Soluble Receptor for Advanced Glycation End Products (sRAGE) Is a Sensitive Biomarker in Human Pulmonary Arterial Hypertension.

Pulmonary arterial hypertension (PAH) is a progressive condition with an unmet need for early diagnosis, better monitoring, and risk stratification. The receptor for advanced glycation end products (RAGE) is activated in response to hypoxia and vascular injury, and is associated with inflammation, cell proliferation and migration in PAH. For the adult cohort, we recruited 120 patients with PAH, 83 with idiopathic PAH (IPAH) and 37 with connective tissue disease-associated PAH (CTD-PAH), and 48 controls, and determined potential plasma biomarkers by enzyme-linked immunoassay. The established heart failure marker NTproBNP and IL-6 plasma levels were several-fold higher in both adult IPAH and CTD-PAH patients versus controls. Plasma soluble RAGE (sRAGE) was elevated in IPAH patients (3044 ± 215.2 pg/mL) and was even higher in CTD-PAH patients (3332 ± 321.6 pg/mL) versus controls (1766 ± 121.9 pg/mL; p < 0.01). All three markers were increased in WHO functional class II+III PAH versus controls (p < 0.001). Receiver-operating characteristic analysis revealed that sRAGE has diagnostic accuracy comparable to prognostic NTproBNP, and even outperforms NTproBNP in the distinction of PAH FC I from controls. Lung tissue RAGE expression was increased in IPAH versus controls (mRNA) and was located predominantly in the PA intima, media, and inflammatory cells in the perivascular space (immunohistochemistry). In the pediatric cohort, plasma sRAGE concentrations were higher than in adults, but were similar in PH (n = 10) and non-PH controls (n = 10). Taken together, in the largest adult sRAGE PAH study to date, we identify plasma sRAGE as a sensitive and accurate PAH biomarker with better performance than NTproBNP in the distinction of mild PAH from controls

Severe infections of Panton-Valentine leukocidin positive Staphylococcus aureus in children

Infections caused by Panton-Valentine leukocidin-positive Staphylococcus aureus (PVL-SA) mostly present as recurrent skin abscesses and furunculosis. However, life-threatening infections (eg, necrotizing pneumonia, necrotizing fasciitis, and osteomyelitis) caused by PVL-SA have also been reported.We assessed the clinical phenotype, frequency, clinical implications (surgery, length of treatment in hospitals/intensive care units, and antibiotic treatments), and potential preventability of severe PVL-SA infections in children.Total, 75 children treated for PVL-SA infections in our in- and outpatient units from 2012 to 2017 were included in this retrospective study.Ten out of 75 children contracted severe infections (PVL-methicillin resistant S aureus n = 4) including necrotizing pneumonia (n = 4), necrotizing fasciitis (n = 2), pyomyositis (n = 2; including 1 patient who also had pneumonia), mastoiditis with cerebellitis (n = 1), preorbital cellulitis (n = 1), and recurrent deep furunculosis in an immunosuppressed patient (n = 1). Specific complications of PVL-SA infections were venous thrombosis (n = 2), sepsis (n = 5), respiratory failure (n = 5), and acute respiratory distress syndrome (n = 3). The median duration of hospital stay was 14 days (range 5-52 days). In 6 out of 10 patients a history suggestive for PVL-SA colonization in the patient or close family members before hospital admission was identified.PVL-SA causes severe to life-threatening infections requiring lengthy treatments in hospital in a substantial percentage of symptomatic PVL-SA colonized children. More than 50% of severe infections might be prevented by prompt testing for PVL-SA in individuals with a history of abscesses or furunculosis, followed by decolonization measures

Functional characterization of hypomorphic protein variants in Nijmegen Breakage Syndrome

Das Nijmegen Breakage Syndrom gehört zur Gruppe der DNA-Reparaturdefekte und ist klinisch durch das gehäufte Auftreten von Tumoren im jungen Lebensalter, eine zelluläre und humorale Immunschwäche sowie eine Mikrozephalie gekennzeichnet. In mehr als 90% der Fälle wird das Syndrom durch die Mutation 657del5 im NBN-Gen im homozygoten Zustand verursacht, die zu einem hypomorphen carboxyterminalen Protein (p70-Nibrin) führt. Dieses hat eine residuale Funktion, die für das zelluläre Überleben essentiell ist. NBN wird als haploinsuffizientes Tumorsuppressorgen interpretiert, da auch bei hetrozygoten Merkmalsträgern ein erhöhtes Auftreten maligner Erkrankungen beobachtet wird. Eine Reihe von NBN-Missense-Mutationen sind bei verschiedenen malignen Erkrankungen beschrieben. Diese sind innerhalb oder in unmittelbarer Nähe von funktionell wichtigen Domänen des Nibrins lokalisiert. Die in Zusammenhang mit einem besonders milden Phänotyp beschriebene 742-743insGG-Mutation resultiert in Folge von alternativem Splicing in einem Protein mit einer internen Deletion und intaktem Amino- und Carboxyterminus (p80-Nibrin). In dieser Arbeit wurde anhand von Interaktionsstudien die Proteinfunktion der unterschiedlichen Varianten charakterisiert und verglichen. Durch Transfektion mit Antisense-Oligunkleotiden lässt sich das p80-Nibrin in NBS-Patientenzellen anreichern, was in dieser Arbeit regelhaft angewendet wurde. Wir konnten zeigen, dass das p70- und auch p80-Nibrin nicht mit bekannten aminoterminalen Interaktionspartnern in Wechselwirkung treten (MDC1, γH2AX oder CtIP). Daraus leitet sich ab, dass ein intakter Aminoterminus für diese Proteininteraktionen notwendig ist. Die Experimente mit Nibrinvarainten, die aus aminoterminal lokalisierten Missensemutationen resultieren, belegen diese Annahme. Diese Ergebnisse legen nahe, dass das durch Aminosäueraustausche verminderte Interaktionspotential zur Tumorassoziation führt. Experimente zur Analyse von Proteinwechselwirkungen mit quantitativer Massenspektrometrie nach Affinitätsaufreinigung lieferten Hinweise dafür, dass die carboxyterminal vermittelte Wechselwirkung mit Bestandteilen des MRN-Komplexes (Mre11, Rad50) mit p80- (und auch mit wt-/p95-Nibrin) im Vergleich zum p70-Nibrin deutlich stabilisiert ist. Über die Interaktion mit Mre11 und Rad50 nimmt Nibrin eine zentrale Rolle bei der Reparatur von DNA-Doppelstrangbrüchen ein. Weiterhin wurden potentielle Interaktionspartner identifiziert, die das p80- und p95-Nibrin gengenüber dem p70-Nibrin favorisieren. Einige dieser Proteine können funktionell mit der DNA-Reparatur oder der Zellzyklusregulation in Verbindung gebracht werden. Diese Ergebnisse zeigen, dass die funktionelle Überlegenheit des p80-Nibrins in einer Stabilisation des MRN-Komplexes liegen kann. Außerdem wurden Proteine identifiziert, welche potentielle Interaktionspartner des p80-Nibrins sind und daher den assoziierten milderen Phänotyp begründen könnten. Diese Daten liefern pathomechanistische Erklärungsansätze für die variierenden Phänotypen bei unterschiedlichen NBN- Mutationen und legen nahe, dass die hier verwendete Behandlung mit Anitisense- Oligonukleotiden als innovativer therapeutischer Ansatz beim NBS genutzt werden könnte.Nijmegen breakage syndrome is a DNA repair deficiency disorder whose clinical features include increased cancer susceptibility, cellular and humoral immunodeficiency and microcephaly. The vast majority of cases are caused by the homozygous NBN mutation 657del5, which results in a hypomorphic protein representing the carboxy-terminal two-thirds of the protein (p70-nibrin). This protein has been found to be essential for cellular survival due to its residual function. An increased incidence of malignant diseases has also been observed in heterozygous individuals. Therefore, NBN is considered to be a haploinsufficient tumorsupressor gene. Various NBN missense mutations have been reported to be associated with a number of different malignancies. The resulting altered amino acids are located within or adjacent to important functional protein domains. A distinctively mild NBS-phenotype is associated with the 742-743insGG mutation which results via alternative splicing in a protein variant containing an internal deletion but including both the amino and carboxy termini (p80-nibrin). In the experiments described here, the protein function of the different nibrin variants was characterized and compared using protein interaction studies. In NBS-cells, p80-nibrin can be generated after transfection of specific antinsense oligonucleotides. We were able to show that neither p70- nor p80-nibrin interact with previously described amino-terminal binding partners (MDC1, γH2AX, CtIP). This illustrates the importance of an intact amino-terminus for these interactions. The experiments with nibrin variants containing missense mutations substantiate this assumption and suggest that loss of protein-protein interactions may account for the association of these mutations with tumour occurrence. The formation of the MRN-complex (mre11, rad50, nibrin) plays an essential role in DNA-double-strand-break repair. Affinity purification followed by mass spectrometry (q-AP-MS) studies indicate that the MRN-complex containing p80- (and p95-) nibrin is more stable than the complex containing p70-nibrin. Moreover, various proteins interacting preferentially with p80- and p95-nibrin could be identified. Some of these proteins are functionally related to DNA repair and cell cycle control. The results of this study show that the stabilization of the MRN-complex can lead to a functional advantage for p80-nibrin. Furthermore the identified putative interaction partners are possibly related to the milder phenotype of the patient with the 742-743insGG mutation. The data presented provide a molecular model for phenotypic variation related to different NBN-mutations and demonstrate the potential implementation of antisense oligonucleotides as an innovative therapeutic strategy in NBS

Slice2Volume: Fusion of multimodal medical imaging and light microscopy data of irradiation-injured brain tissue in 3D.

The dataset contains comprehensive image data for a total of nine mice, which underwent normal tissue brain irradiation with 90 MeV protons. In particular, the image data comprise cone-bem computed tomographies (CBCT), Monte Carlo beam transport simulations based on those CTs, regular magnetic resonance imaging (MRI) follow-up (≥ 26 weeks), a co-aligned DSURQE mouse brain atlas and scanned whole-brain tissue sections with histochemical and immunofluorescent markers for morphology (H&E), cell nuclei (DAPI), astrocytes (GFAP), microglia (Iba1), the intermediate filament protein Nestin, proliferation (Ki67), neurons (NeuN) and oligodendrocytes (OSP). The volumetric image data (i.e. CBCT, MRI and brain atlas) were co-aligned using the ImageJ plugin Big Warp. The CBCT data was used as spatial reference to allow for mask-based, slice-wise alignment of CBCT and light microscopy image data in 3D with the scriptable registration tool Elastix. We provide the data in raw format and as aligned data sets, as well as their spatial transformations

Slice2Volume: Fusion of multimodal medical imaging and light microscopy data of irradiation-injured brain tissue in 3D.

The dataset contains comprehensive image data for a total of nine mice, which underwent normal tissue brain irradiation with 90 MeV protons. In particular, the image data comprise cone-bem computed tomographies (CBCT), Monte Carlo beam transport simulations based on those CTs, regular magnetic resonance imaging (MRI) follow-up (≥ 26 weeks), a co-aligned DSURQE mouse brain atlas and scanned whole-brain tissue sections with histochemical and immunofluorescent markers for morphology (H&E), cell nuclei (DAPI), astrocytes (GFAP), microglia (Iba1), the intermediate filament protein Nestin, proliferation (Ki67), neurons (NeuN) and oligodendrocytes (OSP). The volumetric image data (i.e. CBCT, MRI and brain atlas) were co-aligned using the ImageJ plugin Big Warp. The CBCT data was used as spatial reference to allow for mask-based, slice-wise alignment of CBCT and light microscopy image data in 3D with the scriptable registration tool Elastix. We provide the data in raw format and as aligned data sets, as well as their spatial transformations.Chunked zip: The histological data are stored as chunked .zip files (*.zip.001 - *.zip.0XX). In order to unpack the data, download all chunks into the same directory, then unpack

Exome sequencing helped the fine diagnosis of two siblings afflicted with atypical Timothy syndrome (TS2)

BACKGROUND: Long-QT syndrome (LQTS) causes a prolongation of the QT-interval in the ECG leading to life threatening tachyarrhythmia and ventricular fibrillation. One atypical form of LQTS, Timothy syndrome (TS), is associated with syndactyly, immune deficiency, cognitive and neurological abnormalities as well as distinct cranio-facial abnormalities. CASE PRESENTATION: On a family with both children diagnosed with clinical LQTS, we performed whole exome sequencing to comprehensively screen for causative mutations after a targeted candidate gene panel screen for Long-QT syndrome target genes failed to identify any underlying genetic defect. Using exome sequencing, we identified in both affected children, a p.402G > S mutation in exon 8 of the CACNA1C gene, a voltage-dependent Ca(2+) channel. The mutation was inherited from their father, a mosaic mutation carrier. Based on this molecular finding and further more careful clinical examination, we refined the diagnosis to be Timothy syndrome (TS2) and thereby were able to present new therapeutic approaches. CONCLUSIONS: Our study highlights the difficulties in accurate diagnosis of patients with rare diseases, especially those with atypical clinical manifestation. Such challenge could be addressed with the help of comprehensive and unbiased mutation screening, such as exome sequencing