Increased Intracranial Arterial Pulsatility and Microvascular Brain Damage in Pseudoxanthoma Elasticum.

BACKGROUND AND PURPOSE: Carotid siphon calcification might contribute to the high prevalence of cerebrovascular disease in pseudoxanthoma elasticum through increased arterial flow pulsatility. This study aimed to compare intracranial artery flow pulsatility, brain volumes, and small-vessel disease markers between patients with pseudoxanthoma elasticum and controls and the association between arterial calcification and pulsatility in pseudoxanthoma elasticum. MATERIALS AND METHODS: Fifty patients with pseudoxanthoma elasticum and 40 age- and sex-matched controls underwent 3T MR imaging, including 2D phase-contrast acquisitions for flow pulsatility in the assessment of ICA and MCA and FLAIR acquisitions for brain volumes, white matter lesions, and infarctions. All patients with pseudoxanthoma elasticum underwent CT scanning to measure siphon calcification. Flow pulsatility (2D phase-contrast), brain volumes, white matter lesions, and infarctions (3D T1 and 3D T2 FLAIR) were compared between patients and controls. The association between siphon calcification and pulsatility in pseudoxanthoma elasticum was tested with linear regression models. RESULTS: Patients with pseudoxanthoma elasticum (mean age, 57 [SD, 12] years; 24 men) had significantly higher pulsatility indexes (1.05; range, 0.94-1.21 versus 0.94; range, 0.82-1.04; P = .02), lower mean GM volumes (597 [SD, 53] mL versus 632 [SD, 53] mL; P < .01), more white matter lesions (2.6; range, 0.5-7.5 versus 1.1; range, 0.5-2.4) mL; P = .05), and more lacunar infarctions (64 versus 8, P = .04) than controls (mean age, 58 [SD, 11] years; 20 men). Carotid siphon calcification was associated with higher pulsatility indexes in patients with pseudoxanthoma elasticum (ß = 0.10; 95% CI, 0.01-0.18). CONCLUSIONS: Patients with pseudoxanthoma elasticum have increased intracranial artery flow pulsatility and measures of small-vessel disease. Carotid siphon calcification might underlie the high prevalence of cerebrovascular disease in pseudoxanthoma elasticum.

Efficacy of DiaLife, an education program for relatives of adult patients with diabetes - study protocol of a cluster randomized controlled trial.

BACKGROUND: The global prevalence of diabetes mellitus (DM) has been increasing over recent decades. In Germany, the prevalence for DM type 1 and type 2 in adults is estimated at about 7.7%. Hence, diabetes has to be classified as a serious public health concern. Being diagnosed with DM and facing possible sequelae might have a negative impact on patients' mental and physical well-being. However, diabetes not only affects patients themselves, but also their close relatives. To improve the quality of life for patients and relatives alike, the German Association of Diabetes Nurses and Education experts (VDBD) elaborated the first education program tailor-made for relatives of diabetes patients. This article describes the concept and design of the trial evaluating the efficacy of this education program called "DiaLife-Living Together with Diabetes". METHODS: This evaluation study is a cluster randomized controlled trial, in which the study centers will be randomly assigned either to the intervention group or the control group. Study centers will recruit relatives of and patients with DM type 1 and type 2. Members of the intervention group will participate in the education program DiaLife, whereas participants randomized in the control group will act as waiting-list controls. The study will assess the efficacy of DiaLife by comparing diabetes-related knowledge between the intervention and control groups as the primary outcome for participants. As the primary outcome in patients, the Hba1c value will be assessed. In addition, diabetes-related distress, family interaction, and other secondary endpoints will be considered as secondary outcomes. Long-term efficacy will be assessed 6 and 12 months after intervention. Hierarchical regression models will be used to analyze effects over time. DISCUSSION: While there is scientific evidence for the efficacy of education programs addressed to (diabetes) patients, there is a research gap with regard to intervention studies evaluating the efficacy of education programs designed for patients' relatives. The study results will provide information on the efficacy of the DiaLife education program. In addition, factors that might hinder a successful implementation of an education program for relatives will be identified. TRIAL REGISTRATION: German Clinical Trials Register, DRKS00015157 . Registered on 24 August 2018.

The Effects of Iodine Attenuation on Pulmonary Nodule Volumetry using Novel Dual-Layer Computed Tomography Reconstructions.

OBJECTIVES: To assess the effect of iodine attenuation on pulmonary nodule volumetry using virtual non-contrast (VNC) and mono-energetic reconstructions. METHODS: A consecutive series of patients who underwent a contrast-enhanced chest CT scan were included. Images were acquired on a novel dual-layer spectral CT system. Conventional reconstructions as well as VNC and mono-energetic images at different keV levels were used for nodule volumetry. RESULTS: Twenty-four patients with a total of 63 nodules were included. Conventional reconstructions showed a median (interquartile range) volume and diameter of 174 (87 - 253) mm3 and 6.9 (5.4 - 9.9) mm, respectively. VNC reconstructions resulted in a significant volume reduction of 5.5% (2.6 - 11.2%; p<0.001). Mono-energetic reconstructions showed a correlation between nodule attenuation and nodule volume (Spearman correlation 0.77, (0.49 - 0.94)). Lowering the keV resulted in increased volumes while higher keV levels resulted in decreased pulmonary nodule volumes compared to conventional CT. CONCLUSIONS: Novel dual-layer spectral CT offers the possibility to reconstruct VNC and mono-energetic images. Those reconstructions show that higher pulmonary nodule attenuation results in larger nodule volumes. This may explain the reported underestimation in nodule volume on non-contrast enhanced compared to contrast-enhanced acquisitions. KEY POINTS: • Pulmonary nodule volumes were measured on virtual non-contrast and mono-energetic reconstructions • Mono-energetic reconstructions showed that higher attenuation results in larger volumes • This may explain the reported nodule volume underestimation on non-contrast enhanced CT • Mostly metastatic pulmonary nodules were evaluated, results might differ for benign nodules.

Can routine chest radiography be used to diagnose mild COPD? A nested case-control study.

OBJECTIVES: To determine whether mild stage chronic obstructive pulmonary disease (COPD) can be detected on chest radiography without substantial overdiagnosis. METHODS: A retrospective nested case-control study (case:control, 1:1) was performed in 783 patients scheduled for cardiothoracic surgery who underwent both spirometry and a chest radiograph preoperative. Diagnostic accuracy of chest radiography for diagnosing mild COPD was investigated using objective measurements and overall appearance specific for COPD on chest radiography. Inter-observer variability was investigated and variables with a kappa >0.40 as well as baseline characteristics were used to make a diagnostic model which was aimed at achieving a high positive predictive value (PPV). RESULTS: Twenty percent (155/783) had COPD. The PPV of overall appearance specific for COPD alone was low (37-55%). Factors in the diagnostic model were age, type of surgery, gender, distance of the right diaphragm apex to the first rib, retrosternal space, sternodiaphragmatic angle, maximum height right diaphragm (lateral view) and subjective impression of COPD (using both views). The model resulted in a PPV of 100%, negative predictive value (NPV) of 82%, sensitivity of 10% and specificity of 100% with an area under the curve of 0.811. CONCLUSIONS: Detection of mild COPD without substantial overdiagnosis was not feasible on chest radiographs in our cohort.

Imaging of pediatric great vessel stents: Computed tomography or magnetic resonance imaging?

BACKGROUND: Complications might occur after great vessel stent implantation in children. Therefore follow-up using imaging is warranted. PURPOSE: To determine the optimal imaging modality for the assessment of stents used to treat great vessel obstructions in children. MATERIAL AND METHODS: Five different large vessel stents were evaluated in an in-vitro setting. All stents were expanded to the maximal vendor recommended diameter (20mm; n = 4 or 10mm; n = 1), placed in an anthropomorphic chest phantom and imaged with a 256-slice CT-scanner. MRI images were acquired at 1.5T using a multi-slice T2-weighted turbo spin echo, an RF-spoiled three-dimensional T1-weighted Fast Field Echo and a balanced turbo field echo 3D sequence. Two blinded observers assessed stent lumen visibility (measured diameter/true diameter *100%) in the center and at the outlets of the stent. Reproducibility of diameter measurements was evaluated using the intraclass correlation coefficient for reliability and 95% limits of agreement for agreement analysis. RESULTS: Median stent lumen visibility was 88 (IQR 86-90)% with CT for all stents at both the center and outlets. With MRI, the T2-weighted turbo spin echo sequence was preferred which resulted in 82 (78-84%) stent lumen visibility. Interobserver reliability and agreement was good for both CT (ICC 0.997, mean difference -0.51 [-1.07-0.05] mm) and MRI measurements (ICC 0.951, mean difference -0.05 [-2.52 --2.41] mm). CONCLUSION: Good in-stent lumen visibility was achievable in this in-vitro study with both CT and MRI in different great vessel stents. Overall reliability was good with clinical acceptable limits of agreement for both CT and MRI. However, common conditions such as in-stent stenosis and associated aneurysms were not tested in this in-vitro study, limiting the value of the in-vitro study.

[Pathogenesis and molecular pathology of vestibular schwannoma]. / Pathogenese und Molekularpathologie des Vestibularisschwannoms.

Schwannomas are benign Schwann cell-derived tumors of the peripheral nerve sheath often involving the vestibular cranial nerve (vestibular schwannoma). Histologically, they consist of bipolar spindle cells and show a moderate cellularity. Typically, Antoni A regions with a storiform pattern and loose Antoni B regions are intermingled. Verocay bodies are the pathognomonic palisading structures. Malignant transformation is rare. Merlin (schwannomin), the protein product of NF2, is inactivated by mutations, loss of heterozygosity or methylation. Within neurofibromatosis type 2, a germline mutation is present in about half of cases, whereas tumors demonstrate an additional second hit of the NF2 gene. A loss of chromosome 22 or 22q is common. Merlin links the cell membrane with the cytoskeleton and regulates intracellular signaling pathways leading to dysorganization when merlin is inactivated. Loss of merlin activates Rac1 and Ras, and the PAK1, mTORC1, EGFR-Ras-ERK, PI3K-Akt, WNT and Hippo pathways as well as receptor tyrosine kinases. Furthermore, merlin locates to the nucleus and inhibits E3 ubiquitin ligase CRL4DCAF1. Besides biallelic inactivation of NF2 in schwannomas, other genes are involved in the pathogenesis of schwannomatosis-associated schwannomas such as LZTR1, SMARCB1, COQ6 indicating an important role of SWI/SNF chromatin-remodeling complex for schwannoma development. Our own investigations point to deregulation of BAF170, another essential SWI/SNF complex component. Knowledge of mechanisms allows targeted molecular therapy, especially in vestibular schwannomas, using antagonists against mTOR (rapamycin/sirolmus/everolimus), EGFR (lapatinib) or VEGF (bevacizumab), although clinical studies have been in part disappointing so far.

The Biochemistry of Haemonchus contortus and Other Parasitic Nematodes.

Different life cycle stages of Haemonchus contortus adapt to different ecosystems. This adaptation is accompanied by alterations in gene transcription and expression associated with the energy, amino acid, nitrogen, lipid and/or nucleic acid metabolism of the respective stages. For example, the aerobic metabolism of larvae depends on an efficient citric acid cycle, whereas the anaerobic metabolism of adults requires glycolysis, resulting in the production of volatile fatty acids, such as acetic acid and propionic acid. There are only few anthelmintics targeting nematode energy metabolism. In addition, H. contortus has reduced pathways for amino acid metabolism, polyamine metabolism and nitrogen excretion pathways. Moreover, nucleic acid metabolism comprising purine and pyrimidine salvage pathways as well as lipid metabolism are reduced. In addition, nematodes possess a particular composition of their cuticle. Energy production of adult worms is mainly linked to egg production and complex regulation of the neuromuscular system in both females and males. In this context, microtubules consisting of α- and ß-tubulin heterodimers play a crucial role in the presynaptic vesicle transport. Due to the significant distinction of its quarternary structure in nematodes in comparison to other organisms, ß-tubulin was identified as a major target for benzimidazoles used for anthelmintic treatment. Concerning the function of the neuromuscular system, acetylcholine, a ligand of the nicotinic acetylcholine receptor (nAChR), is the major excitatory neurotransmitter in H. contortus. In contrast, glutamate-gated chloride channels, calcium- and voltage-dependent potassium channels as well as γ-aminobutyric acid (GABA)A and its receptors act as inhibitory neurotransmitters and thus opponents to nAChR. For example, the calcium- and voltage-dependent potassium channel SLO-1 is an important target of emodepside, which is involved in the sensitive regulation of activatory and inhibitory receptors of the nervous system. Most of the modern anthelmintics target these different neuromuscular receptors. The mechanisms of resistance to anthelmintics, either specific or non-specific, are associated with changes in the molecular targets of the drugs, changes in metabolism of the drug (inactivation, removal or prevention of its activation) and/or increased efflux systems. The biochemical and molecular analyses of key developmental, metabolic and structural process of H. contortus still require substantial efforts. The nAChR, glutamate-gated chloride channel and calcium- and voltage-dependent potassium channel SLO-1 have long been known as being essential for nematode survival. Therefore, future research should be intensified to fully resolve the three-dimensional structures of these receptors, as has already been started for glutamate-gated chloride channel. With this knowledge, it should be possible to design new anthelmintics, which possess improved binding capacities to corresponding receptors.

New horizons in cardiac CT.

Until recently, cardiovascular computed tomography angiography (CCTA) was associated with considerable radiation doses. The introduction of tube current modulation and automatic tube potential selection as well as high-pitch prospective ECG-triggering and iterative reconstruction offer the ability to decrease dose with approximately one order of magnitude, often to sub-millisievert dose levels. In parallel, advancements in computational technology have enabled the measurement of fractional flow reserve (FFR) from CCTA data (FFRCT). This technique shows potential to replace invasively measured FFR to select patients in need of coronary intervention. Furthermore, developments in scanner hardware have led to the introduction of dual-energy and photon-counting CT, which offer the possibility of material decomposition imaging. Dual-energy CT reduces beam hardening, which enables CCTA in patients with a high calcium burden and more robust myocardial CT perfusion imaging. Future-generation CT systems will be capable of counting individual X-ray photons. Photon-counting CT is promising and may result in a substantial further radiation dose reduction, vastly increased spatial resolution, and the introduction of a whole new class of contrast agents.

Infant with unusually large choroid plexus papilloma undergoing emergency surgery. Case report with special emphasis on the surgical strategy.

Choroid plexus papillomas are one of the most common tumors of the nervous system in infants. The most frequent early symptoms, megalocephalia and vomiting, caused by elevated intracranial pressure often lead to a diagnosis only at a critical clinical stage. This study describes a case of a 3-month-old infant with a choroid plexus papilloma measuring 7 x 8 x 6 cm originating in the right lateral ventricle. The infant underwent emergency surgery in an acutely deteriorated state, i.e., acute herniation symptoms with fixed and dilated pupils. Despite of the size of the tumor, the proximity to eloquent cortex, and clinically deteriorated state, the infant recovered completely.

Protein profiling of bladder cancer using the 2D-PAGE and SELDI-TOF-MS technique.

Protein profiling is a promising tool for tumor characterization and the detection of tumor markers in bladder cancer. Techniques for 2-dimensional polyacrylamide gel electrophoresis (2D-PAGE) and surface-enhanced laser desorption/ionization with time-of-flight mass spectrometry (SELDI-TOF-MS) have improved; both were evaluated using bladder tumor tissue. Normal urothelium and pTa G2, pT1 G3, and >or=pT3 G3 tissues were obtained from the operating room and, after macrodissection, subjected to 2D-PAGE and to SELDI-TOF-MS ProteinChip. 2D-PAGE gels expressed significantly different protein patterns for pTa G2 and pT3 G3 tumors. pT1 G3 tumors showed expression profiles similar to those of the invasive tumors, with upregulation of galectin 3, gelsolin, villin 2, moesin, and annexin 6. Similarly, distinct protein peaks were detected for superficial and muscle-invasive urothelial cancers by SELDI-TOF-MS. Six of seven superficial pTa G2 tumors showed an intense peak at 6.7 and 10.1 kD, while invasive carcinomas showed an intense peak near 9.5 kD. No disturbing influence of surrounding tissue on the results was detected. It was shown that both techniques (2D-PAGE and ProteinChip) work well, and especially ProteinChip analysis seems promising for clinical application.

Mechanisms of action of emodepside.

The research of the class of cyclic octadepsipeptides started at the beginning of the 1990s. PF1022A, the starting material of emodepside, is a natural secondary metabolite of the fungus Mycelia sterilia, which belongs to the microflora of the leaves of Camellia japonica. PF1022A consists of four N-methyl-L-leucins, two D-Iactic acids and two D-phenyllactic acids, which build up a cyclic octadepsipeptide with an alternating L-D-L-configuration. Emodepside is a semisynthetic derivative of PF1022A, which contains a morpholine attached in para position at each of both D-phenyllactic acids. Emodepside is efficacious against a variety of gastrointestinal nematodes. Emodepside binds to a presynaptic latrophilin receptor in nematodes. The following presynaptic signal transduction occurs via activation of Gqalpha protein and phospholipase-Cbeta, which leads to mobilization of diacylglycerol (DAG). DAG then activates UNC-13 and synaptobrevin, two proteins which play an important role in presynaptic vesicle-functioning. This finally leads to the release of a currently unidentified transmitter. The transmitter (or modulator) exerts its effects at the postsynaptic membrane and induces a flaccid paralysis of the pharynx and the somatic musculature in nematodes.

Influence of the cyclooctadepsipeptides PF1022A and PF1022E as natural products on the design of semi-synthetic anthelmintics such as emodepside.

The 24-membered cyclooctadepsipeptide (CODP) PF1022A, the active metabolite of the fungus imperfectus Mycelia sterilia (Rosellinia sp.) isolated from the plant Camellia japonica in Japan, is described as a powerful broad-spectrum anthelmintic natural product with low toxicity in animals. Further CODPs such as PF1022B, C, D and E have been isolated from the same culture and their structures have been established. Both PF1022A and PF1022E serve as valuable starting materials for the synthesis of semi-synthetic CODP derivatives with improved intrinsic anthelmintic potency and broad-spectrum activity. It was found that in most cases the di-substituted PF1022A derivatives showed a greater (or equal) activity by oral application against the gastrointestinal nematode Haemonchus contortus compared to the corresponding mono-substituted PF1022A analogues as exemplified by emodepside. In order to get additional information on the bioactive conformation, emodepside was transformed into its mono- and tetra-thionated derivatives by isosteric replacement. In the light of the increased efficacy of these derivatives against H. contortus or Trichostrongylus colubriformis, it has been suggested that the asymmetric conformation clearly influences the anthelmintic activity of CODPs. Although useful synthetic pathways are available today for the preparation of the semi-synthetic CODP emodepside, the fermentative production of its bis-para-nitro and bis-para-amino precursors could be the process used for its industrial-scale production in the future.

Cyclohexadepsipeptides (CHDPs) with improved anthelmintical efficacy against the gastrointestinal nematode (Haemonchus contortus) in sheep.

Besides 24-membered cyclooctadepsipeptides (CODPs) with the most prominent member of this class emodepside, the structurally related 18-membered cyclohexadepsipeptides (CHDPs) were of interest with regard to their efficacy against the nematode H. contortus in sheep.The CHDPs prepared by a simple total synthesis represent enniatin derivatives with strong in vivo activity against H. contortus in sheep. The correlation between the nature of the CHDP major conformers and their anthelmintic activities was studied in detail. All CHDPs with strong in vivo activity exists in deuterochloroform solution as conformers with restricted flexibility which was found by 2D-NMR spectroscopic analysis. This reduced flexibility of the major conformer can be exemplified by CHDPs containing e.g.: (i) an unsymmetrically folded conformation with no cis-amide bound, (ii) an internal hydrogen bond or (iii) one cis-amide bond, respectively.The strong in vivo anthelmintic activity against H. contortus in sheep indicates that the stereochemistry in 2-position of CHDPs is less important for their high inding affinity. It may be assumed that the identified inflexible region of the major conformers might mimic the active conformation of these CHDPs, which could be helpful for rational design of anthelmintics with less complicated structures.

Putative G protein-coupled receptors in parasitic nematodes--potential targets for the new anthelmintic class cyclooctadepsipeptides?

Parasitic nematodes cause major problems in livestock animals. Resistances to the most commonly used drugs are arising. The cyclooctadepsipeptide emodepside belongs to a new class of anthelmintics. A receptor for emodepside, Hc110-R, was previously identified in Haemonchus contortus. We have identified the complete coding sequences of putative orthologues in Cooperia oncophora and Ostertagia ostertagi, tri-chostrongyles in cattle. The putative receptors were named depsiphilins. The deduced amino acid sequence of C. oncophora depsiphilin has a similarity of 91% to the O. ostertagi sequence. The similarity of both the C. oncophora and O. ostertagi depsiphilin to Hc110-R is 89%, based on the amino acid sequence. The depsiphilins share 46% identity with the latrophilin-like protein 1 in Caenorhabditis elegans and 47% identity with a hypothetical protein in Caenorhabditis briggsae. Hc110-R and the latrophilin-like proteins of C. elegans were previously reported to be putative G protein-coupled receptors (GPCRs) and to be related to mammalian latrophilins. A seven transmembrane domain, a GPCR proteolytic site, and other conserved domains characteristic of receptors of the latrophilin group were identified within the depsiphilins. Therefore it seems reasonable to allocate the depsiphilins to the previously described latrophilins and latrophilin-like proteins.

Effects of a combinations of emodepside and praziquantel on parasites of reptiles and rodents.

A new combination of two anthelmintic compounds containing emodepside and praziquantel (Profender, Bayer AG, Levekusen, Germany) was tested in pet rodents and reptiles. Topical application of the two compounds led to the quick disappearance of nematodes and cestodes from a broad spectrum of hosts including mice, jirds, snakes, anole lizards, turtles, monitor lizards, etc. In reptiles the dosage had to be increased, since the thick outer layer of the epidermis hinders the penetration of the compounds. In animals with an extremely thick epidermis (e.g. monitor lizards, leguans) the new product was applied under the armpits.

Identification of Dll1 (Delta1) target genes during mouse embryogenesis using differential expression profiling.

The Notch signaling pathway has pleiotropic functions during mammalian embryogenesis. It is required for the patterning and differentiation of the presomitic and somitic paraxial mesoderm and of the neural tube. We used DNA-chip expression profiling and 2D-gel electrophoresis combined with peptide mass fingerprinting to identify genes and proteins differentially regulated in E10.5 Dll1 (delta-like 1, Delta1) mutant embryos. The differential expression profiling approach identified 47 regulated transcripts and 40 differentially expressed proteins. The majority of these genes has until now not been associated with Notch signaling. Subsequent whole-mount in situ hybridization confirmed that a subset of the identified transcripts has restricted and distinct patterns of expression in E10.5 mouse embryos. For most genes these expression patterns were affected in the presomitic mesoderm, in differentiating somites of Dll1 mutant embryos and in the neural tube and cells differentiating from it. Similar effects were observed in embryos homozygous for the Headturner (Htu) and pudgy (pu) mutations, which are alleles of the Notch ligands Jag1 and Dll3. The regulated expression of a subset of the proteins was validated by immunoblots. Remarkably six of the proteins down-regulated in Dll1 mutant embryos are proteasome subunits. The large set of regulated genes identified in this differential expression profiling approach is an important resource for further functional studies.

Identification of coexpressed gene clusters in a comparative analysis of transcriptome and proteome in mouse tissues.

A major advantage of the mouse model lies in the increasing information on its genome, transcriptome, and proteome, as well as in the availability of a fast growing number of targeted and induced mutant alleles. However, data from comparative transcriptome and proteome analyses in this model organism are very limited. We use DNA chip-based RNA expression profiling and 2D gel electrophoresis, combined with peptide mass fingerprinting of liver and kidney, to explore the feasibility of such comprehensive gene expression analyses. Although protein analyses mostly identify known metabolic enzymes and structural proteins, transcriptome analyses reveal the differential expression of functionally diverse and not yet described genes. The comparative analysis suggests correlation between transcriptional and translational expression for the majority of genes. Significant exceptions from this correlation confirm the complementarities of both approaches. Based on RNA expression data from the 200 most differentially expressed genes, we identify chromosomal colocalization of known, as well as not yet described, gene clusters. The determination of 29 such clusters may suggest that coexpression of colocalizing genes is probably rather common.

Efficacy of two cyclooctadepsipeptides, PF1022A and emodepside, against anthelmintic-resistant nematodes in sheep and cattle.

Resistance against the major currently available anthelmintics has reached a critical level in many small ruminant herds world-wide, and is increasingly becoming a problem in horses and cattle. Therefore, new products with different modes of action are urgently needed. Recently, such a new class of compounds, the anthelmintically active cyclooctadepsipeptides, was described. Here, the effects of cyclooctadepsipeptides on benzimidazole-, levamisole- and ivermectin-resistant populations of Haemonchus contortus in sheep as well as an ivermectin-resistant Cooperia oncophora population in cattle were studied. Experimentally infected sheep and cattle were used. Animals were treated orally, subcutaneously, or intravenously with cyclooctadepsipeptides. The anthelmintic effects were assessed by means of fecal egg count reductions and/or worm count reductions. Both, PF1022A and emodepside were found to be fully effective against these parasite populations. These findings confirm that this new class of compounds acts by a different mode of action compared to the above-mentioned anthelmintics.

Methylation analysis of the neurofibromatosis type 1 (NF1) promoter in peripheral nerve sheath tumours.

Peripheral nerve sheath tumours are hallmarks of neurofibromatosis type 1 (NF1). Development of plexiform neurofibromas to malignant peripheral nerve sheath tumours (MPNST) is common. The NF1 gene promoter harbours a hypomethylated CpG island. Thus, methylation changes may be involved in the development of different types of neurofibromas and malignant transformation. We investigated NF1-associated dermal (n=9) and plexiform neurofibromas (n=7), MPNST (n=5) and non-NF1 leucocyte samples (n=20) for their methylation pattern by bisulphite genomic sequencing. We could not find global hypermethylation in the NF1 promoter in our series. Nevertheless, site-specific methylation, involving transcription factor binding sites for SP1, CRE (-10), and AP-2, was observed. One region of the 5'-UTR (untranslated region) overlapping with a putative AP-2 binding site was methylated at 30-100% in 4/20 control samples. In conclusion, we did not find hypermethylation in NF1-associated tumours. Instead, low level methylation could parallel a global genomic hypomethylation in malignancy.