text:
|By Donna Parent
Article Rating:
|November 2, 2011 06:00 AM EDT
Enterprise CFOs are taking an increasingly active role in managing the IT decision making process. As a result, IT professionals need to be prepared to quickly show how innovative technology investments equate to real business value. According to a recent industry report, only 5 percent of CIOs have the power to authorize IT investments. That same study reports that IT departments are now, more than ever, reporting directly to the CFO (42 percent) as opposed to the 33 percent reporting to the CEO. A company can't stay competitive and productive without the right tools, technology and infrastructure. And, today's CIOs and IT leaders need the approval of the finance department, which means technology investments need to be more tightly coupled and aligned to company goals and business value.
It has also become increasingly important to prove how new investments can drive additional ROI from the company's existing infrastructure, reduce overall business costs, optimize business processes and improve business productivity, while also demonstrating overall interoperability with existing and future IT investments. Common questions finance asks IT are: "Why do we need another XYZ tool when we already have several throughout the enterprise?" or "How will this product add value to the business and when?"
Making the Case for End User Experience Management
When it comes to application performance management (APM) and end user experience, enterprises could have anywhere from five to over 100 management tools that they use in tandem to help manage their IT services. However, these tools are typically data center-centric - they are unable to monitor, detect and pinpoint the cause of application performance problems from the desktop vantage point. So despite the fact there's been a substantial investment in these APM tools, they cannot provide IT with visibility into understanding how end users are actually experiencing the IT services they consume. Therefore, IT is unable to proactively manage and address performance issues before business productivity is impacted.
Below are some suggested ways for IT to bridge the ROI use case gap and how to describe the real business value that can be delivered with the adoption of real end user experience monitoring and management tools, especially those focused on the desktop vantage point - which, by the way, is becoming more widely recognized as a necessity in order to fuel a true user-centric approach to proactive IT management.
According to Will Cappelli, research vice president of Gartner, Inc., "Technologies are required which will be able to penetrate the increasingly complex and opaque Internet edge and the correlation between edge events and host processes. In fact, we believe that such technologies that gather and analyze data from the point where users and customers actually access the data could, in some cases, replace APM technologies that are reliant on data-center-bound instrumentation points."
1. Define and Communicate the Limitations of Existing Products
Industry-leading analysts have established that in 74 percent of the reported help desk cases, IT first learns about performance and availability problems when users call the help desk. That's because existing application performance management products are data center-focused and provide very little visibility into real end user experience. As mentioned earlier, the average company has five APM products in place, many have as many as 25 different products, and 100+ is not uncommon. While each of these products may be critical for managing certain components of the IT infrastructure, they do not address the effective management of the end user experience, thereby curtailing both user and business productivity.
The recently published Gartner Magic Quadrant for User Application Performance Monitoring (by Will Cappelli and Jonah Kowall, September 19, 2011) has identified end user experience monitoring as one of the central components to the application performance management process because it "is precisely where business process execution interfaces with the IT stack; any monitoring effort that fails to capture the end user's experience is ultimately blind at the most direct point of encounter between IT and the business."
The problem seems to be a lack of the right tools - those that support business optimization, improvements to IT processes, and increased user productivity. User-centric, proactive IT management of the end user experience addresses this critical problem - the visibility gap - at its core.
2. Share How Current and Future IT Investments Impact Real End User Experience and Business Productivity
In the application world, companies make continued investments to improve application adoption and how they perform for their end users. And it's no secret that when key business applications aren't performing optimally, the impact can be felt both on the business and on corporate morale. According to analyst firm Enterprise Management Associates, large companies report that "downtime can cost in excess of $15,000 per minute for technology-dependent organizations, as applications drive revenue, productivity and brand value."
When precise insights are available into how users utilize their applications, real end user performance data can be leveraged to increase the ROI on these investments. For example, if IT management can validate that users are actively adopting and successfully using the application and their productivity is meeting expectations, this insight can eliminate the unnecessary and costly investment of additional licenses or "premium" packages based on usage. It can also help highlight the need for improved training for increased productivity and even reduce the deadly "shelf ware" sin, where 57 percent of global enterprises own more software licenses than actually deployed.
3. Be Prepared with Empirical Evidence to Prove Your Case
Desktop and infrastructure investments are critical to any IT organization. It is a challenge to keep up with the plethora of devices, protocols and innovations to understand which is better for your unique business and IT environment. The cost of selecting, purchasing and deploying new infrastructure technologies can be exorbitant. On top of that, you enter very risky waters if you lack the empirical evidence that the performance improvements will actually improve the end user experience.
Understanding the potential impact of software and application upgrades on end user experience is critical for the enterprise, as a single vulnerability can severely impact business continuity. The same applies to testing configuration management of infrastructure components and various operating systems. Comprehensive before-and-after comparisons validate performance and functionality prior to rolling out the application enterprise-wide. Visibility into the actual end user experience and performance that proves proposed changes substantially impact performance (or not) ensure that the investment decisions are based on empirical evidence versus assumption.
4. Evangelize the "Visibility Gap" and Reveal How It Threatens the Business
To understand the impact real end user experience has on business productivity, and ultimately the return on your existing and future IT investments, you must be able to measure and monitor the three primary components that dynamically interact to constantly impact how end users experience the IT services they consume in real-time. Otherwise, decisions will be made with a significant "visibility gap" in performance knowledge that can lead to unnecessary and costly missteps.
The first component is application performance. Latency, response time and end-to-end transaction time are all key elements in both the experience and measurement of application performance.
The second component is device performance. Even if the end-to-end transaction time of a particular application is excellent, if the underlying platform is sluggish - perhaps due to CPU power, memory availability or other background processes that are resource hogs - the end user's experience with what should be a high-performing application will be poor.
Ultimately, businesses deliver applications to enhance the productivity of end users, making user productivity the third component of end user experience. How many trades, calls or emails can the end user complete using a particular application running on a specific desktop platform? Productivity is impacted by error messages, non-responding and crashed applications, boot time, and the actual usability of the application.
Measuring and monitoring the three primary components (application, device and user) that dynamically interact to constantly impact end users experience in real-time is critical in order to close the "visibility gap" between how IT believes services are being consumed and how they are actually being experienced by the frontline.
5. Comprehensive Visibility Delivers Rapid Response and Cost Reduction
More often than not, the first and only indication of a problem on the frontline is when end users call the help desk - if they call at all - and by this time business and end user productivity has already been disrupted. Moreover, when users start calling it is very difficult for them to accurately describe the problems they are experiencing, while at the same time IT can't determine whether the issues are isolated or endemic.
Immediate awareness and rapid response to end user problems before end users call the help desk can reduce the costs associated with the need for manual monitoring and dedicated help desk/IT operations resources for severity type 1 and 2 problems. The ability to monitor the "Key-to-Glass" for any business activity running on any desktop (type), autonomic performance profiling and proactive incident detection all help contribute to cost reduction.
Today's CFO is "increasingly becoming the top technology investment decision maker in many organizations," but even though one report discussed earlier indicates this shift, another report, CIO magazine's 2010 State of the CIO, found that 43 percent of CIOs still report to CEOs, and just 19 percent report to CFOs. Whether the number is 19 percent or 42 percent, the more important takeaway is that CIOs and IT leaders are indeed increasing the communication path and their transparency with financial leaders.
To do so, CIOs are demonstrating their understanding of the direct impact IT tools and technologies have on improving business performance by delivering effective control, efficiency and business insight. They are making this crystal clear to their financial leader counterpart(s) and therefore bridging the gap between the enigma of IT and the real benefits and ROI being witnessed on an ongoing basis. This is how CIOs and CFOs will achieve common business goals. Gaining true end user experience management and monitoring is just one example of how an IT investment can be pivotal in driving more ROI from existing and planned IT infrastructure investments - something that everyone can rally behind.
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Photography in the National Parks: DOF, Foreground Objects and Framing
How and where you choose to focus on your image can make a huge difference in how a viewer perceives your national park photo. Rebecca Latson provides some tips for helping you do just that.
- By Rebecca Latson ... on June 24th, 2014

subset: commoncrawl


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DFI LANParty LT X48-T2R Motherboard
Version and / or Patch Used: 1.2
Timedemo or Level Used: Hardware OC Demo
Developer Homepage: http://www.humanhead.com
Product Homepage: http://www.prey.com
Prey is one of the newest games to be added to our benchmark line-up. It is based off the Doom 3 engine and offers stunning graphics passing what we've seen in Quake 4 and does put quite a lot of strain on our test systems.
Moving to the real world we don't see much difference between the two boards, even at overclocked speeds.
PRICING: You can find products similar to this one for sale below.
United States: Find other tech and computer products like this over at Amazon's website.
United Kingdom: Find other tech and computer products like this over at Amazon UK's website.
Canada: Find other tech and computer products like this over at Amazon Canada's website.
- Page 1 [Introduction]
- Page 2 [Specifications]
- Page 3 [The Box and What's Inside]
- Page 4 [The Motherboard]
- Page 5 [BIOS and Overclocking]
- Page 6 [Test System Setup and Memory Performance]
- Page 7 [Benchmarks - PCMark05]
- Page 8 [Benchmarks - Adobe Premiere Elements 3.0]
- Page 9 [Benchmarks - HDD Performance]
- Page 10 [Benchmarks - 3DMark06]
- Page 11 [Benchmarks - Prey]
- Page 12 [Benchmarks - Battlefield 2142]
- Page 13 [Benchmarks - Far Cry]
- Page 14 [Power Consumption Tests]
- Page 15 [Final Thoughts]
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Otsego comes together to support Davis family
Val Davis has been called the biggest cheerleader for Otsego athletics.
On Saturday, March 12, some of the student-athletes Davis has cheered for returned the favor.
Val Davis has been called the biggest cheerleader for Otsego athletics.
On Saturday, March 12, some of the student-athletes Davis has cheered for returned the favor.
Hopkins boys' basketball coach was under no illusions.
All good things must eventually come to an end.
That included the Otsego girls' basketball team's winning streak that dated back more than two months.
A pair of Allegan County schools cracked the top 10 at the Division 3 boys' swim and dive state finals at Eastern Michigan University on Saturday, March 12.
While one building at the entrance to downtown was marked for demolition another was leased to a new tenant during the Allegan City Council meeting Monday, March 14.
GANGES TWP.—Police say a man poured acid on a woman during a domestic assault early Wednesday, March 16.
Marilyn Maass, 89, passed away on Feb. 24, 2016, at Rice Estate in Columbia, S.C. She was born Marilyn Jean Leitz in Hastings on March 8, 1926, the daughter of the late Fred and Esther Leitz.
James J. Benes, 86, of Darien, Ill., died Feb. 7, 2016.
Susan Tomlinson, 70, of Douglas, passed away Tuesday, March 8, 2016, at home. She was born in Manchester, England.
William A. Manthey, 77, of Fennville passed away on Sunday, March 13, 2016, at Freedom Village in Holland. Born Aug.
Kaechele Publications, Inc.
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Phone: 269-673-5534
Fax: 269-673-5535
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The authors have no conflict of interest.
Research Article
BMP-2 Controls Alkaline Phosphatase Expression and Osteoblast Mineralization by a Wnt Autocrine Loop†
Article first published online: 1 OCT 2003
DOI: 10.1359/jbmr.2003.18.10.1842
Copyright © 2003 ASBMR
Journal of Bone and Mineral Research
Volume 18, Issue 10, pages 1842–1853, October 2003
Additional Information
How to Cite
Rawadi, G., Vayssière, B., Dunn, F., Baron, R. and Roman-Roman, S. (2003), BMP-2 Controls Alkaline Phosphatase Expression and Osteoblast Mineralization by a Wnt Autocrine Loop. J Bone Miner Res, 18: 1842–1853. doi: 10.1359/jbmr.2003.18.10.1842
- †
Publication History
- Issue published online: 2 DEC 2009
- Article first published online: 1 OCT 2003
- Manuscript Accepted: 4 JUN 2003
- Manuscript Revised: 18 APR 2003
- Manuscript Received: 24 JAN 2003
- Abstract
- Article
- References
- Cited By
- osteoblast;
- Wnt;
- β-catenin;
- alkaline phosphatase
Wnt/β-catenin signaling has recently been suggested to be involved in bone biology. The precise role of this cascade in osteoblast differentiation was examined. We show that a Wnt autocrine loop mediates the induction of alkaline phosphatase and mineralization by BMP-2 in pre-osteoblastic cells.
Introduction: Loss of function of LRP5 leads to osteoporosis (OPPG syndrome), and a specific point mutation in this same receptor results in high bone mass (HBM). Because LRP5 acts as a coreceptor for Wnt proteins, these findings suggest a crucial role for Wnt signaling in bone biology.
Materials and Methods: We have investigated the involvement of the Wnt/LRP5 cascade in osteoblast function by using the pluripotent mesenchymal cell lines C3H10T1/2, C2C12, and ST2 and the osteoblast cell line MC3T3-E1. Transfection experiments were carried out with a number of elements of the Wnt/LRP5 pathway. Measuring osteoblast and adipocyte differentiation markers addressed the effect of this cascade on osteoblast differentiation.
Results: In mesenchymal cells, only Wnt's capable of stabilizing β-catenin induced the expression of alkaline phosphatase (ALP). Wnt3a-mediated ALP induction was inhibited by overexpression of either Xdd1, dickkopf 1 (dkk1), or LRP5ΔC, indicating that canonical β-catenin signaling is responsible for this activity. The use of Noggin, a bone morphogenic protein (BMP) inhibitor, or cyclopamine, a Hedgehog inhibitor, revealed that the induction of ALP by Wnt is independent of these morphogenetic proteins and does not require de novo protein synthesis. In contrast, blocking Wnt/LRP5 signaling or protein synthesis inhibited the ability of both BMP-2 and Shh to induce ALP in mesenchymal cells. Moreover, BMP-2 enhanced Wnt1 and Wnt3a expression in our cells. In MC3T3-E1 cells, where endogenous ALP levels are maximal, antagonizing the Wnt/LRP5 pathway led to a decrease of ALP activity. In addition, overexpression of dkk1 reduced extracellular matrix mineralization in a BMP-2-dependent assay.
Conclusions: Our data strongly suggest that the capacity of BMP-2 and Shh to induce ALP relies on Wnt expression and the Wnt/LRP5 signaling cascade. Moreover the effects of BMP-2 on extracellular matrix mineralization by osteoblasts are mediated, at least in part, by the induction of a Wnt autocrine/paracrine loop. These results may help to explain the phenotype of OPPG patients and HBM.
Wnt proteins constitute a family of secreted cysteine-rich glycosylated proteins involved in a large variety of modeling and remodeling processes including cell polarity, cell differentiation, and cell migration.(1,2) Secreted Wnt's bind to and activate receptor complexes consisting of the frizzled family of G-protein-coupled receptors (GPCRs) and the low-density lipoprotein (LDL) receptor-related protein (LRP)-5 and LRP-6.(3) In the canonical Wnt/β-catenin pathway, activation of receptors results in the stabilization of β-catenin and its subsequent translocation into the nucleus where, in concert with transcription factors such as Tcf or Lef, it drives the transcription of target genes.(4) The ability of several Wnt's to stabilize β-catenin seems to be the basis for their proliferation and differentiation-dependent effects. Dysfunction of the Wnt/β-catenin pathway has been implicated in several diseases such as cancer and Alzheimer's disease.(5,6) More recently, it has been shown that loss of function of the Wnt coreceptor, LRP5, in both humans and mice leads to decreased bone formation postnatally,(7–9) and a point mutation in this same receptor results in a high bone mass trait.(10,11) These reports show the crucial role that Wnt/LRP5 signal transduction plays in bone accrual during growth and the importance of this signal for the establishment of peak bone mass. However, the exact mechanisms by which Wnt signaling members regulate bone formation still remain to be elucidated.
The bone-related phenotypic abnormalities found in human or mice lacking the LRP5 protein are largely caused by an osteoblast defect.(8,9) Although the transcription factor Runx2/Cbfa1 plays a pivotal role in osteoblast differentiation,(12,13) mice deficient in LRP5 normally express Cbfa1,(8) indicating that the mechanism by which LRP5 controls osteoblast function is Runx2/Cbfa1 independent. Interestingly, although several specific osteoblast genes are controlled by Runx2/Cbfa1, alkaline phosphatase (ALP) is not. Osteoblasts are the fully differentiated skeletal cells responsible for the production of bone matrix. Osteoblasts arise from mesenchymal stem cells that are pluripotential in nature and capable of giving rise to a number of committed and restricted cell lineages including osteoblast, chondroblast, adipoblast, fibroblast, and myoblast lines. We have recently demonstrated that a number of Wnt proteins are capable of inducing the osteoblast marker ALP in mesenchymal cells.(9) The same activity has been largely described for a number of morphogenic proteins including several bone morphogenic proteins (BMPs) and hedgehog (Hh) proteins. BMP-2 increases the level of β-catenin in the nucleus of pre-osteoblastic cells and induces the expression of Wnt15, 3a, 5b, and 7,(14) suggesting the existence of an interconnection between Wnt and BMP-2 signaling and/or their ability to induce osteoblast differentiation. Here we have determined the effect of the Wnt/LRP5 cascade on osteoblast function and the role of this signaling pathway in the induction of the osteoblast phenotype by BMP-2 and Shh. Our data clearly show that Wnt/LRP5 controls the expression of ALP through the canonical β-catenin cascade and that the capacity of BMP-2 and Shh to induce the expression of ALP in mesenchymal cells is dependent on the integrity of this pathway. Moreover we have demonstrated, by using the LRP5/6-selective antagonist dkk1, that Wnt/LRP5 signaling affects mineralization of osteoblast cells.
BMP-2 and N-terminal Shh production
BMP-2 was purified from Chinese hamster ovary (CHO) cells stably transfected with an expression vector encoding human BMP-2 as previously described.(15) The amino-terminal part of the murine Shh (corresponding to amino acids 25–198) was isolated by reverse transcriptase-polymerase chain reaction (RT-PCR) and confirmed by nucleotide sequencing. The isolated sequence was subcloned into the mammalian expression vector pABWN under control of CMV enhancer/chicken β-actin promoter (pShh). pShh was transfected into mouse fibroblast L-cells (LTK-P2 cells), and stably transfected cells were established. N-Shh was purified from cells culture medium as described by Spinella-Jaegle et al.(16)
Cell culture
C3H10T1/2 and ST2 (obtained from ATCC and MC3T3-E1), and MC3T3-E1 clone 4 (kindly provided by Dr R Franceschi) cell lines were cultured (5% CO2 at 37°C) in α-MEM supplemented with 10% heat-inactivated fetal calf serum. C2C12 cells (kindly provided by Dr G Karsenty) were maintained with 5% CO2 at 37°C in Dulbecco's modified Eagle's medium supplemented with 15% fetal calf serum. For treatment or transient transfection, cells were plated at 2 × 104/cm2, and 24 h later, culture medium was changed to 2% fetal calf serum. Treatment and transfections were carried out as indicated below.
Measurement of alkaline phosphatase activity, cell proliferation, and red-oil staining
Cells were treated for indicated time with BMP-2, Shh, or BMP-2/Shh. Alkaline phosphatase (ALP) activity was determined in cell lysates using an Alkaline Phosphatase Opt kit (Roche Molecular Biochemicals). Cell lysates were analyzed for protein content using a micro-BCA Assay kit (Pierce), and ALP activity was normalized for total protein concentration.
Cell proliferation measurements were performed by BrdU incorporation techniques using the cell proliferation ELISA Biotrak kit (Amersham). Cell proliferation was determined according to the manufacturer's protocol.
Determination of adipocytic cell formation was performed by staining intracellular triglyceride droplets. C3H10T1/2 cells were cultured as indicated above and then either left unstimulated or stimulated with an IDX cocktail (insulin at 1.6 μM, dexamethasone at 0.25 μM, and 3-isobutyl-1-methylxanthine (IBMX) at 0.5 μM). The accumulation of intracellular triglyceride droplets was visualized by staining with Oil Red.
Plasmids, cell transfection, and assay for luciferase activity
Murine Dkk1, β-catenin, disheveled, Wnt1, Wnt2, Wnt3a, Wnt4, Wnt5, Noggin, Gli3, Smad1, and LRP5 cells were isolated by RT-PCR and confirmed by nucleotide sequencing. Isolated sequences were HA-tagged subcloned into the expression plasmid pcDNA3, and expression was verified by Western blot using anti-influenza hemagglutinin (HA) antibodies. Expression of LRP5 constructs was also confirmed with an anti-LRP5/6 monoclonal antibody (BioVision). Dominant negative forms of disheveled (Xdd1) and Smad1 were generated as previously described.(16–18) Stable mutant β-catenin was generated as described by Morin et al.(19) The soluble LRP5ΔC form was generated as described by Gong et al.(9) CyclinD1 luciferase (cyclinD1/luc) reporter construct was generated as described by Leung et al.(20)
Cells were plated in 24-well plates as indicated above 24 h before transfection and transiently transfected with the indicated construct (1 μg total DNA) using DNA-lipid complex Fugene 6 (Boehringer Mannheim) according to the manufacturer's protocol. Sixteen hours after transfection, cells were washed and cultured in medium at 2% fetal calf serum for an additional 48 h under indicated conditions. Cells were assessed for ALP or luciferase activities. Controls were carried out by replacing constructs with empty vector.
When the luciferase reporter construct was used to assess transfection efficacy, 20 ng of pRL-TK (Promega) encoding a Renilla luciferase gene downstream of a minimal HSV-TK promoter was systematically added to the transfection mix. Luciferase assays were performed with the Dual Luciferase Assay Kit (Promega) according to the manufacturer's instructions. Ten microliters of cell lysate was assayed first for firefly luciferase and then for Renilla luciferase activity. Firefly luciferase activity was normalized to Renilla luciferase activity.
RNA extraction and RT-QPCR
Cells were treated as indicated above, and total RNA was isolated from cultured cells using an isolation kit from Quantum Appligene. Quantitative RT-PCR (RT-QPCR) was performed with TaqMan PCR reagent kits in the ABI PRISM 7700 Sequence detection system (Perkin-Elmer Applied Biosystems) as described by Gallea et al.(15) Primer/probe sets for the following target genes were designed using Primer-Express V1.0 software from Perkin-Elmer: ALP, osteocalcin (OC), collagen type I, Osf2/Cbfa1, PPARγ2, Wnt1, Wnt3a, frizzled1, and frizzled9. The sequences of the primer/probe sets are shown in Table 1. All RT-QPCR reactions were performed in duplicate, and the amplification signal from the target gene was normalized to a GAPDH signal in the same reaction. Data are presented as the relative mRNA variation in treated cells versus control cells.
Mineralization assay
For mineralization assays, MC3T3-E1 clone-4 cells were incubated in mineralization medium in 24-well culture plates at an initial density of 5 × 104 cells/cm2, as described by Wang et al. (α-MEM containing 10% or 2% FCS, and 50 μg/ml ascorbic acid).(21) Cells were either left untreated in the mineralization medium or were treated with BMP-2 (at indicated concentration) for 13 days. To induce extracellular matrix mineralization, 4 mM NaHPO4 was added 2 days before the end of the total 13-day culture period. The mineralized matrix was stained for calcium by Alizarin-red staining method as described by Stanford et al.(22) Briefly, after 13 days, cells were washed with PBS, followed by fixation in ice cold 70% ethanol for at least 1 h. Ethanol was removed and cells were rinsed with water and stained with 40 mM Alizarin-red (pH 4.2) for 10 minutes at room temperature. Stained cells were further processed by five rinses with water, followed by a 15-minute wash in PBS with rotation to reduce nonspecific Alizarin-red stain. Stained cultures were photographed, and Alizarin-Red fluorescence was measured on a multiplate reader (Vmax; Molecular Devices). Data are expressed as fluorescence arbitrary units.
Statistical analysis
All experimental data presented were obtained from two independent experiments, each in triplicate, and results are expressed as the mean ± SD. Comparisons between treatments were performed using Student's t-test. Values of p < 0.05 were considered statistically significant.
The canonical Wnt/β-catenin pathway induces the osteoblast marker ALP
One can distinguish between two types of Wnt proteins, β-catenin-dependent and β-catenin-independent Wnt's.(23) β-catenin-dependent Wnt's include Wnt1, Wnt2, Wnt3, and Wnt3a, whereas β-catenin-independent Wnt's include Wnt4, Wnt5a, Wnt5b, Wnt6, and Wnt7a (for more detail see http://www.stanford.edu/∼rnusse). We have tested the effect of both β-catenin-dependent and -independent Wnt's on three murine mesenchymal cell lines susceptible of acquiring the osteoblastic phenotype: C3H10T1/2, ST2, and C2C12. As depicted in Fig. 1, overexpression of Wnt3a resulted in a dramatic increase of ALP activity in C3H10T1/2, ST2, and C2C12 cells (Figs. 1A–1C). Overexpression of Wnt1 or Wnt2 resulted in an increase of ALP activity comparable with that induced by Wnt3a (data not shown). On the other hand, transient transfection of cells with the β-catenin-independent Wnt4 (data not shown) or Wnt5a (Fig. 1) did not show any significant effect on ALP expression in the cell lines used. Overexpression of β-catenin stable mutant(24) or treatment of cells with LiCl, which mimics Wnt signaling by inactivating GSK-3β thus increasing β-catenin cytoplasmic level,(25,26) significantly induced ALP expression in the three mesenchymal cell lines (data not shown).(9)
The involvement of the canonical Wnt cascade in the induction of ALP activity was further confirmed by experiments in which transfection with a dominant-negative form of Disheveled (Xdd1), an intracellular player of the Wnt signaling,(17,18) significantly reduced the Wnt-mediated induction of ALP activity in C3H10T1/2, ST2, and C2C12 cells (Figs. 1A–1C). Furthermore, transfection with the Wnt antagonist Dkk1(27–29) markedly decreased ALP induction by Wnt3a (Figs. 1A–1C). Dkk1 has been recently demonstrated to block the interaction of Wnt with LRP5 and LRP6.(30–32) Interestingly, overexpression of a dominant negative form of LRP5 (LRP5ΔC) abolished Wnt3a-mediated ALP induction (Figs. 1A–1C). The expression of Xdd1 and the expression and secretion of Dkk1 or LRP5ΔC constructs in our cell lines was monitored by Western blot technique in both cell lysate and cell supernatant (Fig. 1D and data not shown). These results show that Wnt3a-LRP5/6 interaction is necessary for ALP expression.
We have carried out RT-QPCR in our samples to investigate the effect of Wnt3a on the expression of a series of osteoblast markers. As expected, Wnt3a induces ALP gene expression in mesenchymal cells. However, Runx2/Cbfa1, OC, and collagen type I gene expression were unaffected by Wnt3a (data not shown).
Wnt3a inhibits adipocyte markers expression in C3H10T1/2 cells
The pluripotent mesenchymal C3H10T1/2 cells can differentiate into adipocytes,(16) and Wnt proteins are known to antagonize adipocyte differentiation. We have therefore investigated whether Wnt3a overexpression affects the expression of two adipocyte markers PPARγ2 and adipocyte fatty acid binding protein (aP2) in C3H10T1/2 cells. As shown in Fig. 2A, Wnt3a clearly inhibited the expression of PPARγ2 and aP2 expression as determined by RT-QPCR analysis. Accordingly, Wnt3a completely abolished the formation of lipidic vacuoles induced by IDX, an adipogenesis stimulator, as determined by Red-Oil staining (Fig. 2B).
Wnt3a induces morphological cell transformation and enhances proliferation of C3H10T1/2 cells
When C3H10T1/2 cells were transfected with a Wnt3a expressing construct, morphological transformation was observed within 24 h (Figs. 3A and 3B). The Wnt3a overexpressing cells showed increased cell density, became more refractile, and showed elongated morphology at high magnification. In contrast, C3H10T1/2 cells transfected with control plasmid remained morphologically unchanged (Figs. 3A and 3B).
We then investigated whether Wnt3a affects C3H10T1/2 cell proliferation. We first measured the ability of Wnt3a to modulate cyclinD1 transcriptional activity, an important factor in cell cycle and proliferation, by means of cyclinD1/luc reporter construct.(20) As shown in Fig. 3C, in C3H10T1/2 control cells, the basal level of cylinD1 transcriptional activity is high and Wnt3a overexpression enhances its activity by 2.5-fold. To confirm the ability of Wnt3a to enhance C3H10T1/2 cell proliferation, we have measured cell proliferation using a BrdU incorporation assay (Fig. 3D). In a time course (1–7 days) experiment, the measurement of BrdU incorporation shows that Wnt3a does not affect C3H10T1/2 cell proliferation at early stages but enhances cell proliferation after 6 days of culture.
BMP-2 induction of ALP is dependent on the canonical Wnt pathway
The morphogens BMP-2 and Shh have been shown to induce ALP activity in C3H10T1/2 cells,(15) and it is known that the BMP, Hh, and Wnt signaling pathways are interconnected.(33–35) We therefore investigated whether the ALP-inducing activity of Wnt3a is dependent on BMP or Hh signaling. For this purpose, we assessed the effect of Noggin, a well-characterized selective BMP inhibitor,(36) and cyclopamine, which inhibits Shh signaling,(37) on Wnt3a-induced ALP expression in C3H10T1/2 cells. As expected, transfection with Noggin abolished the induction of ALP by BMP-2 (Fig. 4A), and cyclopamine dose-dependently inhibited the ALP activity induced by Shh (Fig. 4B). However, neither Noggin nor cyclopamine affected the Wnt3a-mediated ALP activity (Fig. 4). In addition, the transcriptional activity of Smad1, which is involved in the intracellular BMP signaling,(38) was not modified by Wnt3a in the presence or absence of BMP-2 (data not shown). Finally, transfection of cells with Gli3, a Shh signaling inhibitor,(39) did not affect the ALP activity mediated by Wnt3a (Fig. 4C). Altogether, these data indicate that the induction of ALP by Wnt3a is independent of BMP and Hh signaling.
We then investigated whether Wnt signaling is required for BMPs or Hh to induce ALP expression. Interestingly, overexpression of the Wnt pathway inhibitors Xdd1, dkk1, and LRP5ΔC significantly reduced, and in the case of dkk1 almost abolished, the induction of ALP activity mediated by the two morphogens (Figs. 5A and 5B). These same constructs did not affect the capacity of retinoic acid to stimulate ALP activity in C3H10T1/2 cells, indicating that the induction of ALP by retinoic nuclear receptors is independent of the Wnt pathway. Thus, our results show that the integrity of the Wnt/LRP5 pathway is necessary for Shh and BMP-2 to induce ALP in mesenchymal pluripotent cells.
BMP-2 induces the expression of Wnt's
One possible explanation for the results described above is that Wnt-mediated signaling might directly control the expression of ALP. Pretreatment of C3H10T1/2 or C2C12 with the inhibitor of protein synthesis cycloheximide did not prevent the induction of ALP mRNA expression mediated by LiCl treatment (data not shown) and Wnt3a (Fig. 6). In contrast, the induction of ALP gene expression by BMP-2 was inhibited by cycloheximide (Fig. 6), indicating that de novo synthesis is necessary for this morphogen to induce ALP gene expression.
We then examined whether BMP-2 is capable of inducing the expression of Wnt's in C3H10T1/2 and ST2 cells. As shown in Fig. 7A, Wnt3a gene expression was increased by BMP-2 in both C3H10T1/2 and ST2 cells. Wnt1 gene expression was also found to be upregulated, but only in C3H10T1/2. In addition, we also investigated the expression of Wnt receptors, frizzleds (1–9), and LRP5 in response to BMP-2. LRP5 was expressed in the absence of BMP-2 and was not affected by this stimulus (data not shown). Our data show that only frizzled-1 (Fz1) and frizzled-9 (Fz9) expression was increased in the presence of BMP-2 (Fig. 7B). Altogether, our results strongly suggest that the effect of BMP-2 on ALP expression is mediated by a Wnt autocrine/paracrine loop.
ALP expression and mineralization of osteoblast-like cells MC3T3-E1 is controlled by Wnt signaling
To further investigate the role of Wnt/β-catenin signaling in osteoblast maturation, we examined the effects of Wnt3a in the osteoblastic-like cell line MC3T3-E1. As shown in Fig. 8A, Wnt3a overexpression did not modify the ALP activity already displayed by MC3T3-E1 cells. Overexpression of other Wnt proteins, such as Wnt1 and Wnt2, had no effect on ALP expression in these cells (data not shown). Although Wnt proteins had no effect on the already maximal ALP levels in MC3T3-E1 cells, Xdd1, LRP5ΔC, and Dkk1 significantly reduced the endogenous ALP activity displayed by these cells (Fig. 8A). We have therefore assessed the expression level of several Wnt mRNA in MC3T3-E1 cells. MC3T3-E1 cells were cultured for different time periods (2–5 days), total RNA was extracted, and the expression level of Wnt1, Wnt2, and Wnt3a was determined by RT-QPCR. The 2-day time-point was used as the reference point. ALP expression was found to be strongly and continuously increased over the time course (data not shown), as expected, with a cell committed toward the osteoblast phenotype. As shown in Fig. 8B, Wnt3a, but not Wnt1 or Wnt2, expression significantly increases over the MC3T3-E1 time course. This strongly supports the hypothesis that a Wnt/β-catenin autocrine/paracrine loop controls the expression of ALP in osteoblastic cells.
Although Wnt3a did not sensibly modify the mineralization capacity of MC3T3-E1 cells, we decided to evaluate the impact of inhibiting this signaling cascade on the mineralization of displayed by these cells in experimental conditions in which mineralization is dependent on exogenous BMP-2.(40) As shown in Fig. 8C, whereas no mineralization could be observed after 13 days of culture in the absence of BMP-2, 2.5–10 ng of BMP-2 clearly induced mineral nodules in these cells. Mineralization induced by BMP-2 could be quantified by mean Alizarin-red labeling and fluorescence measurement (Fig. 8D). Interestingly, transient overexpression of dkk1 in these cells significantly reduced the mineralizing capacity (Fig. 8D). Our results indicate that the Wnt/β-catenin pathway regulates osteoblast mineralization.
The recent discovery of the role of the Wnt coreceptor LRP5 in bone formation strongly suggests an involvement of the Wnt signaling pathway in skeletal biology.(8–11) Although different models show that Wnt signaling through the LRP5 receptor mainly affects osteoblast function, the precise molecular mechanisms remain to be elucidated. To gain further understanding of the control of osteoblast function by Wnt signaling, we investigated the effect of Wnt in both osteoblast commitment and differentiation by overexpressing different Wnt proteins in mesenchymal pluripotent cell lines capable of differentiating into the osteoblast lineage. Results presented herein clearly show that β-catenin-dependent Wnt's display the ability of inducing ALP expression in mesenchymal cells. The fact that LiCl was able to mimic Wnt ability to induce ALP expression and that dominant-negative disheveled inhibited Wnt ability to induce ALP expression clearly show that the capacity of Wnt's to induce ALP relies on the canonical β-catenin cascade. Moreover, the inhibition of Wnt effects, with either the LRP5/6 inhibitor dkk1 or a LRP5 dominant negative, highlight the crucial role that LRP5 plays in the induction of ALP expression by Wnt's.
Very interestingly, impairing Wnt/β-catenin signaling resulted in a significant reduction in the capacity of BMP-2 and Shh to induce ALP in these cells. These data strongly suggest that the Wnt/β-catenin pathway is required for the morphogenetic-mediated induction of ALP in mesenchymal cells. Further support for the requirement of Wnt signaling for the activity of BMP-2 activity comes from data showing that de novo protein synthesis is necessary for BMP-2 to induce ALP gene expression and that BMP-2 induces the expression of Wnt1 and Wnt3a in C3H10T1/2 cells. Our observation is consistent with the recent report that BMP-2 treatment upregulates expression of Wnt3a in high-density cultures of C3H10T1/2 cells.(41) Moreover, Hh signaling has also been shown to control the expression of Wnt proteins: Wnt5a is a target of Shh in hair follicle morphogenesis(42) and Gli1 (a crucial player in the mediation of Hedgehog signaling) has been shown to consistently induce a distinct set of Wnt genes in animal cap explants and skin tumors.(43) Very interestingly, microarray analysis of gene expression profile in different cell types demonstrated that Wnt proteins induce several genes that have been reported previously as BMP-2 target genes.(44,45) From these findings, it is expected that some activities of BMP-2 or Hh can be attributed to the effects of these proteins on Wnt gene expression. Our data suggest that BMP-2 and Shh do not directly regulate ALP expression but do so indirectly through the Wnt signal cascade. Figure 9 depicts a conceptual model for the proposed Wnt autocrine loop controlling ALP expression.
Although Wnt overexpression did not affect the already high endogenous ALP activity displayed by the osteoblastic cells MC3T3-E1, RT-QPCR clearly shows that these cells endogenously express Wnt3a. In addition, disruption of Wnt signaling by overexpressing Dkk1 significantly reduced both ALP activity levels and the capacity of mineralization of these cells, showing the existence of an autocrine/paracrine loop in osteoblasts. Consistently, in vitro mineralization of the extracellular matrix by LRP5−/− mouse-derived osteoblasts was delayed compared with osteoblasts from wild-type animals.(8) This is due either to a functional defect in osteoblasts deprived from LRP5 or an abnormal proliferation rate in knockout animal-derived cells.(8) Although Wnt3a significantly enhances mesenchymal cell proliferation, this effect occurs very late compared with BMP-2. Our data concerning the control of ALP by Wnt signaling suggest that the absence of LRP5 affects the osteoblast function by reducing ALP activity, and subsequently, the mineralization capacity of these cells. It is important to point out that it has been recently reported that transgenic mice expressing the human high bone mass gene display an enhanced ALP staining in osteoblasts.(46) Therefore, the control of ALP expression by Wnt signaling in both pluripotent mesenchymal cells and osteoblasts might partially explain the skeletal phenotypes occurring in human or mice displaying LRP5 mutations.
In contrast with ALP, other osteoblast markers such as Runx2/Cbfa1 and osteocalcin were not affected by Wnt overexpression. This is consistent with the normal expression of these two osteoblast markers reported in LRP5−/− mice(8) and strongly suggests that Wnt signaling controls osteoblast function in a Runx2/Cbfa1-independent manner. Although elevated levels of serum osteocalcin have been found in a kindred affected with high bone mass disorder,(11) no osteocalcin data have been reported from transgenic mice overexpressing the human LRP5 HBM mutation.(46) It would be interesting to measure osteocalcin levels in these transgenic mice but also in other human kindred displaying either the HBM trait or OPPG phenotype. In any case, our results suggest that Wnt signaling regulates osteoblast commitment and function without interfering with Runx2/Cbfa1 or osteocalcin expression, and it constitutes a Runx2/Cbfa1-independent pathway crucial for osteoblast maturation. It has been previously reported that Runx2 overexpression induces ALP gene expression in C3H10T1/2 cells.(47) In the case of C2C12 cells, Lee et al.(48) found a slight increase in ALP activity in cells overexpressing Runx2, whereas Zhang et al.(49) did not observe detectable levels of ALP activity in C2C12 stably expressing Runx2. In our hands, Runx2 overexpression did not induce any detectable ALP activity in these two cell lines. One possibility is that cell lines used in other studies could spontaneously secrete some BMP proteins. In this case, Runx2 overexpression can enhance ALP activity. Actually a cooperation between Smads and Runx2 has been suggested by Zhang et al.(49) In any case, our experimental data using dkk1 clearly show that the integrity of Wnt/LRP signaling cascade is necessary for induction of ALP by BMP-2.
We also present evidence here that Wnt3a can compromise the adipocytic commitment of the pluripotent mesenchymal cell line C3H10T1/2. Importantly, Wnt3a dramatically reduced the levels of PPARγ2 and aP2, two genes playing a central role in adipogenesis. Although it is well established that Wnt signaling inhibits adipogenesis,(50) this is the first report to our knowledge showing the inhibition of adipogenesis in cells displaying an osteoblastic potential. Whereas BMP-2 has been shown to induce differentiation of C3H10T1/2 cells into adipocyte and osteoblasts,(21) Hh(16) and Wnt3a (this study) display opposite actions in adipogenesis and osteogenesis. Our hypothesis is that signaling cascades triggered by these proteins lead, respectively, to the repression of adipocyte master genes and activation of genes playing crucial roles in osteoblast differentiation such as ALP. Osteoblasts and adipocytes differentiate from the same progenitor cells, and pathways that regulate osteoblastic differentiation versus adipogenesis are of high importance. Total marrow fat increases with age, and there is an inverse relationship between marrow adipocytes and osteoblasts with aging.(51,52) The number of mesenchymal stem cells with osteogenic potential decreases early during aging in humans and may be responsible for the age-related reduction in osteoblast number.(53) Importantly, it has been demonstrated that cells cultured from human trabecular bone are not only osteogenic but are also able to undergo adipocytic differentiation under defined culture conditions.(54) It is therefore attractive to speculate that Wnt signaling constitutes one of the mechanisms to control the relative commitment of cells into either the osteoblast or the adipocytic lineage. Further investigation is required to determine the possibility of a switch from adipocyte to osteoblast commitment in a given cell. Interestingly, in the ΔFosB transgenic model (increased bone formation and inhibited adipogenesis), osteosclerosis has been shown to be independent of decreased adipogenesis.(55)
Few anabolic agents have been developed as therapeutics for skeletal disorders. Because of the crucial role of the Wnt signaling pathway for osteoblast activity, this cascade might constitute an important new target for anabolic drug development. To discover or design drugs selectively affecting this pathway, we need to elucidate the mechanisms by which the Wnt cascade affects pre-osteoblasts and osteoblast function. In this study, we contribute to the understanding of these molecular effects by showing (1) the crucial role that Wnt signaling plays in both the acquisition of ALP gene expression by mesenchymal pluripotent cell lines and the control of ALP expression and mineralization of osteoblasts, (2) the importance of LRP5/Wnt signaling in the morphogenetic protein-mediated induction of ALP in mesenchymal cells, with the consecutive impact in the commitment of these pluripotent cells into the osteoblastic lineage, and (3) the effect of Wnt cascade in inhibiting adipogenesis in mesenchymal pluripotent cells.
We would like to thank Dr Chantale Bardelay and M Berbard Doucet for help in adipocytic cell staining. We are also indebted to Veronique Stiot for technical help.
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High solids hydrophobically modified alkali-soluble emulsion (HASE). Used as a premium rheology modifier to provide maximum high-shear viscosity for improved film build in interior and exterior waterborne coatings formulations. Offers ease of incorporation and rapid viscosity. Possesses low odor and zero VOC. Provides very good flow, leveling and spatter resistance, high gloss development and microbial and mildew resistance. Can be used in combination with other HASE and cellulosic thickeners for optimum enhanced high-shear viscosity in medium and high PVC vinyl acrylic coatings. Can also be used as the sole thickener in small particle size acrylic or styrene acrylic formulations for maximum film build and gloss development.
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US Condemns Aleppo Barrel Bomb Raids
US Secretary of State John Kerry has condemned the use of barrel bombs in the Syrian city of Aleppo by forces loyal to President Bashar al-Assad.
Kerry said it was the "latest barbaric act of the Syrian regime".
The comments came after Syrian army helicopters reportedly dropped more barrel bombs on Aleppo on Tuesday, killing at least eight people.
"Each and every day that the barrel-bombing of Aleppo continues, the Assad regime reminds the world of its true colors," Kerry said.
"It is the latest barbaric act of a regime that has committed organised, wholesale torture, used chemical weapons, and is starving whole communities by blocking delivery of food to Syrian civilians in urgent need," he said.
"Given this horrific legacy, the Syrian people would never accept as legitimate a government including Assad."
The US and its allies have been supporting the opposition's call for a transitional government that does not include President Assad. Syria's ally, Russia, has been resisting the move.
"While the opposition and the international community are focused on ending the war... the regime is single-mindedly focused on inflicting further destruction to strengthen its hand on the battlefield and undermining hopes for the success of the Geneva II process," Mr Kerry added.
Aleppo has been divided since rebels captured large swathes in 2012. Much of the Old City has been levelled by fighting.
Talks between the Syrian opposition and the government took place last week in Geneva but ended without result.
A further round is planned for next week.
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- » Helicopter with Thirteen People on Board Crashes off Norway
- » Austria Adopts New Tougher Asylum Law


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September 1, 2015
Hain Celestial: elected Raymond W. Kelly to its board of directors. Kelly, who served as the police commissioner of New York City for 14 years, has been president of risk management services for Cushman and Wakefield, Inc. since March 2014. Cushman a…
September 1, 2015
ISSA: the worldwide cleaning industry association, appointed John Barrett executive director, effective Sept. 1, 2015. Barrett replaces John Garfinkel, who will work in a consultative capacity through 2016 to facilitate a smooth transition. Un…
September 1, 2015
CBX: appointed veteran account director and business developer Jodi Sutherland as vice president, engagement group director. In her new position, Sutherland will partner with peer leaders at CBX to market the firm to key audiences and develop busines…
September 1, 2015
Bioelements: welcomed Jessica Finlay as its new content marketing writer and coordinator. She joins the marketing team at Bioelements with vast experience in digital publishing and marketing. In her new role, Finlay will apply her expertise in web co…
September 1, 2015
United Mineral and Chemical Corporation (UMC): appointed Michael Sansonetti Jr. as its new president. This came after the announcement that UMC's long-time president, A. Nurhan Becidyan, would retire. Sansonetti, who is currently in charge of…
September 1, 2015
Markwins: appointed Jeff Rogers as president of Physicians Formula Group. He succeeds Ingrid Jackel, who left the company to serve as CEO of Yes To. Rogers will continue to work closely with Markwins founder and CEO Eric Chen to further elevate the…
September 1, 2015
Shiseido Co., Ltd.: named Marc Rey president and CEO of Shiseido Americas. In addition to this role, Rey will also have global brand responsibility for Nars Cosmetics and Bare Escentuals. He previously served as president of Coty USA and regional sen…
September 1, 2015
Hutchinson has more than 20 years of experience in the natural products industry and will oversee sales and marketing for the brand, which is available in 2,500 retail stores throughout North America and Asia. Jantzen, who has spent the past three y…
September 1, 2015
Procter & Gamble (P&G): named David S. Taylor, group president of global beauty, grooming and health care, as successor to A.G. Lafley as president and CEO, effective Nov. 1, 2015. Taylor has also been appointed to the company's b…
August 26, 2015
Adds two new employees in sales and marketing.
August 26, 2015
Klinger, formerly of Merck, to lead finances in 2016.
August 19, 2015
Barrett to replace Garfinkel after 25 years of leadership.
August 18, 2015
FMCG company adds Raymond Kelly.
August 14, 2015
Manuel Gonzalez is chief commercial officer.
August 12, 2015
Hutchinson, Janzen and Flores are on the rise.
Knowlton Development Corporation [KDC] is North America's leading personal care contract manufacturer and custom formulator.
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My husband and I would like to take my son and daughter (23 and 18) to Puerto Rico in early January (only time we can coordinate such an event). We'd like to rent a two bedroom, two bath place, hopefully near beaches, but also near to life, culture and fun so that our kids won't feel like they are isolated in a retirement community. Frankly, I am overwhelmed by the choices on sites such as VRBO etc., and I would love some help narrowing down the options. Any suggestions?
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National News
The auditor-general has issued a highly critical report on the New Zealand Defence Force (NZDF) cuts, saying they will fail to meet savings targets and have led to a drop in morale and capability.
The Government told the Defence Force in 2010 to reduce costs so money could be redistributed within the military. The civilianisation project was one of several the defence force initiated in response.
But Auditor-General Lyn Provost said today it had achieved only "limited success".
"It will not achieve the NZDF's target of converting 1400 military positions and saving $20.5 milion a year by 2014/15.
"Instead, 600 military positions will be converted and we estimate savings of $14.2m a year by 2014/15. In addition a drop in morale and increase in staff attrition has led to reduced NZDF capability."
Her report was critical on numerous fronts - including the fact it told the Government it would convert 1400 military positions into civilian jobs without knowing how many military positions it would need from 2015.
After an investigation NZDF discovered that while some ranks and trades had surplus military staff, it was going to need more military staff overall.
"NZDF should have found out how many and what kind of military staff it would need before telling the Government that it would convert 1400 military positions into civilian positions," Provost said.
Despite the above, NZDF was standing by its commitment to the Government that it would achieve $350-$400m in total savings. But in the meantime it had lost far more military staff than intended.
"The loss of so many military staff (which can be attributed in part to the civilianisation project), has made it more difficult for NZDF to do its job," Provost said.
"Converting 1400 military positions into civilian positions would always be difficult. Discharging military staff has to be carried out with great care to avoid damaging the bonds of camaraderie, integrity and commitment that are part of NZDF culture.
"Instead, NZDF chose a course that led to a drop in morale and an increase in attrition resulting in reduced capability. NZDF now needs to recover from the damage caused by the civilianisation project."
Provost was critical of the speed at which NZDF went about the savings and said it did not fully consider the project's potential impact on staff, or address the significant risks of the process.
However, NZDF appeared to have learnt from its mistakes.
"My staff saw much evidence in reviews and in briefings to Cabinet and the Minister of Defence that NZDF recognises that it made mistakes... NZDF has decided that further conversion of military positions to civilian positions will, in general, take place gradually, as staff leave particular positions. NZDF has a focus on rebuilding morale and restoring mutual trust with military staff.''
New Zealand Defence Force Lieutenant General Rhys Jones said the Auditor-General's report repeated messaged they already knew about.
"The observations and conclusions made by the Auditor General mirror in large part what our own people had been telling us. When combined with some of the conditions of service changes being considered at the same time, many within the Defence Force felt that we had broken a social contract with them."
While the plan had been the civilianisation 1400 positions, only 303 military staff were discharged - 87 into civilian jobs and 218 were made redundant.
The impact on morale was the reason the programme was changed after the first phase, Jones said.
"To date the Defence Force's reforms have delivered nearly $160 million, which has been a massive undertaking. Civilianisation through attrition has continued, and we believe the financial targets we set ourselves when we started, will be met."
The civilianisation programme had a "detrimental affect" on the perception of the organisations culture, he said.
Efforts were now being made to rebuild morale, including with a pay increase in 2012.
"However, the reality of militaries throughout the world, such as the United Kingdom and the United States, is that down-sizing is a fact of life. Our Defence Force hadn't had to deal with that for a while, so it came as a shock."
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1. Pensioner, pensionary one who receives a pension. (m & f)
past participle
2. Past participle of PENSIONAR.
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10 Payers With the Lowest Denial Rates
Cloud-based software services provider athenahealth has released its 2014 PayerView Report, which leverages the company's data to provide insight into provider-payer relationships and ranks healthcare payers based on various metrics.
Read the full article on Becker's Hospital Review.
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- Tenet officials unfazed by UnitedHealth's ACA departure — 3 points
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- Surgery Partners' short interest falls 4.01%: 5 notes


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Hunter Hawk 4 Wood Burning Stove
Price £486.00Inc. VAT
Hunter Hawk 4 Stove with traditional door
|View all Hunter Stoves
Visit Hunter Website
Ask us about this product
The air-wash is achieved by air being drawn into the top of the stove and heated in a large air chamber before being directed down behind the large glass door creating a screen between the fire and the glass. Smoke or combustion particles do not come into contact with the glass, leaving it clean to see through.
For efficient woodburning the hot airwash is directed down the stove door and into the heart of the wood fire, this ensures complete combustion which helps to keep the chimney clean - the optimum conditions for burning wood.
By reducing the air inflow to an absolute minimum, the fire will gently smolder and remain alight throughout the night. When required, the fire will come to life by simply increasing the air flow into the stove.
This means the stove can burn wood and smokeless fuels in a Smoke Controlled Area
The Hunter Hawk stove has a number of options and optional extras :
* Traditional (4) or Contemporary (4D) door design.
* Optional slide on Low Canopy for an alternative look. This will add 145mm to the height of the stove.
* A flue damper - To help reduce excessive chimney draw and aid overnight burning.
* A solid brass door knob.
* Matt black finish with optional dark green or metallic blue.
* Defra Approved
PLEASE NOTE : The flue collar provided by Hunter is now a plain collar rather than the decorative collar as shown in many of the product images.
NB: Flue pipe is not provided by the manufacturer with most stoves. Exceptions to this are flueless gas stoves and electric stoves which do not require flue pipe. The inclusion of flue pipe with balanced flue gas stoves varies and should be confirmed dependant on the model ordered. You can order flue pipe from the 'Flue & Chimney' menu option at the top of the page.
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Thursday, 5 May 2016
Last updated 13 hours ago
Jul 21 2009 | 7:04am ET
A former top executive at defunct hedge fund shop D.B. Zwirn & Co. says he got a bad rap and a bum I.O.U. from firm namesake Daniel Zwirn.
Perry Gruss filed suit against Zwirn, who turned over what was left in his hedge funds to Fortress Investment Group in May, accusing him of defamation and breach of contract. The suit alleges that Zwirn made Gruss a "scapegoat" for the purchase of a private jet with investor money without their knowledge, and says that Zwirn's statements have kept Gruss from finding a new job since his "forced exit" from D.B. Zwirn three years ago.
According to the lawsuit, Zwirn told Gruss that he had to go so that the firm's namesake could remain "pearly white" and "bullet-proof."
"With Gruss out of the way, investors were told a series of false and misleading statements which sought to blame Gruss for any conceivable control issues at the company, and to shift the blame to Gruss without implicating Zwirn," the suit, filed in Manhattan federal court, alleges.
Gruss also claims that Zwirn failed to make good on payments "in connection with his forced exit" from the New York-based firm, which shuttered its largest hedge funds last year.
Gruss is seeking $9.6 million in damages.
May 3 2016 | 6:11pm ET
The cadre of 25 research analysts at Murano Systems speaks with more than 150 investors...
Apr 6 2016 | 8:31pm ET
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Apr 18 2016 | 5:51pm ET
The recent decision by the New York City Employment Retirement System to exit its...


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Cloud Infographic: Google Drive Vs The Competition
There are hundreds of cloud services available today, a fact which does not make it any easier for users to select a cloud storage provider that fits their needs.
Google is no longer the only kid on the cloud computing block. In fact, Apple, Microsoft, Amazon and even private parties are entering into the cloud storage industry, and rather aggressively. The race is on for these cloud storage services to bring in the most customers and attract the most attention, but who is the best of the best? Answering this question is not as simple as one might hope. In fact, there's a lot more to selecting the best cloud storage company than meets the eye. This is because cloud storage is customized to users based on their distinct needs, which means while a certain cloud storage may be perfect for one individual, it might not work well for another.
- The Internet of Everything: Why The IoT Will Take Over Every Industry - April 29, 2016
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- Mondo Report: The Top 5 Cloud Positions/Skills In Highest Demand - April 5, 2016


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Dentons prepares to seal transatlantic merger with Sonnenschein
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Uncle Pelican 26 May 2010 at 22:41
The noise that the newly created firm will hope to make on the world stage will surely be somewhat muted by a zero presence in that engine of global growth – Asia-Pacific – especially given the mega energy deals going on there recently. Would the new firm perhaps seek a third alliance in the region to give itself truly global capability?
Anonymous 27 May 2010 at 08:33
Having trained with DWS, I have a great deal of respect for the place. I just can't see what the merger really has to offer.
There appears to be very little synergy between the practice groups and their metrics are very different.
SNR is a top drawer insurance litigator, but DWS (when I was a trainee) had only one partner practicing this discipline in London.
Both firms market a focus on IPMT, but as most people at DWS would tell you, IPMT contributes a very small % of turnover.
DWS strengths in London are mid-market finance (save for the excellent Wynne, Felberbaum, Andrews, Paine and Rice), real estate and a clutch of energy/project lawyers in London, Paris and Dubai. Frankly I can't see how SNR's St Louis and Walnut Creek offices really advance these practice areas.
When Dentons and Wilde Sapte merged, a lot of top partners left because the costs were going to cripple them. I hope the same doesn't happen again.
Anonymous 27 May 2010 at 11:04
@ Dave Harris wrote Dentons could have held out for better, IMO. Perhaps Howard Morris wanted a merger where he could be in the driving seat?
Denton's are merging with a firm with a PEP more than twice as high, a far higher RPL and around twice as big in total revenues.
This is actually a wonderful merger for Dentons.
Strategically is also great, creating a firm with a sizeable footprint in both Europe and the U.S. and the resources to expand in both and also look towards Asia.
Wilde Sapte 27 May 2010 at 11:25
@Anonymous 8.33am, I think you may have gotten the wrong end of the stick. It's not about the practice fits but what you do with them. If their strategy is to target the Middle East and Asia then a larger platform makes sense. In any event, there is a good practice fit on the energy side.
Dave Cameron 27 May 2010 at 12:50
That video is HILARIOUS!!!!!! Do you think they did it all in one take? It's amazing the way they read each other's minds, answering questions without any questions being asked.
It's definitely worth watching right to the end for Howard Morris's "I'm so excited by this merger" finale. I'd hate to see him bored by something……
Joe C 27 May 2010 at 16:45
Please do not foget that SNR New York is the former high powered New York finance practice of Thacher Profit and Wood, which is a natural fit to the DWS London office. The TPW team has only recently joined so there is a lot of growth for that. SNR is as much a NY firm as it is a Chicago firm. Along with the DWS growth markets this will make a powerful combination in the long term. Give it a couple of years I think these guys are going to be heard.
Qasim11876 29 May 2010 at 15:25
We might presume that they will have an eye to move into the Asian market in the near future.


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Jul 10, 2015
FaithAction International House, a CLINIC affiliate led by Reverend David Fraccaro, is making great strides in promoting and encouraging immigrant integration within the city of Greensboro, North Carolina.
Jul 7, 2015
Jul 2, 2015
Featured Resources
The Catholic Legal Immigration Network, Inc., (CLINIC) has produced an information sheet to help people understand the recent enforcement actions by Immigration and Customs Enforcement (ICE) to take into custody and deport families. Although the Jan. 2-3 actions took place in a handful of states, they have led to an environment of confusion and...
Jan 8, 2016


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