Bmp signaling regulates a dose-dependent transcriptional program to control facial skeletal development

We performed an in depth analysis of Bmp4, a critical regulator of development, disease, and evolution, in cranial neural crest (CNC). Conditional Bmp4 overexpression, using a tetracycline-regulated Bmp4 gain-of-function allele, resulted in facial skeletal changes that were most dramatic after an E10.5 Bmp4 induction. Expression profiling uncovered a signature of Bmp4-induced genes (BIG) composed predominantly of transcriptional regulators that control self-renewal, osteoblast differentiation and negative Bmp autoregulation. The complimentary experiment, CNC inactivation of Bmp2, Bmp4 and Bmp7, resulted in complete or partial loss of multiple CNC-derived skeletal elements, revealing a crucial requirement for Bmp signaling in membranous bone and cartilage development. Importantly, the BIG signature was reduced in Bmp loss-of-function mutants, indicating Bmp-regulated target genes are modulated by Bmp dose. Chromatin immunoprecipitation (ChIP) revealed a subset of the BIG signature, including Satb2, Smad6, Hand1, Gadd45γ and Gata3, that was bound by Smad1/5 in the developing mandible, revealing direct Smad-mediated regulation. These data support the hypothesis that Bmp signaling regulates craniofacial skeletal development by balancing self-renewal and differentiation pathways in CNC progenitors

Bmp signaling represses Vegfa to promote outflow tract cushion development

Congenital heart disease (CHD) is a devastating anomaly that affects ∼1% of live births. Defects of the outflow tract (OFT) make up a large percentage of human CHD. We investigated Bmp signaling in mouse OFT development by conditionally deleting both Bmp4 and Bmp7 in the second heart field (SHF). SHF Bmp4/7 deficiency resulted in defective epithelial to mesenchymal transition (EMT) and reduced cardiac neural crest ingress, with resultant persistent truncus arteriosus. Using a candidate gene approach, we found that Vegfa was upregulated in the Bmp4/7 mutant hearts. To determine if Vegfa is a downstream Bmp effector during EMT, we examined whether Vegfa is transcriptionally regulated by the Bmp receptor-regulated Smad. Our findings indicate that Smad directly binds to Vegfa chromatin and represses Vegfa transcriptional activity. We also found that Vegfa is a direct target for the miR-17-92 cluster, which is also regulated by Bmp signaling in the SHF. Deletion of miR-17-92 reveals similar phenotypes to Bmp4/7 SHF deletion. To directly address the function of Vegfa repression in Bmp-mediated EMT, we performed ex vivo explant cultures from Bmp4/7 and miR-17-92 mutant hearts. EMT was defective in explants from the Bmp4/7 double conditional knockout (dCKO; Mef2c-Cre;Bmp4/7(f/f)) and miR-17-92 null. By antagonizing Vegfa activity in explants, EMT was rescued in Bmp4/7 dCKO and miR-17-92 null culture. Moreover, overexpression of miR-17-92 partially suppressed the EMT defect in Bmp4/7 mutant embryos. Our study reveals that Vegfa levels in the OFT are tightly controlled by Smad- and microRNA-dependent pathways to modulate OFT development

Forced expression of the cell cycle inhibitor p57Kip2 in cardiomyocytes attenuates ischemia-reperfusion injury in the mouse heart

<p>Abstract</p> <p>Background</p> <p>Myocardial hypoxic-ischemic injury is the cause of significant morbidity and mortality worldwide. The cardiomyocyte response to hypoxic-ischemic injury is known to include changes in cell cycle regulators. The cyclin-dependent kinase inhibitor <it>p57</it>^{<it>Kip</it>2}is involved in cell cycle control, differentiation, stress signaling and apoptosis. In contrast to other cyclin-dependent kinase inhibitors, p57^Kip2expression diminishes during postnatal life and is reactivated in the adult heart under conditions of cardiac stress. Overexpression of <it>p57</it>^{<it>Kip</it>2}has been previously shown to prevent apoptotic cell death <it>in vitro </it>by inhibiting stress-activated kinases. Therefore, we hypothesized that <it>p57</it>^{<it>Kip</it>2}has a protective role in cardiomyocytes under hypoxic conditions. To investigate this hypothesis, we created a transgenic mouse (<it>R26loxpTA-p57</it>^<it>k</it>/+) that expresses p57^Kip2specifically in cardiac tissue under the ventricular cardiomyocyte promoter <it>Mlc2v</it>.</p> <p>Results</p> <p>Transgenic mice with cardiac specific overexpression of <it>p57</it>^{<it>Kip</it>2}are viable, fertile and normally active and their hearts are morphologically indistinguishable from the control hearts and have similar heart weight/body weight ratio. The baseline functional parameters, including left ventricular systolic pressure (LVSP), left ventricular end diastolic pressure (LVEDP), LVdp/dt_max, heart rate (HR) and rate pressure product (RPR) were not significantly different between the different groups as assessed by the Langendorff perfused heart preparation. However, after subjecting the heart <it>ex vivo </it>to 30 minutes of ischemia-reperfusion injury, the <it>p57</it>^{<it>Kip</it>2}overexpressing hearts demonstrated preserved cardiac function compared to control mice with higher left ventricular developed pressure (63 ± 15 vs 30 ± 6 mmHg, p = 0.05), rate pressure product (22.8 ± 4.86 vs 10.4 ± 2.1 × 10³bpm × mmHg, p < 0.05) and coronary flow (3.5 ± 0.5 vs 2.38 ± 0.24 ml/min, p <0.05).</p> <p>Conclusion</p> <p>These data suggest that forced cardiac expression of p57^Kip2does not affect myocardial growth, differentiation and baseline function but attenuates injury from ischemia-reperfusion in the adult mouse heart.</p

The relationship between built-up areas and the spatial development of the mean maximum urban heat island in Debrecen, Hungary

The climate of built-up regions differs significantly from rural regions and the most important modifying effect of urbanization on local climate is the urban temperature excess, otherwise called the urban heat island (UHI). This study examines the influence of built-up areas on the near-surface air temperature field in the case of the medium-sized city of Debrecen, Hungary. Mobile measurements were used under different weather conditions between March 2002 and March 2003. Efforts concentrated on the determination of the spatial distribution of mean maximum UHI intensity with special regard to land-use features such as built-up ratio and its areal extensions. In both (heating and non-heating) seasons the spatial distribution of the UHI intensity field showed a basically concentric shape with local anomalies. The mean maximum UHI intensity reaches more than 2.0 degrees C (heating season) and 2.5 degrees C (non-heating season) in the centre of the city. We established the relationship between the above-mentioned land-use parameters and mean maximum UHI intensity by means of multiple linear regression analysis. As the measured and predicted mean maximum UHI intensity patterns show, there is an obvious connection between the spatial distribution of urban thermal excess and the land-use parameters examined, so these parameters play a significant role in the development of the strong, UHI intensity field over the city. Copyright (c) 2005 Royal Meteorological Society

Long-term outcome data from 121 patients treated with Gamma Knife stereotactic radiosurgery as salvage therapy for focally recurrent high-grade gliomas.

Introduction: We examined patient outcomes after Gamma Knife stereotactic radiosurgery (GKSRS) salvage therapy for recurrent high-grade gliomas (HGGs) to determine whether tumor grade or lesion size affected overall survival (OS) and progression-free survival (PFS). Methods: This single-center retrospective study assessed radiographic response and clinical outcomes following GKSRS salvage treatment of recurrent malignant gliomas (January 2005-March 2014). Results: A total of 121 patients (67 female) with 132 tumors were treated. Median (range) PFS was 4.7 (3.9-5.4) months for the cohort, 6.8 (4.6-8.9) months for initial grade 2 tumors, 4.2 (1.9-6.5) months for initial grade 3 tumors, and 4.3 (3.7-4.9) months for initial grade 4 tumors. Patients with small lesions (≤6.7 cm Conclusions: GKSRS offers meaningful salvage therapy with minimal morbidity in appropriately selected patients with focally recurrent HGGs

Local-scale urban meteorological parameterization scheme (LUMPS): longwave radiation parameterization and seasonality-related developments

Recent developments to the Local-scale Urban Meteorological Parameterization Scheme (LUMPS), a simple model able to simulate the urban energy balance, are presented. The major development is the coupling of LUMPS to the Net All-Wave Radiation Parameterization (NARP). Other enhancements include that the model now accounts for the changing availability of water at the surface, seasonal variations of active vegetation, and the anthropogenic heat flux, while maintaining the need for only commonly available meteorological observations and basic surface characteristics. The incoming component of the longwave radiation (L↓) in NARP is improved through a simple relation derived using cloud cover observations from a ceilometer collected in central London, England. The new L↓ formulation is evaluated with two independent multiyear datasets (Łódź, Poland, and Baltimore, Maryland) and compared with alternatives that include the original NARP and a simpler one using the National Climatic Data Center cloud observation database as input. The performance for the surface energy balance fluxes is assessed using a 2-yr dataset (Łódź). Results have an overall RMSE < 34 W m−2 for all surface energy balance fluxes over the 2-yr period whe

Omeprazole Increases the Efficacy of Acyclovir Against Herpes Simplex Virus Type 1 and 2

Omeprazole was shown to improve the anti-cancer effects of the nucleoside analogue 5-fluorouracil. Here, we combined omeprazole with the antiviral nucleoside analogues ribavirin and acyclovir. Omeprazole did not affect the antiviral effects of ribavirin in non-toxic concentrations up to 80 μg/mL but increased the acyclovir-mediated effects on herpes simplex virus 1 and 2 (HSV-1 and -2) replication in a dose-dependent manner. Omeprazole alone reduced HSV-1 and -2 titers [but not HSV-induced formation of cytopathogenic effects (CPE)] at concentrations ≥40 μg/mL. However, it exerted substantially stronger effects on acyclovir activity and also increased acyclovir activity at lower concentrations that did not directly interfere with HSV replication. Omeprazole 80 μg/mL caused a 10.8-fold (Vero cells) and 47.7-fold (HaCaT cells) decrease of the acyclovir concentrations that reduced HSV-1-induced CPE formation by 50% (IC50). In HSV-2-infected cells, omeprazole 80 μg/mL reduced the acyclovir IC50 by 7.3- (Vero cells) and 12.9-fold (HaCaT cells). In HaCaT cells, omeprazole 80 μg/mL reduced the HSV-1 titer in the presence of acyclovir 1 μg/mL by 1.6 × 105-fold and the HSV-2 titer in the presence of acyclovir 2 μg/mL by 9.2 × 103-fold. The proton pump inhibitors pantoprazole, rabeprazole, lansoprazole, and dexlansoprazole increased the antiviral effects of acyclovir in a similar fashion as omeprazole, indicating this to be a drug class effect. In conclusion, proton pump inhibitors increase the anti-HSV activity of acyclovir and are candidates for antiviral therapies in combination with acyclovir, in particular for topical preparations for the treatment of immunocompromised individuals who are more likely to suffer from severe complications

Absence of RKIP expression is an independent prognostic biomarker for gastric cancer patients

Gastric cancer is a leading cause of cancer-related mortality, and the presence of lymph node metastasis an important prognostic factor. Downregulation of RKIP has been associated with tumor progression and metastasis in several types of neoplasms, being currently categorized as a metastasis suppressor gene. Our aim was to determine the expression levels of RKIP in gastric tissues and to evaluate its impact in the clinical outcome of gastric carcinoma patients. RKIP expression levels were studied by immunohistochemistry in a series of gastric tissues. Overall, we analysed 222 non-neoplastic gastric tissues, 152 primary tumors and 42 lymph node metastasis samples. We observed that RKIP was highly expressed in ~83% of non-neoplastic tissues (including normal tissue and metaplasia), was lost in ~56% of primary tumors and in ~90% of lymph node metastasis samples. Loss of RKIP expression was significantly associated with several markers of poor clinical outcome, including the presence of lymph node metastasis. Furthermore, the absence of RKIP protein constitutes an independent prognostic marker for these patients. In conclusion, RKIP expression is significantly lost during gastric carcinoma progression being almost absent in lymph node metastasis samples. Of note, we showed that the absence of RKIP expression is associated with poor outcome features of gastric cancer patients, this being also an independent prognostic marker.Olga Martinho was recipient of a PhD fellowship (SFRH/BD/36463/2007) from Fundacao para a Ciencia e Tecnologia (FCT), Portugal

Assessing the impact of changes in surface cover, human behaviour and climate on energy partitioning across Greater London

Climate-sensitive urban design is an increasingly important consideration for city planners and policy makers. This study demonstrates the use of a biophysical model to assess the response of urban climate to various changes, including population growth, reduced energy use, urban development and urban greening initiatives. Model inputs are intentionally derived using only publicly available information and assumptions involved in collating the data are discussed. Results are summarised in terms of the energy partitioning which captures changes in meteorology, surface characteristics and human behaviour. The model has been recently evaluated for the region, and those findings are drawn upon here to discuss the model’s capabilities and limitations. Model simulations demonstrate how both intentional and inadvertent changes to the urban landscape can alter the urban climate. For example, the impact of population growth depends on where, and how, people are housed, and recent changes in garden composition have reduced evaporation. This study has been designed so that model output could be combined with socio-economic data in future, enabling both risk and vulnerability to be considered together