RASSF7 expression and its regulatory roles on apoptosis in human intervertebral disc degeneration

Apoptosis plays an important role in intervertebral disc degeneration (IDD). Overwhelming evidence indicates that RASSF7 is essential for cell growth and apoptosis. Recently, it has been noted that the JNK signaling can be negatively regulated by suppressing phosphorylated-MKK7 activation during pro-apoptosis. We aimed to investigate the RASSF7 expression level in human degenerative nucleus pulposus (NP) cells and non-degenerative NP cells and the link between RASSF7-JNK with NP cells apoptosis. We harvested NP tissues from 20 IDD patients as disease group and 8 cadaveric donors as normal controls. We detected RASSF7 expression by Real-time-PCR and western blotting. Consequently, we found that the expression of RASSF7 was higher in non-degenerative group than in degenerative group (P<0.05). Overexpression of RASSF7 in degenerative NP cells led to decreased apoptosis rate than that in scramble group (P<0.05). Collectively, our findings suggest that RASSF7 plays an important role in human IDD and RASSF7 might be potentially developed as a curative agent.

Placement Methods of Phosphorus and Potassium for Corn and Soybean Managed with No-till and Chisel-plow/Disk Tillage

No-till management limits the incorporation of crop residue and fertilizer with soil resulting in wetter, colder soils and the accumulation of organic matter, phosphorus (P), and potassium (K) near the soil surface. Banding of P and K could be more effective than broadcast fertilization by counteracting stratification, applying nutrients in the root zone (starter effect), and minimizing reactions with the soil that may reduce their availability to plants. Therefore, this long-term study was established in 1994 to evaluate P and K fertilizer placement methods and grain yield of corn-soybean rotations managed with notill and chisel-plow/disk tillage.

Secchi disk data collection for the North Sea and Baltic Sea

This paper presents the results of a Secchi depth data mining study for the North Sea - Baltic Sea region. 40,829 measurements of Secchi depth were compiled from the area as a result of this study. 4.3% of the observations were found in the international data centers [ICES Oceanographic Data Center in Denmark and the World Ocean Data Center A (WDC-A) in the USA], while 95.7% of the data was provided by individuals and ocean research institutions from the surrounding North Sea and Baltic Sea countries. Inquiries made at the World Ocean Data Center B (WDC-B) in Russia suggested that there could be significant additional holdings in that archive but, unfortunately, no data could be made available. The earliest Secchi depth measurement retrieved in this study dates back to 1902 for the Baltic Sea, while the bulk of the measurements were gathered after 1970. The spatial distribution of Secchi depth measurements in the North Sea is very uneven with surprisingly large sampling gaps in the Western North Sea. Quarterly and annual Secchi depth maps with a 0.5° x 0.5° spatial resolution are provided for the transition area between the North Sea and the Baltic Sea (4°E-16°E, 53°N-60°N).

Validation of a CFD wake model based on the actuator disk technique and the thrust coefficient. Preliminary results

A simplified CFD wake model based on the actuator-disk concept is used to simulate the wind turbine, represented by an actuator disk upon which a distribution of forces, defined as axial momentum sources, are applied on the incoming flow. The rotor is supposed to be uniformly loaded, with the exerted forces as a function of the incident wind speed, the thrust coefficient and the rotor diameter. The model is validated through experimental measurements downwind of a wind turbine in terms of wind speed deficit. Validation on turbulence intensity will also be made in the near future.

Stability of liquid bridges between an elliptical and a circular supporting disk

A numerical method has been developed to determine the stability limits for liquid bridges held between noncircular supporting disks and the application to a configuration with a circular and an elliptical disk subjected to axial acceleration has been made. The numerical method led to results very different from the available analytical solution which has been revisited and a better approximation has been obtained. It has been found that just retaining one more term in the asymptotic analysis the solution reproduces the real behavior of the configuration and the numerical results.

Disk halo size measured in individuals with monofocal versus diffractive multifocal intraocular lenses

PURPOSE: To compare disk halo size in response to a glare source in eyes with an aspheric apodized diffractive multifocal intraocular lens (IOL) or aspheric monofocal IOL. SETTING: Rementeria Ophthalmological Clinic, Madrid, Spain. DESIGN: Prospective randomized masked study. METHOD: Halo radius was measured using a vision monitor (MonCv3) with low-luminance optotypes in eyes that had cataract surgery and bilateral implantion of an Acrysof Restor SN6AD1 multifocal IOL or Acrysof IQ monofocal IOL 6 to 9 months previously. The visual angle subtended by the disk halo radius was calculated in minutes of arc (arcmin). Patient complaints of halo disturbances were recorded. Monocular uncorrected distance visual acutity (UDVA) and corrected distance visual acuity (CDVA) were measured using high-contrast (96%) and low-contrast (10%) logMAR letter charts. RESULTS: The study comprised 39 eyes of 39 subjects (aged 70 to 80 years); 21 eyes had a multifocal IOL and 18 eyes a monofocal IOL. The mean halo radius was 35 arcmin larger in the multifocal IOL group than the monofocal group (P<.05). Greater halo effects were reported in the multifocal IOL group (P<.05). The mean monocular high-contrast UDVA and low-contrast UDVA did not vary significantly between groups, whereas the mean monocular high-contrast CDVA and low-contrast CDVA were significantly worse at 0.12 logMAR and 0.13 logMAR in the multifocal than in the monofocal IOL group, respectively (P <.01). A significant positive correlation was detected by multiple linear regression between the halo radius and low-contrast UDVA in the multifocal IOL group (r = 0.72, P<.001). CONCLUSIONS: The diffractive multifocal IOL gave rise to a larger disk halo size, which was correlated with a worse low-contrast UDVA.

New galactic star clusters discovered in the VVV survey. Candidates projected on the inner disk and bulge

Context. VISTA Variables in the Vía Láctea (VVV) is one of six ESO Public Surveys using the 4 meter Visible and Infrared Survey Telescope for Astronomy (VISTA). The VVV survey covers the Milky Way bulge and an adjacent section of the disk, and one of the principal objectives is to search for new star clusters within previously unreachable obscured parts of the Galaxy. Aims. The primary motivation behind this work is to discover and analyze obscured star clusters in the direction of the inner Galactic disk and bulge. Methods. Regions of the inner disk and bulge covered by the VVV survey were visually inspected using composite JHKS color images to select new cluster candidates on the basis of apparent overdensities. DR1, DR2, CASU, and point spread function photometry of 10 × 10 arcmin fields centered on each candidate cluster were used to construct color–magnitude and color–color diagrams. Follow-up spectroscopy of the brightest members of several cluster candidates was obtained in order to clarify their nature. Results. We report the discovery of 58 new infrared cluster candidates. Fundamental parameters such as age, distance, and metallicity were determined for 20 of the most populous clusters.

The Gaia-ESO Survey: radial metallicity gradients and age-metallicity relation of stars in the Milky Way disk

We study the relationship between age, metallicity, and α-enhancement of FGK stars in the Galactic disk. The results are based upon the analysis of high-resolution UVES spectra from the Gaia-ESO large stellar survey. We explore the limitations of the observed dataset, i.e. the accuracy of stellar parameters and the selection effects that are caused by the photometric target preselection. We find that the colour and magnitude cuts in the survey suppress old metal-rich stars and young metal-poor stars. This suppression may be as high as 97% in some regions of the age-metallicity relationship. The dataset consists of 144 stars with a wide range of ages from 0.5 Gyr to 13.5 Gyr, Galactocentric distances from 6 kpcto 9.5 kpc, and vertical distances from the plane 0 < |Z| < 1.5 kpc. On this basis, we find that i) the observed age-metallicity relation is nearly flat in the range of ages between 0 Gyr and 8 Gyr; ii) at ages older than 9 Gyr, we see a decrease in [Fe/H] and a clear absence of metal-rich stars; this cannot be explained by the survey selection functions; iii) there is a significant scatter of [Fe/H] at any age; and iv) [Mg/Fe] increases with age, but the dispersion of [Mg/Fe] at ages >9 Gyr is not as small as advocated by some other studies. In agreement with earlier work, we find that radial abundance gradients change as a function of vertical distance from the plane. The [Mg/Fe] gradient steepens and becomes negative. In addition, we show that the inner disk is not only more α-rich compared to the outer disk, but also older, as traced independently by the ages and Mg abundances of stars.

Non-Viral Gene Therapy to Cells of the bovine Intervertebral Disc

Background. Low back pain is an increasing global health problem, which is associated with intervertebral disc (IVD) damage and degeneration. Major changes occur in the nucleus pulposus (NP), with the degradation of the extracellular matrix (ECM).1 Further studies showed that growth factors from transforming growth factor β (TGFβ) and bone morphogenic proteins (BMP) family may induce chondrogenic differentiation of mesenchymal stem cells (MSC).2 Focusing on non-viral gene therapies and their possible translation into the clinics, we investigated if GDF6 (syn. BMP13 or CDMP2) can induce regeneration of degraded NP. We hypothesized that IVD transfected with plasmid over-expressing GDF6 also up-regulates other NP- and chondrogenic cell markers and enhances ECM deposition. Methods. Bovine nucleus pulposus (bNPC) and annulus fibrosus cells (bAFC) were harvested from bovine coccygeal IVD. Primary cells were then electroporized with plasmid GDF6 (Origene, vector RG211366) by optimizing parameters using the Neon Transfection system (Life Technologies, Basel). After transfection, cells were cultured in 2D monolayer or 3D alginate beads for 7, 14 or 21 days. Transfection efficiency of pGDF6 was analyzed by immunohistochemistry and fluorescent microscopy. Cell phenotype was quantified by real-time RT-PCR. To test a non-viral gene therapy applied directly to 3D whole organ culture, coccygeal bovine IVDs were harvested as previously described. Bovine IVDs were transfected by injection of plasmid GDF6 into the center. Electroporation was performed with ECM830 Square Wave Electroporation System (Harvard Apparatus, MA) using 2-needle array electrode or tweezertrodes. 72 h after tranfection discs were fixed and cryosectioned and analyzed by immunofluorescence against GDF6. Results. RT-PCR and immunohistochemistry confirmed up-regulation of GFP and GDF6 in the primary bNPC/bAFC culture. The GFP-tagged GDF6 protein, however, was not visible, possibly due to failure of dimer formation as a result of fusion structure. Organ IVD culture transfection revealed GDF6 positive staining in the center of the disc using 2-needle array electrode. Results from tweezertrodes did not show any GDF6 positive cells. Conclusion. Non-viral transfection is an appealing approach for gene therapy as it fulfills the translational safety aspects of transiency and lacks the toxic effects of viral transduction. We identified novel parameters to successfully transfect primary bovine IVD cells. For transfection of whole IVD explants electroporation parameters need to be further optimized. Acknowledgements. This project was funded by the Lindenhof Foundation (Funds “Research & Teaching”) Project no. 13-02-F. The imaging part of this study was performed with the facility of the Microscopy Imaging Center (MIC), University of Bern. References. Roughly PJ (2004): Spine (Phila), 29:2691-2699 Clarke LE, McConell JC, Sherratt MJ, Derby B, Richardson SM, Hoyland JA (2014), Arthritis Research & Therapy, 16:R67

Compression Loading Induced Cellular Stress Response of Intervertebral Disc Cells in Organ Culture

Introduction: Mechanical stress is often associated to interverterbal disc (IVD) degeneration and the effect of mechanical loading on IVD has been studied and reviewed.1,2 Previously, expression of heat shock proteins, HSP70 and HSP27 has been found in pathological discs.3 However, there is no direct evidence on whether IVD cells respond to the mechanical loading by expression of HSPs. The objective of this study is to investigate the stress response of IVD cells during compressive loading in an organ culture. Materials and Methods: Fresh adult bovine caudal discs were cultured with compressive loading applied at physiological range. Effect of loading type (static and dynamic) and repeated loading (2 hours per day for 2 days) were studied. Nucleus pulposus (NP) and annulus fibrosus (AF) of the IVD were retrieved at different time points: right after loading and right after resting. Positive control discs were heat shocked (43°C). Cell activity was assessed and expression of stress response genes (HSP70 and HSF1) and matrix remodeling genes (ACAN, COL2, COL1, ADAMTS4, MMP3 and MMP13) were studied. Results: Cell activity was maintained in all groups. Both NP and AF expressed high level of HSP70 in heat shock groups, confirming their expression in response to stress. In NP, expression of HSP70 was up-regulated after static loading and dynamic loading with higher fold change was observed after static loading. During repeated loading, HSP70 appeared to be upregulated right after loading and decreased after resting. Such trend was not observed in AF and HSF1 levels. Expressions of matrix remodeling genes did not change significantly with loading except ADAMTS4 decreased in AF during static loading. Conclusion: This study demonstrated that NP cells upregulate expression of HSP70 in response to loading induced stress without changing cell activity and matrix remodeling significantly. Acknowledgments: This project was funded by AO Spine (AOSPN) (grant number: SRN_2011_14) and a fellowship exchange award by AO Spine Scientific Research Network (SRN).