A note on integumental (1-->3)(1-->6)beta-D-glucan permeation, using the porcine ear skin model

Based on carbohydrate histochemical methods and the porcine ear skin model, the study demonstrates the rapid permeation of (1-->3)(1-->6)beta-d-glucans from a cosmetic formulation into the mammalian epidermis.     

Intra-oral compartment pressures: a biofunctional model and experimental measurements under different conditions of posture

Oral posture is considered to have a major influence on the development and reoccurrence of malocclusion. A biofunctional model was tested with the null hypotheses that (1) there are no significant differences between pressures during different oral functions and (2) between pressure measurements in different oral compartments in order to substantiate various postural conditions at rest by intra-oral pressure dynamics. Atmospheric pressure monitoring was simultaneously carried out with a digital manometer in the vestibular inter-occlusal space (IOS) and at the palatal vault (sub-palatal space, SPS). Twenty subjects with normal occlusion were evaluated during the open-mouth condition (OC), gently closed lips (semi-open compartment condition, SC), with closed compartments after the generation of a negative pressure (CCN) and swallowing (SW). Pressure curve characteristics were compared between the different measurement phases (OC, SC, CCN, SW) as well as between the two compartments (IOS, SPS) using analysis of variance and Wilcoxon matched-pairs tests adopting a significance level of α = 0.05. Both null hypotheses were rejected. Average pressures (IOS, SPS) in the experimental phases were 0.0, −0.08 (OC); −0.16, −1.0 (SC); −48.79, −81.86 (CCN); and −29.25, −62.51 (SW) mbar. CCN plateau and peak characteristics significantly differed between the two compartments SPS and IOS. These results indicate the formation of two different intra-oral functional anatomical compartments which provide a deeper understanding of orofacial biofunctions and explain previous observations of negative intra-oral pressures at rest.     

Zink finger nucleases and siRNAs: use in transgenic pig production for xenotransplantation

Emerging technologies, including the use of specific nucleases, transposons and specific siRNAs will bring significant improvements with regard to efficiency, precision and safety of gene transfer. Here, we summarize recent findings from our laboratory on the integration of Zinc‐finger nuclease technology (ZFNs) and specific siRNAs into a research programme towards the development and characterization of multi‐transgenic pigs suitable for xenotransplantation. Zinc‐finger nucleases (ZFNs) are powerful tools for producing gene knockouts (KOs) with very high efficiency. ZFNs were used to induce a biallelic knockout of the porcine α1,3‐galactosyltransferase (GGTA1) gene by employing primary porcine fibroblasts that were treated with ZFNs designed against the region coding for the catalytic core of GGTA1 in SCNT. This resulted in biallelic knockout of a high proportion of ZFN‐treated cells. ZFN‐mediated genetic modification did not interfere with the cloning process. Off‐target cleavage events or integration of the ZFN‐coding plasmid were not detected. The absence of Gal epitopes was determined by FACS and the GGTA1‐KO phenotype was confirmed by a complement lysis assay that demonstrated significant protection of GGTA1‐KO fibroblasts relative to wild‐type cells. The ZFNs were functional irrespective of the sex of the donor cells, thus allowing the effective production of female and male homozygous Gal kO pigs via somatic cloning and the rapid establishment of Gal‐/‐ pig lines. These pigs will serve as genetic background upon which additional transgenes can be inserted and expressed. SiRNAs have emerged as useful tool to knockdown specific genes in complex organisms for a variety of targets and serve as an important tool in the functional analysis of specific genes. The acute vascular rejection (AVR) remains the main hurdle for long term survival of a porcine xenograft after transplantation into primates. Immunological reactions and molecular incompatibilities can lead to endothelial activation and microvascular thrombosis. Knock‐down of tissue factor (TF) could be a promising mechanism to prevent AVR. TF is a key molecule of the extrinsic coagulation pathway. It functions as cell surface receptor for coagulation factor VIIa and thereby initiates thrombin formation. Since TF knock‐out was lethal in the mouse model, we tested different siRNAs in their efficiency in a porcine cell line to knock‐down TF. Two siRNAs reduced the mRNA level of TF to <3% compared with wild type controls as determined by RT‐PCR. Subsequently, porcine fetal fibroblasts (PFF) were co‐transfected with constructs coding for one of the promising siRNAs and a DsRed vector which confers neomycin resistance and red fluorescence. Transfected cells were selected with neomycin for 14 days and observed for the occurrence of fluorescence. Integration of the siRNA vector was confirmed by PCR. Cell clones positive for TF knock‐down siRNA served as donor cells for somatic cell nuclear transfer (SCNT). Reconstructed embryos were transferred to synchronised recipient sows. Eight transfers resulted in four pregnancies. One foster animal was sacrificed on day 35 of gestation. Three fetuses were recovered that had integrated the TF knock‐down siRNA and the derived fetal cells were used for recloning and more piglets were produced. The functionality of the knock‐down is tested in a coagulation assay and in an established TNF alpha induced assay using cultured endothelial cells of knock‐down piglets in comparison with wild type counterparts. Exposure to inflammation mediators is known stimulate expression of TF and adhesion molecules in the endothelium. Our genetically modified TF transgenic pigs will be further characterized and pigs with the desired TF knockdown will combined with already existing genetic modifications to generate multi‐transgenic pigs. This model would be of great importance to prolong xenograft survival and move porcine xenografts to the pre‐clinical level. This project was funded by DFG (Ni 256/22‐3 u. 4) and the EU project “Xenome” (LSHB‐CT‐2006‐037377).     

Phase I Dose-Escalation Study of the Safety, Pharmacokinetics, and Pharmacodynamics of the MEK Inhibitor RO4987655 (CH4987655) in Patients with Advanced Solid Tumors

PURPOSE: This phase I study of the mitogen-activated protein/extracellular signal-regulated kinase inhibitor RO4987655 (CH4987655) assessed its maximum tolerated dose (MTD), dose-limiting toxicities (DLT), safety, pharmacokinetic/pharmacodynamic profile, and antitumor activity in patients with advanced solid tumors. Patients and Methods: An initial dose escalation was conducted using a once-daily dosing schedule, with oral RO4987655 administered at doses of 1.0 to 2.5 mg once daily over 28 consecutive days in 4-week cycles. Doses were then escalated from 3.0 to 21.0 mg [total daily dose (TDD)] using a twice-daily dosing schedule. RESULTS: Forty-nine patients were enrolled. DLTs were blurred vision (n = 1) and elevated creatine phosphokinase (n = 3). The MTD was 8.5 mg twice daily (TDD, 17.0 mg). Rash-related toxicity (91.8%) and gastrointestinal disorders (69.4%) were the most frequent adverse events. The pharmacokinetic profile of RO4987655 showed dose linearity and a half-life of approximately 4 hours. At the MTD, target inhibition, assessed by suppression of extracellular signal-regulated kinase phosphorylation in peripheral blood mononuclear cells, was high (mean 75%) and sustained (90% of time >IC(50)). Of the patients evaluable for response, clinical benefit was seen in 21.1%, including two partial responses (one confirmed and one unconfirmed). 79.4% of patients showed a reduction in fluorodeoxyglucose uptake by positron emission tomography between baseline and day 15. CONCLUSION: In this population of heavily pretreated patients, oral RO4987655 showed manageable toxicity, a favorable pharmacokinetics/pharmacodynamics profile, and promising preliminary antitumor activity, which has been further investigated in specific populations of patients with RAS and/or RAF mutation driven tumors. Clin Cancer Res; 18(17); 4794-805. ?2012 AACR.     

Zur Wirkung der Kalksalze Auf die Blutgerinnung Bei Oraler und Intravenöser Zufuhr

Ohne Zusammenfassung     

über Kollargolbehandlung des Chronischen Gelenkrheumatismus

Ohne Zusammenfassung     

Rehospitalization in the first 2 years of life in children born preterm

Aim: Aim of the study is to investigate the frequency of and predictors for rehospitalization within the first 2 years of life among preterm infants. Methods: All children born before 32 weeks of gestation in Northern Tyrol between January 2003 and July 2008 were prospectively enrolled. Data on rehospitalizations were obtained from hospital admission records. The association between candidate risk factors and readmission was analysed by means of logistic regression analysis. Results: In the first and second years of life, 151 and 93 of 377 children (40.1% and 24.7%), respectively, were readmitted to one of the hospitals in Northern Tyrol. The most common causes of rehospitalization were respiratory disorders, accounting for 42.1% and 47.4% of total readmissions in the first and second years of life. Chronic lung disease (CLD), male sex and smoking in pregnancy were risk conditions relevant to readmission in the first year of life, but only CLD in the second year. Conclusion: Infants born before 32 weeks of gestation have a high risk of rehospitalization with respiratory illness significantly contributing to postdischarge morbidity. Neonatal intensive care should aim to further improve respiratory health in preterm infants, and adequate follow‐up services must be offered.