Reassessment of SST4 Somatostatin Receptor Expression Using SST4-eGFP Knockin Mice and the Novel Rabbit Monoclonal Anti-Human SST4 Antibody 7H49L61

Among the five somatostatin receptors (SST1–SST5), SST4 is the least characterized, which is in part due to the lack of specific monoclonal antibodies. We generated a knockin mouse model that expresses a carboxyl-terminal SST4-eGFP fusion protein. In addition, we extensively characterized the novel rabbit monoclonal anti-human SST4 antibody 7H49L61 using transfected cells and receptor-expressing tissues. 7H49L61 was then subjected to immunohistochemical staining of a series of formalin-fixed, paraffin-embedded normal and neoplastic human tissues. Characterization of SST4-eGFP mice revealed prominent SST4 expression in cortical pyramidal cells and trigeminal ganglion cells. In the human cortex, 7H49L61 disclosed a virtually identical staining pattern. Specificity of 7H49L61 was demonstrated by detection of a broad band migrating at 50–60 kDa in immunoblots. Tissue immunostaining was abolished by preadsorption of 7H49L61 with its immunizing peptide. In the subsequent immunohistochemical study, 7H49L61 yielded a predominant plasma membrane staining in adrenal cortex, exocrine pancreas, and placenta. SST4 was also found in glioblastomas, parathyroid adenomas, gastric and pancreatic adenocarcinomas, pheochromocytomas, and lymphomas. Altogether, we provide the first unequivocal localization of SST4 in normal and neoplastic human tissues. The monoclonal antibody 7H49L61 may also prove of great value for identifying SST4-expressing tumors during routine histopathological examinations.     

Impact of DICER1 and DROSHA on the Angiogenic Capacity of Human Endothelial Cells

RNAi-mediated knockdown of DICER1 and DROSHA, enzymes critically involved in miRNA biogenesis, has been postulated to affect the homeostasis and the angiogenic capacity of human endothelial cells. To re-evaluate this issue, we reduced the expression of DICER1 or DROSHA by RNAi-mediated knockdown and subsequently investigated the effect of these interventions on the angiogenic capacity of human umbilical vein endothelial cells (HUVEC) in vitro (proliferation, migration, tube formation, endothelial cell spheroid sprouting) and in a HUVEC xenograft assay in immune incompetent NSG^TM mice in vivo. In contrast to previous reports, neither knockdown of DICER1 nor knockdown of DROSHA profoundly affected migration or tube formation of HUVEC or the angiogenic capacity of HUVEC in vivo. Furthermore, knockdown of DICER1 and the combined knockdown of DICER1 and DROSHA tended to increase VEGF-induced BrdU incorporation and induced angiogenic sprouting from HUVEC spheroids. Consistent with these observations, global proteomic analyses showed that knockdown of DICER1 or DROSHA only moderately altered HUVEC protein expression profiles but additively reduced, for example, expression of the angiogenesis inhibitor thrombospondin-1. In conclusion, global reduction of miRNA biogenesis by knockdown of DICER1 or DROSHA does not inhibit the angiogenic capacity of HUVEC. Further studies are therefore needed to elucidate the influence of these enzymes in the context of human endothelial cell-related angiogenesis.     

Kidney Injury Caused by Preeclamptic Pregnancy Recovers Postpartum in a Transgenic Rat Model

Preeclampsia (PE) is characterized by the onset of hypertension (≥140/90 mmHg) and presence of proteinuria (>300 mg/L/24 h urine) or other maternal organ dysfunctions. During human PE, renal injuries have been observed. Some studies suggest that women with PE diagnosis have an increased risk to develop renal diseases later in life. However, in human studies PE as a single cause of this development cannot be investigated. Here, we aimed to investigate the effect of PE on postpartum renal damage in an established transgenic PE rat model. Female rats harboring the human-angiotensinogen gene develop a preeclamptic phenotype after mating with male rats harboring the human-renin gene, but are normotensive before and after pregnancy. During pregnancy PE rats developed mild tubular and glomerular changes assessed by histologic analysis, increased gene expression of renal damage markers such as kidney injury marker 1 and connective-tissue growth factor, and albuminuria compared to female wild-type rats (WT). However, four weeks postpartum, most PE-related renal pathologies were absent, including albuminuria and elevated biomarker expression. Only mild enlargement of the glomerular tuft could be detected. Overall, the glomerular and tubular function were affected during pregnancy in the transgenic PE rat. However, almost all these pathologies observed during PE recovered postpartum.     

Relaxation Pathways in Metallic Glasses

At temperatures below the glass transition temperature, physical properties of metallic glasses, such as density, viscosity, electrical resistivity or enthalpy, slowly evolve with time. This is the process of physical aging that occurs among all types of glasses and leads to structural changes at the microscopic level. Even though the relaxation pathways are ruled by thermodynamics as the glass attempts to re-attain thermodynamic equilibrium, they are steered by sluggish kinetics at the microscopic level. Understanding the structural and dynamic pathways of the relaxing glassy state is still one of the grand challenges in materials physics. We review some of the recent experimental advances made in understanding the nature of the relaxation phenomenon in metallic glasses and its implications to the macroscopic and microscopic properties changes of the relaxing glass.     

Analysis of heregulin symmetry by weighted evolutionary tracing

Heregulins are members of the protein family of EGF-like growthand differentiation factors. The primary cell-surface targetsof heregulins are heterodimers of the EGF-receptor homolog HER2with either HER3 or HER4. We used a weighted evolutionary traceanalysis to identify structural features that distinguish theEGF-like domain (hrg) of heregulins from other members of theEGF family. In this analysis, each amino acid sequence is weightedaccording to its uniqueness and the variability in each positionis assigned by an amino acid substitution matrix. Conservedresidues in heregulin that are variable in other EGF-like domainsare considered possible specificity-conferring residues. Thisanalysis identifies two clusters of residues at the foot ofthe boot-shaped hrg domain. The residues in one cluster arerecruited from the N-terminus; those in the other are from the-loop region and show a weak sequence similarity to the N-terminalresidues at the opposite side of the boot. The remaining residueswith high conservation scores distribute themselves into thesetwo distinct surfaces on hrg. This pseudo-twofold symmetry andthe presence of two distinct interfaces may reflect the preferenceof hrg for heterodimeric versus homodimeric HER complexes.     

Dense silicon carbonitride ceramics by pyrolysis of cross-linked and warm pressed polysilazane powders

This study reports on the pyrolysis and densifaction behavior of cross-linked poly(hydridomethylsilazane) powders. The influence of the cross-linking procedure such as temperature and annealing time of the polymer powders on the compaction behavior under cold and warm pressing conditions is discussed. The degree of cross-linking is determined by thermal mechanical analysis (TMA). In addition to particle sliding which is assumed to be the compaction mechanism obtained by cold-pressing, the polymer powder consolidates by plastic deformation applying warm-pressing. A continuous 3-dimensional polysilazane network is formed after a dwelling time under these conditions. Pyrolysis of the cross-linked and compacted polysilazane powder in argon at 1100°C gives crack-free amorphous silicon carbonitride Si_3+xC_x+yN₄ with compositions ranging from x=1·47 and y=0·88 for cold pressed samples to x=1·47 and y=1·86 for warm pressed materials. The residual open porosity is significantly reduced from 10–15 vol% in the cold pressed specimens to 1·3–5 vol% by the warm pressing procedure. The weight loss during pyrolysis between room temperature and 1300°C is about 5 wt% lower than that for cold pressed specimens. This result is explained by a reduced methane evolution during the polymer-to-ceramic conversion and is in accordance with the enhanced carbon content of the warm pressed material.     

A low-complexity low-spurs digital architecture for wideband PLL applications

This paper presents a non-ideality analysis of the wideband frequency synthesizer, and proposes a new digital architecture for signal modulation and quantization. Using error masking strategy, this method can offer smaller routing area and less power consumption without sacrificing any system performance. Simulation results confirm the theoretical predictions. Finally, the design guidelines are derived to fulfill the customized synthesizer specifications.