We describe a method for subcellular fractionation of mouse skeletal muscle, myoblast and myotubes to obtain relatively pure fractions of nuclear, cytosolic and mitochondrial compartments. Fractionation allows the analysis of a protein of interest (or other cellular component) based on its subcellular compartmental distribution and can also generate molecular information about the state of a cell and/or tissue and how the distribution of a protein may differ between different cellular compartments, tissues or cell types, in response to treatments or ageing.
The described method was specifically developed for skeletal muscle and proliferating/differentiated muscle cells. The purity of the different fractions, representing the cytoplasmic, mitochondrial and nuclear subcellular compartments was validated by western blot analysis of "house-keeper" marker proteins specific for each cellular compartment.
This low cost method allowed the mitochondrial, cytoplasmic and nuclear subcellular compartments from the same starting muscle samples to be rapidly and simultaneously isolated with good purity and without the use of an ultracentrifuge. This method permits samples to be frozen at −80°C for future analysis and/or additional processing at a later date.
Isolation of nuclear, cytosolic and mitochondrial fractions of reasonable purity from mammalian tissues and cells has generated great interest as it has the advantage of allowing different cellular proteins and organelles to be studied and characterised. Subcellular fractionation is universally used for various cell types and tissues for sample preparation and prior to subsequent ~ omics analysis [1–5]. Generic fractionation protocols exist that can purify specific subcellular compartments and organelles, but in general they are not tailored for use with skeletal muscle and may require large amounts of starting material, time, or special reagents whilst potentially yielding fewer fractions from the same starting sample etc. [3, 5–9]. The protocol described has been optimized for use with primary skeletal muscle tissue (e.g. mouse anterior tibialis (AT) muscle) and both proliferating and differentiated C2C12 cells to isolate subcellular fractions of nuclei, cytosol, and mitochondria from a single starting sample, thereby reducing the quantity of starting material, cost and total time needed for sample preparation.
The protocol works well for skeletal muscle tissue and cells and could be used as a starting point for the fractionation of other non-muscle samples although changes to buffer volumes; homogenization duration/intensity etc. may be required. The purity of the fractions obtained was assessed by immunoblotting for specific protein markers: histone H3 (nuclei), glyceraldehyde 3-phosphate dehydrogenase (GAPDH, cytosol), and cytochrome oxidase IV (CoxIV, mitochondria).
The C2C12 mouse skeletal myoblast cell line was obtained from the American Type Culture Collection (CRL-1772). C2C12 myoblasts were maintained in DMEM (Sigma Aldrich, Poole, UK) supplemented with 1% L-glutamine (Lonza, Cologne, Germany), 10% FBS (Biosera, Sussex, UK) and 1% penicillin and streptomycin (Sigma) under an atmosphere of 5% CO2 in humidified air at 37°C. To induce myogenic differentiation, the growth medium was changed to differentiation medium (DMEM supplemented with 2% horse serum (Sigma) and 1% antibiotics) after myoblasts had reached ≈ 90% confluence in a T75 cm2 flask. Myoblast cells were either harvested at 90% confluence or allowed to mature to myotubes for 7 days and then harvested (see below).
Adult mice (C57BL/6) were euthanized by overdose with anesthetic (ketamine hydrochloride and medatomidine hydrochloride) administered by intraperitoneal injection. Anterior tibialis (AT) muscles, approximately 50 mg wet weight, were rapidly removed and used fresh to prepare fractions. Experiments were performed in accordance with UK Home Office Guidelines under the UK Animals (Scientific Procedures) Act 1986 and received ethical approval from the University of Liverpool Animal Welfare Committee.
Fresh AT tissue and scraped cells were washed with cold PBS, cells were pelleted by centrifugation at 200 g for 7 minutes whereas tissues were placed in a pre-chilled glass Petri dish and minced on ice using sharp scissors. All samples were resuspended in 300-500 μl of STM buffer comprising 250 mM sucrose, 50 mM Tris–HCl pH 7.4, 5 mM MgCl2, protease and phosphatase inhibitor cocktails (all chemicals were from Sigma-Aldrich, Poole, UK unless stated otherwise) and homogenized for 1 minute on ice using a tight-fitting Teflon pestle attached to a Potter S homogeniser (Sartorius Stedium, Goettingen, Germany) set to 600–1,000 rpm. The homogenate was then inspected, if intact tissue was still evident the homogenisation was repeated. The homogenate was decanted into a centrifuge tube and maintained on ice for 30 minutes, vortexed at maximum speed for 15 seconds and then centrifuged at 800 g for 15 minutes. The pellet was labelled as P0 and kept on ice, the supernatant was labelled as S0 and used for subsequent isolation of mitochondrial and cytosolic (Figure 1) fractions.
Schematic representation of the fractionation protocol. The developed protocol provides three subcellular fractions of cytoplasm, nuclei and mitochondria from a muscle sample. (− − −) Dotted arrow shows an optional step.
The pellet P0 (containing nuclei and debris) was resuspended in 300-500 μl STM buffer, vortexed at maximum speed for 15 seconds and then centrifuged at 500 g for 15 minutes. Following the above step, the nuclear pellet was labelled as P1 and kept on ice, the supernatant S1 (cell debris) was discarded. The purity of the nuclei within fraction P1 can be quickly determined by microscopic inspection by diluting an aliquot of the fraction in a trypan blue solution on a haemocytometer. If the P1 fraction contained excess cell debris the above step was repeated once.
To increase the P1 fraction purity further it was washed in STM buffer (300-500 μl), vortexed at maximum speed for 15 seconds and then centrifuged at 1,000 g for 15 minutes. The washed pellet was labelled as P5 (S5 was discarded) and resuspended in 200-500 μl NET buffer (comprising: 20 mM HEPES pH 7.9, 1.5 mM MgCl2, 0.5 M NaCl, 0.2 mM EDTA, 20% glycerol, 1% Triton-X-100, protease and phosphatase inhibitors) using a pipette to triturate until homogeneous. Pellet P5 was vortexed at maximum speed for 15 seconds and incubated on ice for 30 minutes, this fraction contained the nuclei. The nuclei were lysed with 10–20 passages through an 18-gauge needle and/or sonicated (using a Soniprep 150, MSE, London, UK) at high setting for 10–15 seconds with 30 second pauses whilst being kept on ice throughout. The lysate was centrifuged at 9,000 g for 30 minutes (at 4°C), the resultant supernatant (S6) was the final "nuclear fraction" (Figure 1).
Cytosolic and mitochondrial fractions were extracted from S0 by centrifugation at 800 g for 10 minutes. The supernatant S2 was saved and the pellet (P2) was discarded, though to improve the nuclear yield the pellet P2 can be combined with fraction P0 (optional step). S2 was then centrifuged at 11,000 g for 10 minutes and the supernatant S3 (containing cytosol and microsomal fraction) was precipitated in cold 100% acetone at −20°C for at least 1 hour followed by centrifugation at 12,000 g for 5 minutes, the pellet (P7) was then resuspended in 100-300 μl STM buffer and labelled as "cytosolic fraction" (Figure 1) that likely included some microsomal content. The pellet P3 was again resuspend in 100-200 μl STM buffer and centrifuged at 11,000 g for 10 minutes. Once centrifuged, supernatant S4 was discarded, the mitochondrial pellet (P4) was resuspended in 50-100 μl SOL buffer (comprising: 50 mM Tris HCl pH 6.8, 1 mM EDTA, 0.5% Triton-X-100, protease and phosphatase inhibitors) by sonication on ice at high setting for 5–10 seconds with 30 second pauses and labelled as "mitochondrial fraction". All buffers and centrifugation steps were modified from Cox and Emili  and Psarra et al. .
Additionally, a commercial cell fractionation kit designed to yield near pure nuclei and cytoplasmic fractions from the same starting sample of cells and soft tissues was used (Thermo NE-PER nuclear and cytoplasmic extraction kit, Pierce-Thermo, Northumberland, UK, to obtain a mitochondrial fraction required the use of an additional kit) as directed by the manufacturer. This enabled a comparison of the efficacy of the commercial kit with the method described here when both methods were used to fractionate an identical starting sample (myoblast cells from a T75 cm2 flask, approximately 2×106 cells).
The protein content of each compartment was determined using BCA protein assay (Sigma).
Validation of the purity of the subcellular fractions derived from the same starting sample was determined by examining "house-keeper" (HK) protein markers by standard SDS-PAGE analysis. Protein samples (15–20 μg/fraction) were separated by a 12% SDS/PAGE and transferred onto a nitrocellulose membrane (Sigma) and probed using monoclonal antibodies for Histone H3 (Cell Signalling, Hertfordshire, UK, 1: 2,000), GAPDH (Abcam, Cambridge, UK, 1: 5,000), and Cox IV (Abcam, Cambridge, UK, 1: 2,000) for nuclei, cytosolic and mitochondrial HK fractions respectively. Proteins were then visualized after applying specific secondary HRP-conjugated antibodies and exposure to a supersignal west dura substrate (Pierce-Thermo, Northumberland, UK) by use of a ChemiDocTM XRS (Bio-Rad, Hertfordshire, UK) see Figure 2 (representative example of at least three replicates per sample). Secondary antibody controls were also investigated and no non-specific binding was apparent (data not shown). To assess the purity of each fraction, bands relative to sub-compartment protein marker were quantified by ImageJ software .
Example western blots from three starting samples. In the top panel; (A) myoblast sample (20 μg protein was added to each lane) fractionated into nuclei, cytoplasmic and mitochondrial samples. Each fraction was run side-by-side on the same blot and then probed separately against each of the three primary antibodies, Histone-H3, GAPDH and CoxIV, to validate fraction purity. This was repeated for; (B) myotubes (20 μg protein per lane) and (C) AT tissue (15 μg protein per lane). In the bottom panel; marker protein fraction expression was measured as OD per resultant band area and was expressed in arbitrary units. The histogram data are representative of the mean (± SEM) of five separate experiments.
After fractionation and extraction of proteins from the C2C12 cell cultures and AT muscles using the protocol described in Figure 1, we report the mean fraction yield values obtained from replicate experiments (n = 5) (Table 1). Figure 2A-C show the nuclear, cytoplasmic and mitochondrial fractions obtained from a single starting sample from each of the different tissue/cell types. The fractions were examined by western blotting analysis using antibodies directed against specific HK markers and revealed single dense bands for histone H3 at approximately 17 kDa in the nuclear fraction, a band representing GAPDH at approximately 37 kDa in the cytosolic fraction, and a single band at approximately 17 kDa for CoxIV in the mitochondrial fraction (Figure 2A-C) in each sample.
Typical protein yield (μg) obtained after sample fractionation using the indicated volume of lysis buffer for myoblasts, myotubes (both cultured in 75 cm2 flask, ≈2×106 cells) and AT muscle (all n = 5, mean ± SEM). The protein yields (determined by BCA assay as total volume of a fraction × protein concentration in the fraction) are representative and dependent upon the volume of buffer utilized to resuspend each fraction.
These analyses showed little contamination between compartment fractions indicating that the purities of the nuclear, cytosolic, and mitochondrial fractions were relatively high (Table 2). The HK band intensity was high arguing that enrichment of proteins was achieved during processing (Table 1). The nuclear and mitochondrial fractions showed the highest band density indicating good yields and the lack of equivalent molecular weight bands in other fractions argued relatively little cross-talk and indicated reasonable purity, with the myotubes being slightly more variable depending on the fraction (feint additional non-specific bands were evident in the cytosolic fraction from myotubes). Furthermore, analysis of the distribution of the sub-compartment fraction marker proteins (Table 2) showed the minimum percentage of purity was ≈ 85.6% (Cox IV mitochondrial marker in myoblasts) and therefore argued less than 15% was from other sources.
The percentage shown in the table is obtained by the ratio between OD values of Histone H3, GAPDH and CoxIV (all n = 5, mean ± SEM) in an individual target compartment divided by the sum of the OD values present in all compartments in each sample.
Comparison of the method described with a commercially available fractionation kit showed both approaches generated clean and abundant cytoplasmic fractions (Figure 3A), although the yield using the kit was smaller. The nuclear fraction generated by the kit showed both reduced yield and compromised purity (Figure 3B). The kit was not designed to generate a mitochondrial fraction and so no equivalent comparison was undertaken.
Comparison of fractions generated from a muscle source by a kit versus the described method. Example western blot to compare sample purity generated after using a commercial cell fractionation kit (K = kit) versus the method reported here (NK = not kit). The kit was used to generate nuclear and cytoplasmic fractions only, these were compared for purity and yield against the method described by probing both fractions for (A) GAPDH and (B) Histone-H3. The C2C12 myotube fractions obtained from both methods were analysed on the same western blot (20 μg protein was loaded from each fraction).
In comparison with the described protocol, the yield obtained with the commercial kit as indicated by band density, was poor and only the cytoplasmic fraction appeared relatively pure. The kit is designed for more generic use with multiple cells and soft tissues and may explain the inability to generate a pure and abundant nuclear fraction from muscle. The main advantage of the kit was a slightly reduced processing time although an additional kit to purify a mitochondrial (and cytoplasmic) fraction would be required and this would increase the cost and time requirement.
In conclusion the method for subcellular fractionation described here is inexpensive, does not require an ultracentrifuge and was found to generate three relatively abundant subcellular fractions of reasonable purity. This method of subcellular fractionation could be combined with proteomics research wherein protein patterns of subcellular fractions could be mapped and characterized by 2D gel analysis and mass spectrometry.
Ivan Dimauro and Timothy Pearson joint first author.
Funding by the MRC (TP, MJJ) and University of Rome "Foro Italico" (ID, DC) are gratefully acknowledged.
ID and TP designed and carried out the cell culture and tissue experiments; MJJ and TP developed and supervised the experiments; MJJ, ID, TP and DC interpreted the data and wrote the manuscript. All authors read and approved the final manuscript.


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Das Schloss Parsow () ist ein Herrenhaus  im hinterpommerschen Parsowo in der heutigen Woiwodschaft Westpommern in Polen.

Geschichte
Ein Herrensitz in Parsow ist in einer Urkunde Kasimirs I. von Pommern von 1172 belegt. Die ursprüngliche Burg war vermutlich eine Fluchtburg, die durch Graben und Palisade geschützt war. Der Ort gehörte damals der Familie von Parsow, deren letzter Erbe der 1558 verstorbene Lorenz von Parsow war. Daraufhin wurde Jakob von Heydebreck durch Kurfürst Friedrich Wilhelm mit dem Ort belehnt. Unter der Familie von Heydebreck bestand ein zeitgemäßes Festes Haus.

Ab 1772 war die briefadelige Familie von Gerlach, vertreten durch Friedrich Wilhelm von Gerlach Besitzer des Guts. Dessen Nachfolger, Ludwig August Wilhelm von Gerlach, ließ 1782 den heute als Mitteltrakt erhaltenen Bau unweit des bestehenden Festen Hauses errichten. Bogislaw von Gerlach ließ nach 1860 dem Bau einen Nordflügel im Tudorstil zufügen. Die Nachfahren stifteten zur Erbregelung einen Familienfideikommiss. Der spätere Landrat Carl Heinrich von Gerlach.Heinrich Berghaus: Landbuch des Herzogtums Pommern. Teil III, Band 1.  W. Dietze Verlag, Anklam 1867, S. 400–401 (Online). übernahm die Gutsleitung. Ihm folgte zuerst der ältere Sohn, Leutnant Bogislav von Gerlach, dessen Karriere an der Ritterakademie am Dom zu Brandenburg begann. Dann folgte dessen jüngerer Bruder August von Gerlach, der eine Erwähnung im Jahrbuch der Millionäre erfuhr, ohne Angaben näherer Daten.

Um 1910 wurde ein Südflügel angebaut, um die drei Bauteile zu einem Gesamtbau zusammenfügen. Das Herrenhaus verfügte über eine Fideikommissbibliothek mit 4000 Bänden zur pommerschen Geschichte. Ende der 1930er Jahre, kurz vor Kriegsbeginn, beinhaltete das Rittergut Parsow eine Größe von 727 ha. Das dazugehörige Rittergut Schwemmin wiederum umfasste eine Fläche von 773 ha. Eigentümer war Carl August von Gerlach-Parsow (1883–1945), verheiratet mit Ruth von Bonin-Bottschow. Die drei Töchter heirateten in Adelsfamilien ein. Der einzige Sohn August-Tessen von Gerlach führte bereits ein Stadtgut Körlin und das Gut Trienke.

Bei der Besetzung durch die Rote Armee wurde das Schloss verwüstet, die Bibliothek verbrannte größtenteils. Heute ist das Herrenhaus restauriert.

Literatur
 Helmut Sieber: Schlösser und Herrensitze in Pommern. Nach alten Vorlagen. Weidlich, Frankfurt am Main 1959, S. 107 ff., 3. Auflage, Weidlich, Frankfurt am Main 1978, ISBN 978-3-8035-8012-2.
 Hans Friedrich von Ehrenkrook, Friedrich Wilhelm Euler, Genealogisches Handbuch der Adeligen Häuser 1954. B (Briefadel), Band I, Band 9 der Gesamtreihe GHdA,  C. A. Starke, Glücksburg/Ostsee 1954. S. 110–117.
 Gothaisches Genealogisches Taschenbuch der Briefadeligen Häuser. 1941. B. Zugleich Adelsmatrikel der Deutschen Adelsgenossenschaft. Jg. 33, Teil B. Adelige Häuser des seit Anfang des 15. Jahrhunderts bis zur Neuzeit nachgewiesenen deutschen Erbadels (späterer rittermäßiger Landadel, patrizischer Stadtadel, Reichsbriefadel, Landesbriefadel, Uradel und alter Adel nichtdeutschen Ursprungs, Offiziers- und Beamtenadel), Justus Perthes, Gotha 1940–10, S. 178–181.
 Handbuch für den Preußischen Adel. Band 2, Hrsg. Marcelli Janecki, E. S. Mittler & Sohn, Berlin 1893. S. 254–257. Digitalisat

Weblinks

 Schloss Parsow 1905

Einzelnachweise

Schloss in der Woiwodschaft Westpommern
Kulturdenkmal in der Woiwodschaft Westpommern
Gmina Biesiekierz
Bauwerk in Pommern
Erbaut in den 1780er Jahren
Historistisch überarbeitetes Bauwerk
Zerstört im Zweiten Weltkrieg
Rekonstruiertes Bauwerk in Polen
Parsow

redpajama_set_name: RedPajamaWikipedia


text:
Q: How to show Multinode tree picker value on its Parent Node Hi please need help how to render my multinode tree picker value (http://screencast.com/t/NTu1NqXsTmBz) to show it on its parent node named PORTFOLIO ..
i want the multinode picker value should be placed between the Portfolio and About section (http://screencast.com/t/F0Dnnv9a) but I don't know how yet i tried render it and still not work ..
here is the finish template : screencast.com/t/ar1zeZ43Pf6J
Node tree picker code:
@if (Model.Content.HasValue("mntp_pickContent"))
{
    var bannerListValue = Model.Content.GetPropertyValue<string>("mntp_pickContent");

     @foreach (var item in bannerListValue.Split(','))
        {
         var page = Umbraco.Content(item);

@page.contentTitle

@page.contentsubTitle

                                     More

        }

}

Parent Node Portfolio.cshtml
@{ if (Model.Content.HasValue("port_backImage"))
 {
     var bgport = Umbraco.TypedMedia(Model.Content.GetPropertyValue<string>("port_backImage"));

@Model.Content.GetPropertyValue("pageTitle")

@Model.Content.GetPropertyValue("subTitleText")

show all

                        @foreach (var portMenu in Model.Content.Children)
                        {

@portMenu.Name

                        }

 }
}

A: This all depends on the Templating Engine you are using.
If you are using MVC, then you need to add your MNTP logic to a partial view.
See the following link:
https://our.umbraco.org/documentation/Reference/Templating/Mvc/partial-views
If you are using WebForms, then you will need to add your logic to a Razor file OR alternatively a UserControl (.ascx) file. See the following links:
https://our.umbraco.org/documentation/reference/templating/macros/
https://our.umbraco.org/documentation/reference/templating/macros/Razor/
http://umbraco.com/help-and-support/video-tutorials/introduction-to-umbraco/developer-introduction/using-net-user-controls

redpajama_set_name: RedPajamaStackExchange


text:
The history of Captain Marvel
Captain Marvel was the brain child of Bill Parker and C.C. Beck. He was created in 1939 and first published in Whiz Comics #2 by Fawcett Comics in 1940. Captain Marvel's real identity is Billy Batson… a mere young boy. He was bestowed his powers by an ancient wizard named Shazam to become a champion of justice. Whenever Billy would speak the word "Shazam" he would be transformed in to the powerful Captain Marvel.
Captain Marvel was highly successful. DC took notice of Fawcett's success and decided that Captain Marvel was a rip-off of their own hero: Superman. They took Fawcett to court, but Fawcett settled, abandoning Captain Marvel to the clutches of DC.
DC published Captain Marvel under the title Shazam! thus thoroughly confusing anyone unfamiliar with Captain Marvel, as it appeared Shazam was the name of Billy Batson's alter ego.
Enter, me.
I never knew much about Captain Marvel, but he's a featured player in the current Infinite Crisis event. In Day of Vengeance (a title leading in to the Infinite Crisis) the Specter apparently went crazy, turned evil and started a campaign to destroy all magic (and thus, magic users)… including Captain Marvel himself.
Well upon reading this I decided I would check out an old issue (#9) of Shazam! that I had purchased from an antique store. In this issue, Captain Marvel battles Mister Mind and an army of worms. As if that wasn't stupid enough, the cover made me wonder if Archie Andrews had been transformed in to Captain Marvel. The comic was much worse than I was expecting. I could barely get through the issue. The comic was light-hearted and child-oriented and horribly boring.
These days, Captain Marvel maintains his innocent outlook, after all he's really just a child. Thus he isn't easily corruptible. He's like a less complex, more pure version of Superman. Although the Captain may be the same as he was in the past, the stories surrounding him have definitely darkened. In the end of Day of Vengeance, Shazam is killed by the Specter, and Captain Marvel (as Billy Batson) apparently gets the axe… apparently.
comic, Comics, DC, marvel, superman
3 responses to "The history of Captain Marvel"
Keith P. Stieneke says:
Infinite Crisis is one of the main comics I read write now since this mini-series/maxi-series started. I write a review of each one I read. I don't get all of the crossovers but I did just finish reading Day of Vengeance – Infinite Crisis Special and hope to post a review of it soon.
Tanisha says:
I think this is a helpful site. I am doing a superhero unit in class and I drew Captain Marvel. It gave me lots of information I needed. Thanks for all the help
Joe Louis says:
Tanisha:
Glad I could be of assistance… enjoy the site 🙂

redpajama_set_name: RedPajamaCommonCrawl


Resume Template ~ Free Truck Driver Resume Samples Beginner Sample Template Local Semi Examples For Students Marvelous Flatbed Truck Driver Resume. Cdl Truck Driver Resume Examples For Students. Free Truck Driver Resume Samples. Tow Truck Driver Resume Sample Download.
Marvelous flatbed truck driver resume dump examples pdf sample template cdl for teachers. Resume template truck driver samples velvet jobs flatbed sample pdf local semi examples professional for. Local semi truck driversume examples free professional sample template cover letter beginner tow.


Tolerates more sun and heat than most A. palmatum cultivars!
At last, a true weeping Japanese Maple! 'Ryusen' boasts a magnificent weeping habit with strong, pendulous branches bearing palmately divided leaves of bright green. It's a whole new look for the family!
Train 'Ryusen' to desired height until it forms a sturdy trunk, or let it weep over a wall, espalier it, or let it decorate your favorite arbor or arch. The cascading branches are wonderfully versatile, so you can find just the right spot in any landscape for this exciting introduction!
The foliage on this slow grower turns brilliant shades of golden-orange and bright red in autumn. More heat- and sun-tolerant than many A. palmatum cultivars, 'Ryusen' leafs out late in spring, thus avoiding late frosts. Expect it to reach 20 feet long and 6 to 8 feet wide over many years. Zones 5-8.
In northern zones, choose a location which receives full sun. Farther south, the location should be partially shaded. Bright sun helps produce the deepest colors in red cultivars. Soil should be rich and well-drained, high in organic content with ample moisture. Shelter from drying winter winds is desirable, particularly in the northern portion of its range.
Formative pruning is desirable, as is spur pruning for those cultivars with interesting bark coloration (A. 'Sango Kaku', for example). Such pruning should be done in late summer or early fall and, at the same time, any dead branches should be cut back to live wood. Water well during periods of drought, especially in the first 2 to 3 years after planting.


text:
6 Weeks of THE WORK | Day 17 | Rest
Who's looking forward to a rest day? I know I am.
I'm gonna be real with you today. It's really hard to avoid temptations on rest days. Something about not having to work out just makes me want some junk food! It's days like these that I'm really glad I've got Shakeology. It tastes like a treat, but it really helps me stave off those cravings. I thought that maybe it was just me, but it turns out that Beachbody conducted a survey of 2,769 people (including some independent Coaches like myself who drank Shakeology 5 or more times per week and exercised at least 3 times per week) and found that 81 percent of respondents felt like Shakeology helped reduce their cravings for junk food.*
Has Shakeology helped you cut back on snacking? What other benefits have you noticed from drinking Shakeology daily? Many customers who start drinking Shakeology typically feel all the results of drinking Shakeology within the first month. You can learn all about how Shakeology can positively impact your health in the Welcome Guide that was included in your first shipment. In the back of that guide there's even a Wellness Tracker where you can log how you feel on a day-to-day basis.
On that note, here's a new Shakeology recipe to add to your arsenal. I love the texture that oats and peanut butter add to a shake!
Strawberry Crumble Shakeology
Beachbody Guide to Nutrition pg. 45
I'm gonna be real with you today. It's really hard to avoid temptations on rest days. Something about not having to work out just makes me want some junk food! It's days like these that I'm really glad I've got Shakeology. It tastes like a treat, but it really helps me stave off those cravings. I thought that maybe it was just me, but it turns out that Beachbody conducted a survey of 2,769 people (including some independent Coaches like myself who drank Shakeology 5 or more times per week and exercised at least 3 times per week) and found that 81 percent of respondents felt like Shakeology helped reduce their cravings for junk food.
6 Weeks of THE WORK | Day 18 | Wk 3 | Endurance & Agility
6 Weeks of THE WORK | Day 16 | Wk 3 | Legs


text:
Pod elektritsjeskimi oblakami (Russisch: Под электрическими облаками; internationale titel: Under Electric Clouds) is een Russisch-Oekraïens-Poolse film uit 2015, geschreven en geregisseerd door Aleksej German. De film ging in première op 10 februari op het Internationaal filmfestival van Berlijn 2015 in de competitie voor de Gouden en Zilveren Beer.

Verhaal

Op een verlaten bouwsite staan onafgewerkte betonnen gebouwen, ijzeren constructies en autostrades. In de winterse mist en sneeuw dolen in deze surreële wereld een aantal figuren rond. Hun verhaallijnen verweven zich met elkaar.

Rolverdeling

|-
| Louis Franck || Pjotr
|-
| Merab Ninidze || Nikolaj
|-
| Viktoria Korotkova || Sasja
|-
| Tsjoelpan Chamatova || Valja
|-
| Piotr Gasowski || Djadja Bonja
|}

Prijzen en nominaties

Externe links

 Officiële website

Russische film
Oekraïense film
Poolse film
Dramafilm
Film uit 2015


first of all I cannot get in to stock recovery. I did the vol up home and power button, yes i let go of power when the logo appears and it stays on the samsung screen forever. part of a larger prob though. I was on GC 2.0. Yesterday I tried to get back to stock to do the update. nothing flashed right but I recalled whenever I try to flash AB it always brings me to stock recovery, so I did that. returned to stock. finally around midnight I got the update notification. I said yes and my phone hung on the samsung screen for the next half hour. even pulling the battery wouldnt get it to boot up so I had to reflash GC just to get my phone on so I could set my alarms for this morning.
Not sure where I went wrong. And I don't understand why I cant get in to stock recovery the way everyone suggests. I continued holding volup and home for 5 min each time. Doesnt seem to matter if I am stock or GC it just doesnt work. Thanks for your help everyone.


← Is the RV lifestyle for Us? Or maybe explore Central America?
Many people often start a side hustle while they are currently working to get some passive income, or side income that eventually can be their main income. Then Voila! Financial Independence. Sounds easy. It's not.
Research, hard work, follow through, possible failure, restarts, knock downs, get ups, work. It all comes down to hard work.
I have relatives who have been quite successful in commercial real estate rental. This is appealing because real estate is a physical tangible asset. If the space is rented (ignoring possible defaults) there is an ongoing monthly income stream. However there is maintenance required and therefore you need to stay near the real estate to really make this type of income work, and then it really isn't all that passive. This could be a viable option for us, as we would have help in finding a property and a possible tenant. It's just that the down payment would have to come from somewhere and I am hesitant to take it from my current investments as I am pretty loyal to my dividend holdings.
Writing online and earning blog income is something I find appealing, however with my limited website knowledge, this would require additional learning, training and would require time and effort. I am not afraid of either the time commitment or the effort. I just am not sure where to look for this training, and I don't want to sign up for a costly seminar where I don't learn what I actually need to know. And of the however many bloggers out there could I make mine stand out and gain followers?
Network Marketing is something that I am interested in learning about. In April, B & I are going to a training conference by Market America, where we will learn strategies to grow our business as well as more about the products they promote. I really see this being something we can use for future income. It's just a matter of getting out there. I know I will feel more confident and excited after receiving some training.
Finally there is Dividend Investing. I read almost daily about different investments and analysis. I am excited about this path of passive income and I am committed to regular savings and purchases. I think this is one of the best ways to earn passive income that is location independent. All you need is an internet connection, of course research and timing is crucial too. It is really exciting to receive those dividends in your account. It's a rush to put the orders through and to watch your account rise!
How do you feel about earning passive income? What's your strategy? How has it been working for you?
This entry was posted in Dividend Stocks, Financial Independence, Market America. Bookmark the permalink.
I think the options you mention all have strenghts and weaknesses, it comes down to what works best for you and what you're most comfortable with. I'm thinking that dividend investing and real estate are going to be the two areas I concentrate on.
There's many ways to earn passive and location independent income. So far, dividend investing has been great for me. We'll see if I sing the same tune if the market loses 50% of its value tomorrow.
I haven't found blogging to be a great source of passive income. I spend a great deal of my personal time on my blog and have realized very little monetary return on it. I think I've made less than $500 in the year I've been working on it. I have found it to be a great source of friendship building and mutual inspiration, but as a direct source of income I have found it to be absolutely horrendous and I cannot recommend it.


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Aramco's chief technology officer shares vision of circular carbon economy at ChemIndix
Published: 17 November 2019 - 7 a.m.
By: Martin Menachery
The 11th International Conference and Exhibition on Chemistry in Industry (ChemIndix) was held last week under the patronage of HRH Prince Khalifa bin Salman Al Khalifa, Prime Minister of the Kingdom of Bahrain.
The conference, organised by the Saudi Arabian International Chemical Sciences Chapter of the American Chemical Society, revolved this year around the theme of finding 'Breakthrough Solutions through Innovative Materials'.
Saudi Aramco has been supporting ChemIndix since its inception and this year was one of three diamond sponsors. The event focuses on developing breakthrough solutions to the challenges facing the energy and petrochemical industries.
Five main topics were discussed in three executive panels, five keynote speeches, 120 oral technical presentations, about 100 posters, and more than 35 think-tank sessions. The topics were on innovative materials in future cities and industries, digital transformation, breakthrough solutions in refining and chemicals, solutions in exploration and production, and on circular carbon economy and sustainability.
During a panel session on the concept of a circular carbon economy, Saudi Aramco's chief technology officer and the Executive Conference Advisory Board chairman, Ahmad O Al-Khowaiter, presented the view of the oil and gas industry.
Al-Khowaiter defined a circular economy as moving from a linear model, where materials are used and then disposed, to a more circular manner through the reduction of consumption, reuse, and recycle.
For circular carbon use, CO2 emissions from hydrocarbon combustion should be captured, reused, and recycled in other forms of energy to close the carbon cycle efficiently.
Al-Khowaiter emphasized the important role of oil and gas in powering the world economy through energy that sustains factories, mobility, and contributes to the growth and prosperity of societies. Leveraging renewables, he said, can ensure that these processes are carbon negative.
"Without that carbon-free energy, you can't recycle the CO2," said Al-Khowaiter. He noted that the circular concept presents an option to achieve a balance between societal needs, protecting the environment, and relying on the reduction and recycling of emissions only is insufficient.
The removal can be achieved via traditional methods such as forestry or industrial means. The goal, he said, is to achieve a balance between the carbon emitted and the amount reduced, recycled, and removed.
"This carbon balance is the natural order of things. So, taking our economies and making them aligned with nature just means restoring the carbon balance. That is what the Earth has been doing for billions of years, and we need to be able to emulate that," he said.
Al-Khowaiter highlighted Saudi Aramco initiatives to move toward the circular economy via reducing, recycling, and removing CO2. The company has undertaken a number of initiatives to reduce carbon emissions from its operations, ranging from the annual reduction of gas flaring since the 1980s, and committing to eliminate it completely by 2030, and to applying a comprehensive energy management program.
Beyond its operations, Saudi Aramco is collaborating with automotive engine manufacturers to maximize efficiency to reduce the emissions from using hydrocarbon fuels, he said. Additionally, the company is working with a number of partners to develop carbon capture and storage technologies for cars and trucks, and enabling the unloading of the CO2 at fuel stations for recycling into another energy form.
The latest development in this program is the successful testing of technology for heavy-duty trucks and achieving the capturing and storing capability of 45% of the CO2 emitted from the engine exhaust.
In terms of CO2 removal, Saudi Aramco is sequestering 800,000 tons of CO2 per year to enhance oil recovery in the 'Uthmaniyah field. The company is also examining different options for sequestering carbon, including directly removing CO2 from the air at an industrial scale.
Al-Khowaiter also spoke about Saudi Aramco's ongoing research projects that are designed to achieve advancements in chemistry and material sciences to enable the direct conversion of oil into chemicals through which CO2 emissions are reduced and efficiently captured and stored, achieving a level of recyclability of carbon.
As part of the conference, ChemIndix hosted a student hackathon where students from various universities from throughout the Kingdom competed to develop innovative and breakthrough solutions in three challenging areas — circular carbon economy (which was sponsored by Saudi Aramco), innovative materials, and the Fourth Industrial Revolution.
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Last week we provided an overview on the types of schemes that hackers use to gain access to business computer networks which they then hold ransom or mine for personally identifying information that can be used to commit fraud. The threat is real and it is costing businesses an incredible amount of money every year. So what can you do to protect your business from hackers? While there is no way to completely eliminate the threat of cyber attacks, these tips can help you stay safe. Learn more, below.
Cyber attacks can cause serious headaches for any business.
The first step in protecting your business from cyber attack is to have a plan in place. Your technology plan should support your overall business objectives while addressing the unique security challenges you face. Because technology is constantly changing, your plan should too: Best practice is to review your technology plan every 3 months in order to ensure that it is meeting your ever-evolving needs.
It is crucial that management teams outline proper procedures for how employees are expected to handle sensitive data. Keep in mind that many employees use their personal devices to access company information, so your policy should address what information can be accessed on personal phones, tablets, and laptops.
Many phishing schemes and malware infections can be prevented with a little common sense training. Do not assume that your employees know the importance of strong passwords and other internet safety practices. It is up to you to train them on how to protect themselves–and in extension, your business–from hackers.
Preventing cyber attacks takes some initial investment, including the costs for anti-virus software, hardware, professional development time, and more. These costs should be accounted for in your overall technology plan.
Sometimes these types of decisions are better made with an experienced advisor by your side. A tech advisor can help guide you on what type of training your staff would benefit from and the best technology investments you can make to protect your security. By working with an advisor, you can feel confident that your business is well-prepared to ward off the threat of attack.
This entry was posted on Thursday, September 1st, 2016 at 1:12 pm. Both comments and pings are currently closed.


Scott Morrison has today politely ignored former Prime Minister Tony Abbott's tips on how to keep the masses happy in the 2019 Federal Budget.
This comes as The Nightwatchman says the coalition, but mostly the Liberal Party, has been working towards the surplus it will deliver in Tuesday's budget since it was elected in 2013.
However, the soon to be unemployed Member For Warringah, Tony Abbott, says he worries Morrison might be pandering a bit too much to the working underclasses and atheists.
"Scott. I know times have changed and we have to look like we believe in climate change" he said.
The 2014 Australian federal budget was the federal budget to fund government services and operations for the 2014/15 financial year. The 2014 budget was the first delivered by the Abbott Government, since the Coalition's victory in the 2013 Australian federal election and put a big focus on making the poor pay for medical appointments, while also allowing concessions for ballet, and attempting to cut costs on mental health by replacing trained public school counsellors with Christian clergymen.
Abbott, who's is starting to fear his time in the top job has become so irrelevant that he might not even get a state funeral, says if there's one legacy he's willing to stand behind, it's that he made ballet cheaper for rich people in the North Shore.
However, while confident that his pentecostal Turnbull Lite will make the right decision regarding rich people hobbies and Christian counselling, he also had one big criticism.


#ICYMI: MASSCreative on TV! Matt Wilson talks with the BNN about raising awareness for arts funding.
Thanks to the BNN for helping us get the news out. We still have a lot of work ahead of us in the coming months. Let's keep working to ensure the next governor of Massachusetts is also a champion of the arts!


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5 Best Rules of Learning through Knowledge is about strengthening our power of knowledge. Many times we hear individuals, students and professionals talking about their learning and knowing abilities. Sometimes they experience extraordinary hurdles in their learning process. They discuss it with others and try to remove that obstacle. They learn from others insight.
It is human nature that it wants to experience everything by itself. It observes results by using its own solution. A person's exploring nature always force him to find out the solution by himself. This sometimes yield better results and sometimes not.
Everyday we read on Internet about other people's sharing of experiences of certain special situations. Their end results are acceptable as those are from personal experience. We can use them if we come across the same circumstances. To solve the problem from scratch sometimes become very time consuming. End results may or may not are positive.
Learning is a process of adapting information, which is theoretically or practically acknowledged. It is about accepting existing information, behaviors, skills, values, knowledge or preferences to put into action(s).
Knowledge is to get new information on facts, details, descriptions or skills acquire through experience(s) or education. It describes an awareness of something through theoretical or/and practical understanding.
Learning increases knowledge. Knowledge or a new information develops by sharing an individual's or a community's experiences with others. Knowing insights and practices of others shared by like-minded people increases knowledge.
Knowledge is a state of understanding which helps in forming the shape of action(s). Learning and knowledge reinforce each other in a cycle. In the process of learning knowledge, we polish our ability of comprehension.
Become an aggressive reader and listener.
Develop your habit of asking, 'ASK QUESTIONS'.
Share your information and experiences through discussions.
Help others by describing or explaining your information.
Remember, do not memorize some keywords from on going discussion.
These are a few steps that can help anybody in learning through knowledge. These are very simple rules, anybody can follow them. Improved knowledge will make our problem solving ability fast. This in return, will make less mistakes.
We must create a culture of knowledge and learning in our homes and in the society. Youngsters should learn to use their knowledge in solving their day-to-day problems. Less problems will give them chance to progress in their worldly and spiritual knowledge.
Your comments and insights are always WELCOME.


text:
New major Brazilian 'icon' planned
A proposal to completely transform the abandoned Cais Maua dock area of Porto Alegre in Brazil, which could include the construction of a selection of new homes, has been put forward.
Rio Grande do Sul's government has already appointed architects b720 Fermín Vazquez Arquitectos and Jaime Lerner Arquitetos Associades, as well as a consortium to design and deliver the planned £175 million Porto Alegre project.
The scheme is expected to include a new 1.6 mile stretch of waterfront, featuring a business and leisure zone for one of the 2014 FIFA World Cup venues. Additional plans include new build homes, a hotel, shops and office space in the derelict quays area.
A b720 spokesperson commented: "The transformation of Porto Alegre's waterfront is aimed at re-establishing a direct, human relationship with the Guaíba River, restoring the area's historical and cultural heritage, and creating a new icon for the city."
Brazil has become somewhat of a hotspot for buying property in recent years, with more overseas nationals, including Brits, now viewing the country as an attractive destination for a holiday home.


Policies, regulations, activities, and other information regarding the climate, physical landscape, and ecology of the United States.
Morris also wrote in his ruling that the Trump Administration "simply discarded prior factual findings related to climate change to support its course reversal." [post_title] => Keystone Pipeline Project Temporarily Halted After Federal Judge Rules Trump Administration Did Not Properly Study Impact [post_excerpt] => [post_status] => publish [comment_status] => closed [ping_status] => closed [post_password] => [post_name] => keystone-pipeline-project-temporarily-halted-federal-judge-rules-trump-administration-not-properly-study-impact [to_ping] => [pinged] => [post_modified] => 2018-11-17 23:16:36 [post_modified_gmt] => 2018-11-17 23:16:36 [post_content_filtered] => [post_parent] => 0 [guid] => http://factualuprising.com/?p=2693 [menu_order] => 0 [post_type] => post [post_mime_type] => [comment_count] => 0 [filter] => raw )  => WP_Post Object ( [ID] => 2680 [post_author] => 1 [post_date] => 2018-10-27 12:14:13 [post_date_gmt] => 2018-10-27 12:14:13 [post_content] => "Between 300 and 700 barrels of oil per day have been spewing from a site 12 miles off the Louisiana coast since 2004, when an oil-production platform owned by Taylor Energy sank in a mudslide triggered by Hurricane Ivan. Many of the wells have not been capped, and federal officials estimate that the spill could continue through this century." Taylor Energy has at times denied the leak and made efforts to keep it secret. "The spill was hidden for six years before environmental watchdog groups stumbled on oil slicks while monitoring the BP Deepwater Horizon disaster a few miles north of the Taylor site in 2010." Oil pipelines leak on average more than one time per day in the United States. The Trump Administration has proposed expanding offshore drilling areas to nearly all U.S. waters. [post_title] => 300-700 Barrels Of Oil Per Day Leaking Off Coast Of Louisiana Since 2004 From A Single Site Owned By Taylor Energy [post_excerpt] => [post_status] => publish [comment_status] => closed [ping_status] => closed [post_password] => [post_name] => 300-700-barrels-oil-per-day-leaking-off-coast-louisiana-since-2004-single-site-owned-taylor-energy [to_ping] => [pinged] => [post_modified] => 2018-10-27 12:14:13 [post_modified_gmt] => 2018-10-27 12:14:13 [post_content_filtered] => [post_parent] => 0 [guid] => http://factualuprising.com/?p=2680 [menu_order] => 0 [post_type] => post [post_mime_type] => [comment_count] => 0 [filter] => raw )  => WP_Post Object ( [ID] => 2678 [post_author] => 1 [post_date] => 2018-10-21 13:10:00 [post_date_gmt] => 2018-10-21 13:10:00 [post_content] => The Intergovernmental Panel On Climate Change (IPCC) of the United Nations issued a report on October 8, 2018 outlining the effects on global environments and ecosystems as global temperatures increase above pre-industrial levels. The IPCC was established in response to the Paris Climate Agreement, which seeks to "strengthen the global response to the threat of climate change by keeping a global temperature rise this century well below 2 degrees Celsius above pre-industrial levels and to pursue efforts to limit the temperature increase even further to 1.5 degrees Celsius." According to the report, "emissions of carbon dioxide (CO2) would need to fall by about 45 percent from 2010 levels by 2030, reaching 'net zero' around 2050. This means that any remaining emissions would need to be balanced by removing CO2 from the air."
Read the report here. [post_title] => United Nations Climate Change Panel Releases Report Stating Global Temperature Rise Of 1.5 Degrees Celsius Is Maximum Before "Irreversible Changes" To Global Ecosystems; Would Require 45% Reduction In Emissions By 2030 [post_excerpt] => [post_status] => publish [comment_status] => closed [ping_status] => closed [post_password] => [post_name] => united-nations-climate-change-panel-releases-report-stating-global-temperature-rise-1-5-degrees-celsius-maximum-irreversible-changes-global-ecosystems-require-45-reduction [to_ping] => [pinged] => [post_modified] => 2019-02-10 13:43:15 [post_modified_gmt] => 2019-02-10 13:43:15 [post_content_filtered] => [post_parent] => 0 [guid] => http://factualuprising.com/?p=2678 [menu_order] => 0 [post_type] => post [post_mime_type] => [comment_count] => 0 [filter] => raw )  => WP_Post Object ( [ID] => 2670 [post_author] => 1 [post_date] => 2018-10-06 14:11:20 [post_date_gmt] => 2018-10-06 14:11:20 [post_content] => The Environmental Protection Agency (EPA) issued a letter to the White House on Friday September 28, 2018 arguing that rules limiting mercury emissions from coal fueled power plants should be loosened because much of the benefit is not directly from reducing mercury emissions but rather other harmful pollutants that are also reduced because of this rule ("co-benefits").


text:
Fierce Female Fighting Forces
Afghanistan Commandos:
The Afghan army has trained women special forces troops to carry out night raids against insurgents, breaking new ground in an ultra-conservative society and filling a vacuum left by departing international forces Night raids have long been a divisive issue between Afghan president Hamid Karzai, who does not want foreign troops entering Afghan homes, and the US-led coalition that says the raids are essential to capturing Taliban commanders. Many Afghans, however, have complained that the house raids are culturally offensive. Having male troops search Afghan females is taboo. So is touching a family's Koran or entering a home without being invited. US Special Forces also had Female Cultural Support Teams previously in Afghanistan.
Read More at Express.co.uk
Turkish Special Forces Employs Females. Photo: Flickr
Turkish Special Forces:
Volunteers initially face a pass/fail written exam as well as physical tests. Those who pass the exams become special forces candidates. Adequate linguistic skills in at least one foreign language is a plus for admission. The candidates then have to complete a challenging training period which lasts around 2.5 years. Many drop out during this intense training period. Training encompasses fitness, obstacle, Martial Arts, close quarters combat (CQC), biological/chemical/nuclear warfare, parachuting, diving, sniping, demolition, counter-terrorism, search and rescue, stealth seek and destroy, reconnaissance and survival. Learning foreign languages is a part of the training.
Chinese Special Forces:
China established its first all-female special forces a few years ago. The unit has been shrouded in mystery until recently when media published photos of the female soldiers in training. The female soldiers follow the same training program as male counterparts, with no special allowances for being women. The unit has 45 members who are all female except for the male commander. These young women are aged from 18 to 26 and come from all backgrounds.
Read More at DramaFever
Korean Female Special Ops Photo: Flickr
South Korean Special Mission Battalion:
The 707th Special Mission Battalion is a special forces unit in the Republic of Korea Army Special Warfare Command. The unit is South Korea's primary counter-terrorist and quick reaction force. They are tasked with conducting urban counter terrorist missions, and constitute the Army's quick-reaction force for emergencies. The battalion's nickname is 'White Tigers'
The unit also has a small number of female special forces operatives. They are used in counter-terror operations where the presence of a woman is not seen as a threat to a terrorist.
Russian Female Paratroopers:
This series explores the lives of the women who serve in Russia's Airborne Forces, also known as the airborne infantry. These paratroopers are renowned for their desperate courage, stamina and fighting capacity. Do these ladies have what it takes to serve in this elite fighting force, where they must compete with the strongest and bravest of men.
Women In The Palestine Commandos
Palestine Female Commandos:
The female commandos are trailblazers in a still largely male-dominated society, set to become the first female members of the Presidential Guards, a Palestinian elite force of 2,600 men. Their inclusion is the result of gradual changes in the West Bank in recent years. For now, the female Guards are only being drawn from the West Bank. In the Gaza Strip, which is not under Abbas' control and is ruled by the Islamic militant Hamas, about 400 women serve in the 16,000-strong security forces. They have undergone some training, including in martial arts, but work mostly in administrative jobs, including as border control officers and an anti-drug unit.
Read More at ynetnews
Kurdish Female Soldiers:
To an ISIS militant, one of the worst things that could transpire in combat is not just being killed, but being killed by a woman. If this happens, ISIS members believe that they will go directly to hell. If hell exists, rest assured that they have been sent there by a number of Kurdish women.
Read more at All that is Interesting
IDF Females as part of Search and Rescue Photo: Flickr
IDF Search and Rescue:
The IDF National Search and Rescue Unit, under the Home Front Command, is a highly skilled force trained to execute special search and rescue missions, both in Israel and abroad. The unit was founded in 1983, and its' expertise is in rescuing people trapped under ruins. The unit is comprised primarily of reservists who are always on call, with prepared kits to enable immediate departure, and a small core of soldiers in mandatory service. In addition to the rescue teams, the unit employs doctors, engineers, mechanical engineering equipment operators and rescue dog handlers.
Read More at IDF Blog
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Both as advisor and then minister, Andrew Adonis was a key architect of Tony Blair's public service reforms in education, health, local government, policing and transport. He has deep and broad experience of the challenge of leadership, change management and policy implementation in complex organisations - and in difficult conditions.
Lord Adonis is author of books on parliamentary reform, the class system and the infamous poll tax – and was named 'Peer of the Year' in Channel Four's awards. After leaving the Cabinet he became Chairman of a cross-party think tank, working on ways to improve the effectiveness of government.