater than [Ca2+]Cyto under control conditions, while resting [Ca2+]Nuc was similar to [Ca2+]Cyto; both diastolic and resting [Ca2+]Nuc increased with AF. IP3R (Inositol-trisphosphate receptor) stimulation produced larger [Ca2+]Nuc increases in AF versus control cardiomyocytes, and IP3R-blockade suppressed the AF-related [Ca2+]Nuc differences. AF upregulated nuclear protein expression of IP3R1 (IP3R-type 1) and of phosphorylated CaMKII (immunohistochemistry and immunoblot) while decreasing the nuclear/cytosolic expression ratio for HDAC4 (histone deacetylase type-4). Isolated atrial cardiomyocytes tachypaced at 3 Hz for 24 hours mimicked AF-type [Ca2+]Nuc changes and L-type calcium current decreases versus 1-Hz-paced cardiomyocytes; these changes were prevented by IP3R knockdown with short-interfering RNA directed against IP3R1. Nuclear/cytosolic HDAC4 expression ratio was decreased by 3-Hz pacing, while nuclear CaMKII phosphorylation was increased. Either CaMKII-inhibition (by autocamtide-2-related peptide) or IP3R-knockdown prevented the CaMKII-hyperphosphorylation and nuclear-to-cytosolic HDAC4 shift caused by 3-Hz pacing. In human atrial cardiomyocytes from AF patients, nuclear IP3R1-expression was significantly increased, with decreased nuclear/nonnuclear HDAC4 ratio. MicroRNA-26a was predicted to target ITPR1 (confirmed by luciferase assay) and was downregulated in AF atrial cardiomyocytes; microRNA-26a silencing reproduced AF-induced IP3R1 upregulation and nuclear diastolic Ca2+-loading.ConclusionsAF increases atrial-cardiomyocyte nucleoplasmic [Ca2+] by IP3R1-upregulation involving miR-26a, leading to enhanced IP3R1-CaMKII-HDAC4 signaling and L-type calcium current downregulation. Graphic Abstract: A graphic abstract is available for this article.

Surface modification of workpieces for cold forming processes is common in industrial practice. Specific surface topographies manufactured by means of shot peening form lubricant pockets, which provide a continuous lubricating film throughout the process. Furthermore, hydrostatic as well as hydrodynamic lubrication effects emerge, which further reduce the friction during the process and protect the tool from wear effects. These effects are well researched and applied in order to enable cold forming processes. In dry forming processes, no lubricants are used, and the positive lubrication effects do not appear. Surface modifications by shot peening or knurling change surface integrity in terms of hardness, residual stresses and geometry. The effects of the changed surface integrity on the forming process are not researched. In tribometer tests, a hard and rough surface texture lead to a lower frictional shear stress, whereas a rather smooth and hard surface texture leads to a lower punch force in an extrusion process. The mechanisms between surface textures and the smoothing and forming process are unknown. Furthermore, findings of the tribometer tests cannot be fully transferred to the extrusion process. As in tribometer tests a pin is used as tool and in extrusion a whole die, a discrepancy between the most suitable surface texture for a friction reduction results out of different flow conditions. Still, the tribometer tests give a first reference on a surface pretreament. The paper deals with an investigation of surface textures in order to distinguish characteristics of surface textures respectively a friction reduction. By means of indentation tests of the quenched and tempered steel 42CrMo4 the plastic flow in dependency of different surface integrities are researched. The results show which characteristics help to lower the punch force the most, so for further investigations an optimized surface modification is set up in order to enable a dry metal forming process.

An an iron-analogue of kinoshitalite occurs in the Broken Hill deposit in the Namaqualand Metamorphic Complex, South Africa. Like kinoshitalite and anandite, which are also found at Broken Hill, it is a brittle, baric, trioctahedral mica. Iron occurs in substantial substitution of magnesium. These characteristics indicate that barian micas of the Broken Hill area formed at around 670 degrees C and at around 4.5 kbar of pressure.

Abstract Background Oncolytic adenoviruses are promising agents for the multimodal treatment of cancer. However, tumor-selectivity is crucial for their applicability in patients. Recent studies by several groups demonstrated that oncolytic adenoviruses with tumor-/tissue-specific expression of the E1 and E4 genes, which are pivotal for adenoviral replication, have a specificity profile that is superior to viruses that solely target the expression of E1 or E4 genes. Presently the E1 and E4 regions are modified in a time consuming sequential fashion. Results Based on the widely used adenoviral cloning system AdEasy we generated a novel transfer vector that allows efficient and rapid generation of conditionally replication-competent adenovirus type 5 based vectors with the viral E1 and E4 genes under the transcriptional control of heterologous promoters. For insertion of the promoters of interest our transfer vector has two unique multiple cloning sites. Additionally, our shuttle plasmid allows encoding of a transgene within the E1A transcription unit. The modifications, including E1 mutations, are introduced into the adenoviral genome by a single homologous recombination step in Escherichia coli. Subsequently infectious viruses are rescued from plasmids. As a proof-of-concept we generated two conditionally replication-competent adenoviruses Ad.Ki•COX and Ad.COX•Ki with the promoters of the Ki-67 protein and the cyclooxygenase-2 (COX-2) driving E1 and E4 and vice versa. Conclusion We demonstrated with our cloning system efficient generation of double heterologous promoter controlled oncolytic adenoviral vectors by a single homologous recombination step in bacteria. The generated viruses showed preferential replication in tumor cells and in a subcutaneous HT-29 colon cancer xenograft model the viruses demonstrated significant oncolytic activity comparable with dl327.

Cold massive forming, particularly forward extrusion of steel, is characterized by high material utilization, product quality, and resource efficiency. Currently, enormous quantities of mostly ecologically harmful lubricants are required, leading to the demand for lubricant‐free cold forming. Furthermore, cost savings are achievable due to shorter lead times. However, the requirements for the tools cannot be met without the application of wear‐resistant and friction‐reducing hard coatings. The self‐lubricating hard coatings (Cr,Al)N + Mo:S and (Cr,Al)N + W:S with attuned material design deposited by physical vapor deposition (PVD) offer high potential for the realization of dry cold forming of steel. The process chain for the comprehensive characterization of the developed coating systems comprises analyses of coating and compound properties as well as two different model tests to determine the tribological behavior. One of the used tribometers represents an open tribological system, which is particularly suitable for simulating the loads occurring during cold massive forming of steel. The analyses reveal that with Mo and S modified coatings exhibit higher potential for ecologically sustainable cold forming due to their better mechanical properties and compound properties between the coatings and the cold work steel substrate compared with W and S modified coatings. [ABSTRACT FROM AUTHOR]

Background: Oncolytic adenoviruses are promising agents for the multimodal treatment of cancer. However, tumor-selectivity is crucial for their applicability in patients. Recent studies by several groups demonstrated that oncolytic adenoviruses with tumor-/tissue-specific expression of the E1 and E4 genes, which are pivotal for adenoviral replication, have a specificity profile that is superior to viruses that solely target the expression of E1 or E4 genes. Presently the E1 and E4 regions are modified in a time consuming sequential fashion. Results: Based on the widely used adenoviral cloning system AdEasy we generated a novel transfer vector that allows efficient and rapid generation of conditionally replication-competent adenovirus type 5 based vectors with the viral E1 and E4 genes under the transcriptional control of heterologous promoters. For insertion of the promoters of interest our transfer vector has two unique multiple cloning sites. Additionally, our shuttle plasmid allows encoding of a transgene within the E1A transcription unit. The modifications, including E1 mutations, are introduced into the adenoviral genome by a single homologous recombination step in Escherichia coli. Subsequently infectious viruses are rescued from plasmids. As a proof-of-concept we generated two conditionally replication-competent adenoviruses Ad.Ki•COX and Ad.COX•Ki with the promoters of the Ki-67 protein and the cyclooxygenase-2 (COX-2) driving E1 and E4 and vice versa. Conclusion: We demonstrated with our cloning system efficient generation of double heterologous promoter controlled oncolytic adenoviral vectors by a single homologous recombination step in bacteria. The generated viruses showed preferential replication in tumor cells and in a subcutaneous HT-29 colon cancer xenograft model the viruses demonstrated significant oncolytic activity comparable with dl327. [ABSTRACT FROM AUTHOR]