Johanson-Blizzard syndrome (OMIM 243800) is an autosomal recessive disorder that includes congenital exocrine pancreatic insufficiency, multiple malformations such as nasal wing aplasia, and frequent ...mental retardation. We mapped the disease-associated locus to chromosome 15q14-21.1 and identified mutations, mostly truncating ones, in the gene UBR1 in 12 unrelated families with Johanson-Blizzard syndrome. UBR1 encodes one of at least four functionally overlapping E3 ubiquitin ligases of the N-end rule pathway, a conserved proteolytic system whose substrates include proteins with destabilizing N-terminal residues. Pancreas of individuals with Johanson-Blizzard syndrome did not express UBR1 and had intrauterine-onset destructive pancreatitis. In addition, we found that Ubr1−/− mice, whose previously reported phenotypes include reduced weight and behavioral abnormalities, had an exocrine pancreatic insufficiency, with impaired stimulus-secretion coupling and increased susceptibility to pancreatic injury. Our findings indicate that deficiency of UBR1 perturbs the pancreas' acinar cells and other organs, presumably owing to metabolic stabilization of specific substrates of the N-end rule pathway.
BackgroundRecruitment and migration of phagocytes to the site of inflammation are key events in the onset of inflammation. Albeit crucial for pathogen elimination, tissue repair and restoration of ...tissue homeostasis, dysregulated phagocyte infiltration can also cause severe inflammatory disorders. Therefore, targeting and modulation of phagocyte infiltration represents a promising new approach to fight inflammatory disorders and diseases, such as rheumatoid arthritis. Additionally, non-invasive tracking of phagocyte migration to the site of inflammation could extend both scientific knowledge as well as the repertoire of diagnostic strategies in clinical use.ObjectivesThe aim of this study was to establish a fluorescence reflectance imaging (FRI) based system to visualize and analyze migration properties of different cell populations in inflammatory disease models, like experimental arthritis, in vivo.MethodsImmortalized murine myeloid progenitor ER-HoxB8 cells were differentiated to neutrophils or monocytes (1). Cells were labeled with the membrane-selective fluorescent dyes DIR (2) or DID, respectively. We analyzed viability and functionality of stained cells in vitro and investigated their ability to migrate to sites of inflammation in vivo in several mouse models - particularly in a collagen induced arthritis (CIA) mouse model - via fluorescence reflectance imaging (FRI). Using CRISPR-Cas9 technology we introduced targeted gene deletions for main adhesion molecules.ResultsDifferentiated ER-HoxB8 cells could effectively be labeled with DIR or DID. Labeling of monocytes or neutrophils did not affect cellular viability or functionality in vitro. Subsequent in vivo imaging experiments allowed the visualization of migrated labeled phagocytes in different murine disease models, thereby cells could be detected at sites of inflammation with high sensitivity and specificity. In a CIA mouse model the amount of immigrated cells could even be associated closely to disease score and disease severity. Thus, the detection of immigration of labeled cells might also give hints about new inflammatory spots that are about to settle up before they can be detected macroscopically. Furthermore, differential cell labeling allowed direct quantitative comparison of differences in migration rates of wildtype and CD18 or CD49d knockout cells in vivo.ConclusionsSpecific and distinguishable labeling of diverse cell types allows in vivo tracking and subsequent quantification of migrated cells within the same animal. Targeted gene deletion allows analysis of molecular mechanisms relevant for leukocyte recruitment during different stages of arthritis. Correlation of the amount of immigrated cells to disease severity offers new opportunities to non-invasively detect and monitor inflammatory sites in vivo.References Wang GG, Calvo KR, Pasillas MP, Sykes DB, Häcker H, Kamps MP. Quantitative production of macrophages or neutrophils ex vivo using conditional Hoxb8. Nat Methods. 2006;3(4):287–93.Eisenblätter M, Ehrchen J, Varga G, Sunderkötter C, Heindel W, Roth J, et al. In vivo optical imaging of cellular inflammatory response in granuloma formation using fluorescence-labeled macrophages. J Nucl Med. 2009;50:1676–82. Disclosure of InterestNone declared
Aim of this study was to investigate the association of genetic variants of functional polymorphisms of matrix metalloproteinase and Cubilin (CUBN) with diabetic nephropathy (DN), end-stage renal ...disease (ESRD), and risk of cardiovascular disease (CVD) in Caucasian type 2 diabetes (T2D) patients.
472 T2D-patients were genotyped for 3 single-nucleotide polymorphisms (SNPs; MMP-2 rs2285053, MMP-9 rs17576 and CUBN rs1801239). Genotyping was carried out by allelic discrimination using TaqMan SNP-genotyping-assay.
MMP-9 (Gln279Arg) AA-genotype (OR 0.17 0.04-0.62, p = 0.008) and the time elapsed since diagnosis of T2D without onset of proteinuria (OR 0.87 0.79-0.97, p = 0.008) were found to be independently associated with reduced risk of susceptibility to DN. On the contrary higher stages of chronic kidney disease (OR 1.93 1.15-3.23, p = 0.012) and the presence of MMP-9 GG-genotype were independently associated with DN (OR 6.07 1.60-22.99, p = 0.008). The CUBN CC or C-risk-allele of rs1801239 was associated with ESRD (OR 2.04 1.07-3.87, p = 0.03) and peripheral artery disease (OR 2.08 1.12-3.88, p = 0.021). We could not find an association with MMP-2, MMP-9, or CUBN with CVD in a composite clinical endpoint model.
This study highlights that MMP-9 or CUBN-SNPs may exert effects on risk of susceptibility to DN or ESRD. We provide novel evidence on genetic susceptibility for macroangiopathy in patients with a missense variant of CUBN (Ile2984Val) in patients with T2D.
This paper reports on investigations of the beneficial effects of electron beam alloying (EBA) and electron beam dispersion alloying (EBDA) on the wear behavior of AZ91D Mg alloy under mild wear ...conditions with applied normal loads of 1…10 N. The layers generated had a thickness of 1.5 mm with Al contents of 30 wt.%. For dispersion alloyed layers, TiC was added with particle sizes of 20…100 µm. At a sliding distance of 20 m, the wear rates of alloyed layers (150 HB) and dispersion alloyed layers (180 HB) were almost the same and could be reduced by half compared to the untreated AZ91D (60 HB). Due to their large size and the large spaces between them, TiC particles were pressed into the layer matrix, or were torn out and acted as additional abrasives. Therefore, at a sliding distance of 50 m, the wear rate of dispersion alloyed layers increased to the level of the base material.
Sabine Zenker discusses best practice issues when using a 3-dimensional vector technique for rejuvenation of the mid-face
The facial ageing process is accompanied by a profound loss of volume, ...especially in the mid-face. Therefore, it is particularly important for a cosmetic surgeon to understand the changes to the mid-face 3-dimensionally, and then to treat with advanced injection techniques in such a way that earlier youthful facial proportions are restored, or existing proportions optimised. In this article, the so-called 'vector technique' will be discussed as it is particularly suitable for volume augmentation of the mid face. With the help of a volumising filler material, positive cranio-laterally directed vectors - acting against gravity - are placed in the fanning technique. The dermal filler based on calcium hydroxylapatite (Radiesse®) is particularly suitable for this approach as it allows a very distinct, precise lifting effect and - in contrast with hyaluronic acid based fillers - has no hygroscopic effect. In addition, the filling effect lasts for a long time in the majority of cases.
Abstract Introduction Noonan syndrome (NS) is characterized by dysmorphic facies, short stature and congenital heart defects. Various haemostatic disorders have been described in NS patients, but not ...all were related to bleeding, which itself is present in up to 65%. Several subgroups of NS - especially those with PTPN11 mutation - are associated with pulmonary stenosis. As it is known that some heart defects are prone to a shear stress related destruction of the von Willebrand factor as an important haemostatic component, we aimed to find out, whether the pulmonary stenosis could be responsible for such a mechanism in NS patients. Patients, Methods and Results We investigated the haemostatic system in 15 children with genetically proven NS (14 with PTPN11 , one with SOS1 mutation). Platelet count, basic coagulation parameters, fibrinogen and antithrombin were normal in all patients, none had a relevant reduction of coagulation factor activities. Five patients had pulmonary valve stenosis with systolic gradients > 60 mmHg. In three of them a deficiency of the high molecular weight multimers and a pathologic collagen-binding capacity were detected, suggesting acquired von Willebrand syndrome. Nine of our patients indicated a relevant bleeding diathesis and complained of easy bruising, three reported spontaneous gum bleeding. In conclusion the destruction of the von Willebrand factor could explain the bleeding in some of the NS patients with pulmonary valve stenosis. Our finding is of clinical relevance since most of these patients require either interventional cardiac catheterization or open heart surgery which may be complicated by the haemorrhagic tendency.
Numerous animal studies demonstrated the importance of components of the ephrin/Eph system for correct visual system development. Analogous investigations in humans are entirely missing. Here, we ...examined the visual system in humans with ephrin-B1 deficiency, which is x-linked and associated with the cranio-fronto-nasal syndrome (CFNS) in heterozygous females.
For one male hemizygous for ephrin-B1 deficiency and three affected heterozygous females with molecular-genetically confirmed mutations, the integrity of the partial decussation of the optic nerves was assessed with visual evoked potentials (VEPs) and compared with albinotic, achiasmic, and control participants with healthy vision. Further, retinal morphology and function and the gross-retinotopic representation of the primary visual cortex were examined with spectral-domain optical coherence tomography (SD-OCT), ERG, and multifocal (mf) VEPs for the male participant and part of the carriers.
Strabismus and lack of stereovision was evident in the male and two of the females. Other characteristics of the visual system organization and function were normal: (1) retina: SD-OCT and funduscopy indicated normal foveal and optic nerve head morphology. Electroretinograms indicated normal retinal function, (2) optic chiasm: conventional (c)VEP showed no evidence for misrouting and mfVEPs were only suggestive of, if any, very minor local misrouting, and (3) visual cortex: mfVEP characteristics indicated normal retinotopic gross-representations of the contralateral visual hemifield in each hemisphere.
While ephrin-B1 deficiency leads to abnormal visual pathways in mice, it leaves the human visual system, apart from deficits in binocular vision, largely normal. We presume that other components of the ephrin-system can substitute the lack of ephrin-B1 in humans.
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