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This invention relates to an improved real-time scanning fluorescence electrophoresis apparatus for the electrophoretic analysis of fluorescently-labeled polynucleotide fragments. The apparatus is characterized by having an electrophoresis chamber containing an electrophoretic separation medium capable of accommodating multiple electrophoresis lanes arranged in a planar array, a fluorescence detector mounted on a translatable stage, a light source for exciting fluorescent molecules, and a computer for collecting data consisting of time, location, fluorescence wavelength and fluorescent intensity information. The improvements herein disclosed include, (i) using a spectral-array detector for detecting the emission light from the fluorescently-labeled polynucleotide fragments including the simultaneous detection of multiple fluorescent labels, and, (ii) a temperature control means to control the temperature of the electrophoretic separation medium during electrophoresis.
1. An improved real-time scanning fluorescence electrophoresis apparatus for the electrophoretic analysis of fluorescently-labeled polynucleotide fragments comprising:
an electrophoresis chamber including a front gel plate for containing an electrophoresis separation medium and defining a plurality of electrophoresis lanes;
a laser light source for creating a beam of laser light;
a bending mirror for directing the laser light into the electrophoresis chamber at right angles to the front gel plate;
a spectral-array fluorescence detector for detecting emission light from said fluorescently-labeled polynucleotide fragments including simultaneous detection of multiple fluorescent labels:
a translatable stage for mounting the spectral-array fluorescence detector and the bending mirror thereon for translating the bending mirror and the spectral-array fluorescence detector with respect to the electrophoresis chamber for scanning the beam of laser light and the spectral-array fluorescence detector across the electrophoresis chamber in a direction parallel to the front gel plate and normal to the direction of migration of the polynucleotide fragments; and
a temperature control means to control the temperature of the electrophoretic separation medium during electrophoresis.
2. The apparatus of claim 1 wherein an output of the spectral-array detector is processed so as to effect a virtual filter, such filter effecting the sampling of a plurality of discrete wavelength ranges.
3. The apparatus of claim 2 wherein the wavelength ranges of said virtual filter are 540, 560, 580, and 610 nm, each 10 nm wide.
4. The apparatus of claim 2 wherein the wavelength ranges of said virtual filter are 530, 545, 560, 580 nm, each 10 nm wide.
5. The apparatus of claim 1 wherein the spectral-array detector comprises:
(a) a diffraction grating to separate the emission light,
(b) a CCD array to detect a location and an intensity of the separated emission light,
(c) an optical arrangement to direct and condition the beam of laser light and the emission light in order to minimize scattered laser light reaching the detector.
6. The spectral-array detector of claim 5 having an optical arrangement comprising:
turning mirrors which direct the laser light to a desired location,
telescopic lenses which focus the laser light to a position within the electrophoresis chamber,
a bending mirror that directs the laser light at right angles to the electrophoresis chamber,
an aspheric collection lens that collimates the emission light,
a set of laser rejection filters that reduce extraneous laser light entering the detector, and,
a plano-convex lens which focuses the emission light at a desired location.
7. The apparatus of claim 1 wherein the temperature control means comprises thermally controlled front and back heat transfer plates which are in contact with a front and back face of the electrophoresis chamber.
8. The apparatus of claim 7 wherein the front and back heat transfer plates are made from coated aluminum wherein the coating acts to electrically insulate the heat transfer plates from an electrophoresis voltage.
9. The apparatus of claim 7 wherein the temperature control means comprises:
(a) a front heat transfer plate placed in contact with a front face of the electrophoresis chamber, wherein flow channels are formed within the front heat transfer plate including inlet and outlet ports,
(b) a back heat transfer plate placed in contact with a back face of the electrophoresis chamber, wherein flow channels are formed within the back heat transfer plate including inlet and outlet ports,
(c) a flowable heat transfer medium which is circulated through the flow channels in the front and back heat transfer plates,
(d) a pump to circulate the flowable heat transfer medium,
(e) a heat exchanger in which the flowable heat transfer medium can exchange heat with an ambient atmosphere,
(f) a computer for controlling a temperature of the heat transfer plates by controlling the flow of the circulating heat transfer medium,
(g) a temperature sensor in contact with the front and back heat transfer plates and electrically connected to the computer to relay temperature information to the computer.
10. The apparatus of claim 9 wherein said heat exchanger is replaced by a cooler wherein the cooler cools the flowable heat transfer medium below the temperature of the ambient atmosphere.
11. The apparatus of claim 9 wherein said heat exchanger is replaced by a heater wherein the heater heats the flowable heat transfer medium above the temperature of the ambient atmosphere.
12. The apparatus of claim 1 wherein the electrophoresis chamber comprises:
(a) a front plate and a back plate, where the back plate is defined as a plate through which the laser light enters the electrophoresis chamber,
(b) two spacers which serve to maintain a uniform separation between the glass plates, spaced so as to provide a chamber thickness of from about 0.1 to about 1.0 mm,
(c) a plate holder which can accommodate glass plates of varying lengths and which acts to support and secure said electrophoretic separation medium and wherein said plates are held firmly in place within the plate holder by clamps which keep the edges of the plates sealed to prevent said separation medium from leaking.
13. The apparatus of claim 12 further comprising a plate locating mechanism which optimally positions a detection region of the electrophoresis chamber with respect to a detection optics.
14. The apparatus of claim 12 further comprising a mirror coating applied to an inside-facing surface of the front plate so that the excitation laser light, after passing through the back plate and the electrophoresis chamber, strikes the mirror coating and is reflected back through the electrophoresis chamber, thereby exciting additional fluorophores whose light is then collected, resulting in an increased emitted light signal.
This invention relates to improved apparatus for performing electrophoresis, and more particularly to an improved real-time scanning fluorescence electrophoresis apparatus for polynucleotide fragment analysis.
Electrophoretic polynucleotide fragment analysis methods are used to characterize mixtures of polynucleotide fragments based on their migration velocity through a polymer network under the influence of an electric field, i.e. their electrophoretic mobility, in combination with single or multi-color fluorescence detection. Typically these methods are applied subsequent to amplification of the target polynucleotide using a method such as PCR, e.g. Mullis, U.S. Pat. No. 4,683,202. Examples of such methods include polynucleotide sequencing, e.g. Trainor, Anal. Chem., 62:418-426 (1990), restriction fragment length polymorphisim (RFLP) analysis, e.g. Watkins, Biotechniques, 6:310-319 (1988), and variable number of tandem repeat (VNTR) or microsatellite analysis, e.g. Ziegle et al., Genomics, 14:1026-1031. Each of these methods can provide valuable genetic information about the target polynucleotide.
Current electrophoretic polynucleotide fragment analysis systems are characterized by multiple electrophoresis lanes arranged in a planar array, e.g. a multi-lane slab gel, in combination with a real-time-scanning fluorescence detector, e.g. Hunkapiller et al., U.S. Pat. No. 4,811,218. Multiple lanes are used to increase the overall throughput of the analyzer. In order to collect data during the electrophoresis from multiple lanes, the optical detector system is scanned across the width of the electrophoresis chamber perpendicular to the direction of migration of the labeled polynucleotides. Preferably, multi-color fluorescence detection is used to increase the information density per lane, e.g. for DNA sequencing, four label colors are used, one color for each base. A light source, e.g. a laser, excites the fluorescent labels attached to the polynucleotide fragments, and multiple emission filters discriminate between labels having different spectral properties. In addition, a computer is used to collect data consisting of time, lane number, and fluorescence emission wavelength information, and transform it into useful information, e.g. DNA sequence.
A significant limitation on the speed and resolution of current polynucleotide fragment analysis systems is the ability to dissipate the Joule heat that is generated as a result of the electric current passing through the electrophoresis medium. Because of problems caused by Joule heating, current systems are limited to low, e.g. 25 V/cm, electrical fields, resulting in long analysis times, e.g. 8 hrs. Joule heating and the resulting temperature gradient across the gel can negatively impact the quality of the separation in two ways. First, because heat is generated throughout the electrophoresis medium but only dissipated at its' outside surfaces, a parabolic temperature profile is establish across the depth of the channel. Since electrophoretic velocity is a strong function of temperature, approximately 2% per �C., this temperature profile leads to a parabolic velocity profile for the migrating analytes. This spatial dependence of velocity causes a broadening of the migrating zones, leading to reduced separation performance. The extent of the temperature profile can be reduced by making the electrophoresis channel thinner, e.g. Brumley et al., Nucleic Acids Research, 19:4121-4126 (1991); Stegemann et al., Methods in Molecular and Cellular Biology, 2:182-184 (1991). Therefore, an automated system which incorporates thin electrophoresis channels would be desirable.
Second, if the average temperature of the electrophoresis medium becomes too high, the structural integrity of the medium can be compromised. In the case of polymer gel media, e.g. crosslinked polyacrylamide gels, the elevated temperature can lead to complete destruction of the gel. The average temperature of the electrophoresis medium can be controlled by increasing the rate of heat transfer from the electrophoresis channel to the surrounding environment. Therefore, a system which more efficiently transfers the Joule heat generated as a result of the electrophoresis to the surrounding environment would be desirable.
A further limitation on the speed and resolution of electrophoretic separations is the rate at which the detector can acquire data from fast moving analyte bands. The most desirable form of detection for polynucleotide fragment analysis would be simultaneous multi-color detection. However, current approaches, i.e. an indexable filter wheel in combination with a photomultiplier tube (PMT) detector, are not ideal because the filter wheel must be indexed rapidly enough to observe each color before it moves out of the detector region. This is problematic due to the high electrophoretic velocity of the polynucleotide fragments in high-speed systems. If a sufficient number of data points are not collected for each analyte band, e.g. 10 points per band, the ability to discriminate between adjacent bands is lost. One way to increase the rate of data acquisition for a multi-color system is to collect signals from all the colors simultaneously rather than serially. Therefore, a detection system which acquires all colors simultaneously would be desirable.
In light of the above, what was needed was an improved electrophoresis apparatus capable of accommodating high electric fields through enhanced heat dissipation characteristics and detector performance.
The present invention is directed to improvements to an apparatus for electrophoretic polynucleotide analysis, said improvements leading to increased throughput of the system. The improvements include (i) incorporating a spectral-array detector to increase the rate of data acquisition, and (ii) incorporating an improved means to control the temperature of the electrophoresis medium. The analyzer system of the present invention is comprised of, in combination.
An improved real-time scanning fluorescence electrophoresis apparatus for the electrophoretic analysis of fluorescently-labeled polynucleotide fragments of the type having an electrophoresis chamber containing an electrophoretic separation medium capable of accommodating multiple electrophoresis lanes arranged in a planar array, a fluorescence detector mounted on a translatable stage, a light source for exciting fluorescent molecules, and a computer for collecting data consisting of time, location, fluorescence wavelength and fluorescent intensity information wherein the improvement comprises:
(a) a spectral-array detector for detecting the emission light from said fluorescently-labeled polynucleotide fragments including the simultaneous detection of multiple fluorescent labels,
(b) a temperature control means to control the temperature of the electrophoretic separation medium during electrophoresis.
FIG. 1 shows an electrophoresis gel including sample loaded thereon.
FIG. 2 is a diagram of the illumination, collection and detection optics employed in the invention.
FIG. 3 shows a plate holder of the invention.
FIG. 4A, 4B, and 4C show a plate locating mechanism of the invention.
FIG. 5 is a diagram of a thermal control system used in the invention.
The term "polynucleotide" as used herein refers to linear polymers of natural or modified nucleoside monomers, including double and single stranded deoxyribonucleosides, ribonucleosides, α-anomeric forms thereof, and the like. Usually the nucleoside monomers are linked by phosphodiester bonds or analogs thereof to form polynucleotides ranging in size from a few monomeric units, e.g. 8-40, to several thousands of monomeric units. Whenever a polynucleotide is represented by a sequence of letters, such as "ATGCCTG," it will be understood that the nucleotides are in 5'-3' order from left to right and that "A" denotes deoxyadenosine, "C" denotes deoxycytidine, "G" denotes deoxyguanosine, and "T" denotes thymidine, unless otherwise noted. Analogs of phosphodiester linkages include phosphorothioate, phosphorodithioate, phosphoroselenoate, phosphorodiselenoate, phosphoroanilothioate, phosphoranilidate, phosphoramidate, and the like.
As used herein, "nucleoside" includes the natural nucleosides, including 2'-deoxy and 2'-hydroxyl forms, e.g. as described in Kornberg and Baker, DNA Replication, 2nd Ed. (Freeman, San Francisco, 1992). "Analogs" in reference to nucleosides includes synthetic nucleosides having modified base moieties and/or modified sugar moieties, e.g. described generally by Scheit, Nucleotide Arealogs (John Wiley, New York, 1980).
As used herein, the term "electrophoretic separation medium" refers to a material through which the polynucleotides are electrophoresed and which imparts a size-dependent electrophoretic velocity to the polynucleotides. Typically, such material is a porous network formed by linear or branched polymer molecules, or the like, e.g. crosslinked polyacrylamide.
As used herein, the term "electrophoresis chamber" refers to the container in which the electrophorertic separation is contained. Typically, this container is formed by two rectangular glass plates which are separated by thin polymer sheets, spacers, located between the plates at the edge regions of the plates. This is traditionally referred to as slab electrophoresis. When the electrophoretic separation medium is a rigid crosslinked gel, this format is referred to as slab gel electrophoresis.
FIG. 1 shows polynucleotide fragment samples (2) which have been labeled with one of several fluorophores loaded into loading wells (4) of vertically oriented slab gel (8), said gel mounted in the analyzer of the present invention.. The fragments are electrophoresed through gel (8) where they are separated based on their relative size. Following separation, the fragments pass through laser excitation and detection region (12) where the fluorescently labeled polynucleotide fragments are detected. The fluorophores emit light at a specific wavelength based upon the particular dye used, thereby facilitating the identification of each fragment.
After the polynucleotide fragments have been separated, they are detected by a simultaneous multi-color detection means. An important feature of the polynucleotide analyzer of the present invention is the "spectral-array fluorescence detector". As used herein, the term "spectral-array fluorescence detector" refers to a detector which employs (i) a means to spectrally separate the fluorescence emission light, such as a diffraction grating, or a prism, or the like, (ii) an array of detector elements sensitive to light radiation, such as a diode array, a charged coupled device (CCD) system, an array of photomultiplier tubes, or the like, (iii) an excitation light source, such as an incandescent bulb, an arc lamp, a laser, a laser diode, or the like, and (iv) associated optics capable of directing and conditioning both the excitation and emission light. The output of a spectral-array detector is light intensity as a function of array location, wherein the array location can be directly related to the wavelength of the light falling on that location. One example of such a detector is given by Karger et al., Nucleic Acids Research 19: 4955-4962 (1991).
One preferred method of treating the output of a spectral-array detector is to create a "virtual filter". As used herein, the term "virtual filter" refers to a method of manipulating data from a spectral-array detector such that a plurality of discrete wavelength ranges are sampled, wherein the location and bandwidth of each wavelength range can be dynamically changed using software. The virtual filter can mimic a physical interference or absorbence filter, however it has several important advantages. First, virtual-filters can be programmed to interrogate multiple emission wavelengths simultaneously, making possible the efficient multi-color detection of fast-moving analytes without the need to rapidly index a multiplicity of filters. Second, virtual filters can be programmed to detect a range of emission bandwidths. This is important because for any application, there exists an optimum bandwidth which results in an optimum combination of sensitivity and color discrimination: as the detection band width is made wider, the detector collects more light, thereby increasing sensitivity, however, at the same time, the broader bandwidth decreases the ability to discriminate between closely related colors. Third, virtual filters have essentially perfect transmission curves, i.e. the filter can discriminate between very closely related colors. Forth, the selected wavelength ranges of the virtual filter can be easily adjusted using software to match the characteristics of various excitation light sources and dye sets. Therefore, changing dye chemistries is a simple matter of changing the virtual filter with software, whereas a mechanical modification of the system is required when physical filters are used. Moreover, the selected wavelength ranges and band widths of the virtual filter can be changed dynamically, i.e. during the course of a run, to compensate for any spectral changes in the dye labels which occur during a run.
FIG. 2 is a schematic diagram of the light path in a preferred embodiment of the spectral-array detection system of the present invention. Preferably, the analyzer system of the invention uses a laser as a fluorescence excitation light source, e.g. an argon ion laser that emits a 40 mW, 0.67 mm diameter polarized light beam having intensity maxima at wavelengths of 488 and 514 nm. Light from laser (66) is reflected off of adjustably-mounted turning mirrors (68) which direct the laser light to the desired location. Telescope lenses (70) then reduce the beam diameter to approximately 100 μm, and bending mirror (72) directs the light into electrophoresis medium (104) at right angles.
Light emitted from the laser-excited fluorescent label is collected by aspheric collection lens (74) which collimates the light in the direction of the detector. The emitted light then passes around bending mirror (72) and through laser rejection filter (76), thereby reducing the level of scattered laser light entering the detector. Because the excitation laser light passes through the center of aspheric collection lens (74), a certain amount of laser light will be reflected directly back from the lens surface in the direction of the detector, causing unwanted background signal. Bending mirror (72), which is mounted in the center of laser rejection filter (76), acts to deflect this reflected light away from the collection path thus reducing the amount of reflected light entering the detector. The collected emission light then passes through plano-convex lens (78) which focuses the emission light at slit (80) mounted on the entrance to spectrograph (82). (Spectrograph (82) utilizes a 405 g/mm, 450 nm blaze grating with a dispersion of 17 nm/mm.) After passing through spectrograph (82), the light then falls onto CCD (90). The output signal from CCD (90) is transmitted to electronic computer (64) for subsequent data analysis and presentation.
To further increase the emission light signal and decrease background light scatter, a nonconductive mirror coating is applied to inside surface (102) of front gel plate (108). This surface reflects emission light back to the collection lenses rather than allowing it to be lost to the surroundings through the front gel plate. In addition, when the primary laser light strikes this mirrored surface it is reflected back through the gel, thereby exciting additional fluorophores resulting in more emission light. Furthermore, this mirrored surface decreases unwanted background light generated by the fluorescence of the front glass plate itself.
In order to interrogate all of the electrophoresis lanes on a real-time basis, the optical system described above, less turning mirrors (68) and computer (90), is scanned across the width of the electrophoresis chamber.
Another important feature of the present invention is the novel means used to mount the electrophoresis chamber onto the analyzer. Preferably, the electrophoresis chamber is formed by two glass plates separated by two spacers located at the left and right edges of the plates. The glass plates are mounted into a plate holder which acts to support and secure the glass plates along with an upper buffer reservoir in a convenient manner. See FIG. 3. The plate holder consists of rectangular frame (200) onto which is attached plurality of twist clamps (204). (Note that only one twist clamp is indicated in FIG. 3, as (84), in order to retain the clarity of the drawing.) When twist clamps (84) are in the horizontal orientation, they serve to secure the glass plates in the holder, and, when twist clamps (84) are in a vertical orientation, they allow the glass plates to be conveniently inserted or removed from the plate holder. The rectangular frame includes two locational registration notches (208) to insure the proper positioning of the plate holder in the analyzer. Beam-stop (212) is positioned so as to protect the user from direct exposure to the excitation laser light. The frame also includes two handles (202) to facilitate transportation of the plate holder assembly. The plate holder provides a means for detachably mounting upper buffer reservoir (216). A protrusion (228) on each side of upper buffer reservoir (216) is positioned such that when the uppermost twist clamps are in the horizontal position, the upper buffer reservoir (216) is forced against the front glass plate, thereby creating a liquid-tight seal between the upper buffer chamber and the front glass plate. Upper buffer reservoir (216) contains electrode (220) and electrical cable (224) for connecting electrode (220) to an electrophoresis power supply. The plate holder is designed to secure glass plates of varying lengths. For applications requiring less separation and/or a shorter analysis time, a shorter length would be used, and for applications requiring more separation and for which longer analysis times can be tolerated, a longer length would be used.
A further important aspect of the present invention is the plate locating mechanism. In order to efficiently collect the fluorescence emission light, the detection region of the electrophoresis chamber must be properly positioned with respect to the collection optics. Specifically, the detection region must be aligned such that the focal point of the collection optics is located within the separation medium, and not in the wall of the electrophoresis chamber. The plate locating mechanism insures that this positioning is reproducibly achieved. The mechanism will be described with reference to FIG. 4. When a thin electrophoresis chamber is being used, i.e. less than 0.2 mm, preadjusted locating pins (300) fit through notches (304) in back glass plate (308) and push front glass plate (312) against front tip (324) of locating pins (300). When a thick electrophoresis chamber is being used, i.e. greater than 0.2 mm, step-portion (320) of locating pins (300) is forced against back glass plate (312). Locating pins (300) are preadjusted such that the interior of the electrophoresis chamber is at the focal point of the collection optics. Glass plates (308 and 312) are forced against locating pins (300) by twist clamps (330).
While increasing the electric field across the electrophoresis chamber increases the speed of the electrophoretic separation, it also leads to increased Joule heat generated within the electrophoresis medium, which in turn can lead to destruction of the electrophoresis medium. To remove the heat generated by running "fast" electrophoresis, a temperature control mechanism (FIG. 5) has been developed. The temperature control mechanism includes a back heat transfer plate (400) against which back glass plate (404) is mounted to the instrument. Preferably, heat transfer plate (400) is made from coated aluminum. The coating acts as an electrical insulator to inhibit arcing between back glass plate (404) and the rest of the instrument. Within back cooling plate (400) are channels through which a flowable heat transfer medium can be circulated. Front heat transfer plate (408), also containing channels capable of being filled with a flowable heat transfer medium, is contacted with front glass plate (412). Pump (416) circulates the flowable heat transfer medium from reservoir (420) through front and back heat transfer plates (400 and 408). Heat is removed from the circulating flowable heat transfer medium by passing it through heat exchanger (424), thereby cooling the flowable heat transfer medium to ambient temperature. If superambient heating or subambient cooling of the gels is desired for a specific application, the flowable heat transfer medium passes through a heater or cooler (not shown) before flowing through the heat transfer plates. Active temperature control of the gel is effected by means of temperature sensors (430) mounted to the heat transfer plates in combination with computer (434) which regulates the temperature of the plates by controlling the flow rate of the flowable heat transfer medium through the heat transfer plates.
Although the invention has been illustrated by the foregoing description it is not to be construed as being limited to the materials employed therein but rather the invention is directed to the generic area as hereinbefore disclosed. Various modifications and embodiments thereof can be made without departing from the spirit or scope thereof.
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IFANS: Mapping K-pop's International Fandom maps the global landscape of K-pop fandom. It is composed of a series of qualitative studies that examine the attitudes and activities of global K-pop fans as well as a digital humanities project that aggregates information, curates digital artifacts and analyzes the global spread of K-pop through fan activity on the Internet.
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Girls' Generation (SNSD):: SONES
Girls' Generation (SNSD), a nine-member female K-pop group, is one of the most popular K-pop female groups as a result...

subset: commoncrawl


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Chesapeake Energy Corp.'s decision to cut directors' pay and other perks may save the company up to $1.65 million a year without addressing investors' concern that the board failed to rein in Chief Executive Officer Aubrey McClendon's borrowing and spending spree.
In 2009, investor Jeffrey Bronchick told directors of Chesapeake Energy Corp. that he was disgusted with their leadership.
Chesapeake Energy Corp. shareholders rejected two directors involved in a probe of Chief Executive Officer Aubrey McClendon's personal finances after slumping energy prices and overspending wiped out $2.6 billion in market value this year.
American International Group Inc., the bailed-out insurer, rebuffed a request from New York Comptroller John Liu to release workforce-diversity data that was made public by Goldman Sachs Group Inc. and MetLife Inc.
Chesapeake Energy Corp. shareholders issued a sweeping repudiation of the company's management yesterday by rejecting two directors and demanding more influence over the second-largest U.S. natural-gas producer.
Wal-Mart Stores Inc.'s June 1 annual meeting will feature Aerosmith and Cheap Trick as the warmup acts. The real entertainment starts when shareholders vote whether to re-elect the company's board.
Goldman Sachs Group Inc. and Morgan Stanley will rewrite their so-called clawback policies to make it clear they will reclaim pay from supervisors as well as from employees who take excessive risk or engage in improper conduct.
"Chile is our most favorite nation."
- Michael Garland on Oct 06, 2014
Pattern Energy's Garland Discusses Wind Power IPO


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Norway's oil fund, AXA invest in German properties
Published on Oct 10, 2012 5:32 PM
OSLO (REUTERS) - Norway's US$650 billion (S$800 billion) sovereign wealth fund and joint venture partner AXA France Insurance Companies have agreed to buy two properties in Germany for 784 million euros (S$1.24 billion) from the Royal Bank of Scotland, Norway's central bank said on Wednesday.
The properties are located in Berlin and Frankfurt and are used mainly for office and retail purposes. The two partners will each hold 50 per cent stakes in the properties, Norges Bank, which manages the wealth fund, said.
"It is in line with our strategy to build our real estate portfolio by initially investing in large, well-developed markets alongside partners with aligned interests," the bank said in a statement.


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18 Homemade Christmas Decorations: How to Make Christmas Decorations
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Top 12 Crafts for the X mas Party on a Budget
By: The Editors of FaveCrafts.com
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Throw a fabulous x-mas party on a budget with thrifty crafts for decorations, invitation and party favors. You can have fun this holiday season, entertain guests and keep that money in your wallet. Crafts are the secret to throwing a x-mas party on a budget. Whether it's school party, work party or a party at your own home for friends, you x-mas party will be the talk of the town.
X-mas Party on a Budget Table of Contents
X-mas Party Invitations
Noel Holly Card- Create a lovely Christmas card, perfect as a party invitation, in 4 easy steps.
Personalized Christmas Card- Jazz up plain store-bought cards using these techniques.
Merry Christmas Card- This lovely striped Christmas card is just right for a party invitation.
Christmas Gifts Card- You can turn a simple postcard and scrap paper into a Christmas card.
X-mas Party Decorations
Christmas Lights Garland- These pretty Christmas lights are made from construction paper. You can also use brown grocery bags and paint to create the lights.
Dazzling Christmas Centerpiece- Use leftover wrapping paper and garland to create these beautiful table accents.
Yuletide Table Accents- These elegant Christmas candleholders are made from clay pots.
Stackable Holiday Trays- Paint wood blocks and nested trays to create a tiered appetizer tray or a Christmas tablescape.
X-mas Party Favors
Elf Hat Gift Bag- Fill these bags with candy canes or other goodies for a simple and thrifty party favor.
Bottle Cap Christmas Tag- Give these cute Christmas tags out to your guests. They work as ornaments or gift tags.
Icicle Swirls- Use these easy beaded ornaments to decorate a tree during the party and then hand out as party favors at the end of the night.
Recycled Holiday Candle Wraps- Use old gift wrap (even crumbled gift wrap or the scraps from your Christmas wrapping) to create lovely wraps for candles.
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Member 21051
Jul 11, 2011
With all the cute ideas.....I can't beleive that is all you have to comment on. I'm sure nobody meant to offend you... :)
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Jul 10, 2011
I find the term 'X-mas' very offensive. It is Christmas.
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Lawry's Oven-Roasted Chicken
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- 1 roasting chicken
- Lawry's Seasoning Salt
- Pre-heat oven to 400 degrees.
- Clean chicken and sit in a roasting pan. Cover in salt. Put in hot oven and cook until done.
- When done, remove chicken from pan, and reserve drippings for gravy. While chicken rests let drippings separate from fat, and make gravy.
- Slice chicken and serve.
September 2012
This recipe is a personal recipe added by robinarroyave and has not been tested or endorsed by MyRecipes.
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Aberdeen LLC Storage Powers the Sony Digital Motion Picture Center
Sony Electronics unveils Aberdeen NAS storage at the Digital Motion Picture Center.
Culver City, CA (PRWEB) December 19, 2012
Server and Storage manufacturer Aberdeen LLC, in conjunction with Sony Electronics, Inc., is very pleased to announce that the Sony Digital Motion Picture Center (DMPC), located at Stage 7 on the Sony Pictures Studios Lot, is now fully networked with post-production workstations sharing Aberdeen NAS storage.
The DMPC is dedicated to the training and development of on-set and post-production workflows for the ever-growing world of 4K production. Since its opening in June, the DMPC has focused on Sony´s flagship camera, the F65, but will soon add the recently announced F55 and F5 CineAlta cameras to the Center. The addition of Aberdeen´s central storage system will network the DMPC´s post-production islands into a unified system that will emulate a real-world post house; enabling the exchange of both RAW and HD files from Sony´s growing line of large-imager cameras among the industries´ most popular applications.
"Aberdeen´s system gives the DMPC the ability to quickly share files produced by Sony´s cameras amongst the DMPC´s many workstations through a centralized storage system capable of moving any file size with the speed to meet the demands of multi-stream, real-time video manipulation," said Dan Perry, the Director of the DMPC. "Their technology is a perfect example of the types of workflow solutions that the DMPC was designed to highlight."
To handle Sony´s 8K and 4K RAW files with on-set and post-production workstations, Aberdeen fine-tuned its 5U, 96TB AberNAS N51L network attached storage (NAS). Aberdeen installed all of its hardware, including one of its own Stirling 474G Workstations, in a 21U rack and tied in all of the DMPC´s workstations with a complete 10 Gbit Ethernet and fiber infrastructure, which includes an Arista Networks10 Gbit Ethernet switch and a Hot Lava NIC card along with Emulex NIC cards in every workstation. The Aberdeen network delivers 10 Gbit and Gbit Ethernet connections to the application workstations through the switch with 40 Gbit of Ethernet bandwidth aggregated to the AberNAS storage device itself.
Sony 8K/4K Optimized, Aberdeen AberNAS N51L Specification Summary:
- Aberdeen 64 Bit Linux NAS Operating System
- Dual Intel Xeon E5-2667 2.9GHz Processors
- 192GB DDR3-1600 Total Main Memory
- 24 x 4TB Enterprise Level Hard Drives Configured RAID 10
- Mirrored 300GB 15K SAS 6G Operating System Hard Drives
- LSI 24 Port SAS 6G RAID Controller
These software applications (listed alphabetically) are being demonstrated at the DMPC by their respective developers on workstations in the Aberdeen network:
- Adobe Premiere
- Assimilate Scratch
- Avid Media Composer
- Blackmagic Design DaVinci Resolve
- Codex Vault
- Colorfront On-Set Dailies and Express Dailies
- Filmlight´s Baselight and Truelight
- Fujifilm CCBox
- MTI Cortex
- YoYotta
With the addition of Aberdeen´s system, the DMPC continues its mission to educate the motion picture and television industries on high-resolution image capture and workflow with weekly training sessions and to establish common practices for the fast-paced world of post-production. "As we migrate from film and tape acquisition to 4K and even 8K production, the DMPC continues to be a crucial facility for the evaluation and development of standards that will benefit the future of high-end production," according to the DMPC´s President, Alec Shapiro. Sony and Aberdeen look forward to working together to fill this vital role to lead the way in 4K production.
Aberdeen Media Contacts:
Larry Aguilar
Marketing Manager
Aberdeen LLC
562-903-1500 x153
Jack Tateel
Executive Vice President
Aberdeen LLC
562-903-1500 x157
Tom Di Nome
Senior PR Manager
Sony Electronics Inc.
About Aberdeen:
Aberdeen LLC is a leading manufacturer of servers and storage. Delivering exceptional performance, unparalleled reliability and outstanding value, Aberdeen´s award winning products are deployed every day by IT departments in many of the world´s largest organizations.
For the original version on PRWeb visit: http://www.prweb.com/releases/prweb2012/12/prweb10241130.htm


text:
Research Output for Huan Zhou
Note: The 2012, 2013, 2014, 2015 publications have not been audited.
Jump to: Publications | Grants and Consultancies | HDR completions
Huan Zhou may be from another institution or only recently joined Deakin University.
2012 External
Wu, Yuheng*, Zhang, Zili, Deng, Yong*, Zhou, Huan* and Qian, Tao* (2012) An enhanced multi-agent system with evolution mechanism to approximate Physarum transport networks, in Thielscher, Michael and Zhang, Dongmo (eds), AI 2012 : Advances in artificial intelligence : 25th Australasian Joint Conference Sydney, Australia, December 4-7, 2013. Proceedings, pp. 27-38, Springer Berlin Heidelberg, Berlin, Germany [B1]
DRO entry for this publication
2011 External
Zhou, Huan*, Zhang, Zili, Wu, Yuheng* and Qian, Tao* (2011) Bio-inspired dynamic composition and reconfiguration of service-oriented internetware systems, in Tan, Ying; Shi, Yuhui; Chai, Yi and Wang, Guoyin (eds), Advances in swarm intelligence : second international conference, ICSI 2011, Chongqing, China, June 12-15, 2011 : proceedings part I, pp. 364-373, Springer-Verlag, Berlin, Germany [B1]
Citation counts: Scopus - 1
DRO entry for this publication
Grants and Consultancies
No grants and/or consultancies found or audited – the 2014 HERDC financial return has not been completed or loaded as yet.
Higher Degree by Research Completions supervised by Huan Zhou
No completions found or audited.
- Legend:
- * Not a member of Deakin University at time of output.
DRO to publications collection last synchronised: Saturday 30th May 2015 10:04pm


text:
Current Filters
Ofna oversized flow-through black anodized cylinder head. ABC construction (Aluminum piston with a chromed brass cylinder). Rear exhaust. Ball bearing-supported crankshaft. Slide valve rotatable carburetor. Needle valve adjusted by a screwdriver. Pilot shaft. This kit includes one .21 Ofna Hyper engine w/slide valve carburetor. One brass collet (these hold flywheel in place). One header with seal, clamp and spring. One running and parts information sheets and one Hyper .21 decal sheet.
Fuel with 5%-30% Nitromethane (20% is recommended) and a lubricant content of 18% or more. Muffler. Glow plug (long plug without idle bar)


text:
Erin in the Soccer Championship in Coral Springs 2004...winners
A photojournalist a the South Florida Sun- Sentinel for over 36 years...Retired from the paper 5 years ago to do more of my own photography of Sailing, sports and photojournalist style weddings.
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My Bronica S2a has just arrived and after a test roll it seems that the focus is a bit out of line due to the aged and deteriorated foam that is supposed to press the ground-glass against the metal frame. I've done some research and replacing it seems pretty approachable even by a rookie like me, but I'm not that eager to start unscrewing things without a service manual.
The problem is that I can't find anything on the Internet except some for sale on Ebay etc.
I'm on a budget and I'd rather invest the few dollars I have left in films... Does anyone have a service manual for this camera and is willing to share?
Thank you!


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Lamb And Rice Casserole
|Olive oil
|1 Tablespoon
|1 1⁄2 Pound, cut into 1 inch cubes
|1 Medium, thinly sliced
|Chopped green pepper
|1⁄2 Cup (8 tbs)
|Uncooked long grain rice
|1 Cup (16 tbs)
|Dried oregano leaves
|1⁄2 Teaspoon
|Dried basil leaves
|1⁄4 Teaspoon
|1⁄4 Teaspoon
|1⁄4 Teaspoon
|1⁄8 Teaspoon
|Beef bouillon cubes
|2 Cup (32 tbs)
|Canned tomato sauce
|8 Ounce (1 Can)
Heat the 1 tablespoon olive oil in a skillet over moderately high heat on a conventional surface unit.
Add meat cubes and cook until lightly browned on all sides.
While meat is browning, in a deep, 2 1/2 quart, heat-resistant, non-metallic casserole dish, heat onion slices and green pepper in the 1 tablespoon olive oil in microwave oven on full power for 4 to 5 minutes or until vegetables are tender.
Stir once during cooking period.
Add browned lamb cubes, rice, oregano, basl, paprika, salt and pepper to onion mixture.
Mix thoroughly.
In a 4-cup, heat-resistant, non-metallic measuring cup, combine the bouillon cubes and water and heat on full power for 4 to 5 minutes or until boiling.
Stir to dissolve bouillon cubes.
Add tomato sauce to bouillon.
Pour over meat mixture in casserole, cover, and heat on simmer for 35 minutes or until meat is tender.
Any excess liquid will be absorbed if casserole is allowed to stand 5 to 10 minutes


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Brain function at issue in Erskine case
A Stanford University research scientist explained to a jury in the death penalty trial of a South Bay murderer last week why learning how the brain functions is important to the case.
"You're going to have to decide whether this is an issue involving biology or an issue of good and evil," said Robert Sapolsky, who also is a professor of biology and neurology.
Sapolsky is one of several neurologists, psychologists and other experts called by lawyers representing convicted child killer Scott Erskine to testify whether their client's brain was seriously damaged when he was 5 years old.
They have asked the jury to spare Erskine's life because, they say, his criminal acts were at least partly the result of something he could not control: mental problems caused by that brain damage.
The jury convicted Erskine last month of molesting and torturing 13-year-old Charlie Keever and 9-year-old Jonathan Sellers, who were abducted off a bike trail near the banks of the Otay River on March 27, 1993, and strangled.
Their partially clothed bodies were found two days later. DNA evidence analyzed again in 2001 linked Erskine to the crimes.
All death penalty juries are instructed to consider many things before deciding if the punishment should be death or life in prison without the chance of parole.
Erskine's attorneys, Deputy Public Defenders Larry Ainbinder and Juliana Humphrey, have been focusing their efforts in two areas:
-Whether Erskine was under the influence of extreme mental or emotional disturbance when he killed.
-Whether Erskine's capacity to appreciate the criminality of his conduct was impaired as a result of mental disease or defect.
If jurors find either of those factors to be true, they could be weighed in favor of recommending that Erskine be imprisoned for life rather than sent to death row.
Two experts testified last week they believe that when Erskine was hit by a car as a child, it severely damaged the front part of his brain. That injury, combined with being raised in a dysfunctional family, created a situation in which he was unable to "put the brakes on aggression," one of the experts testified.
Erskine has a lengthy criminal history of sexual assaults for which he was frequently incarcerated. He committed his first sex crime when he was 10 before continuing to rape and assault boys, girls and women when he was not locked up.
Several defense experts testified that because of the brain injury, Erskine's ability to inhibit his "sexually sadistic" behavior was compromised.
Deputy District Attorney Valerie Summers aggressively challenged many of the defense witnesses. They acknowledged that predicting how people will behave based on psychological tests and brain scan results – the basis for most of their testimony – is difficult and inexact.
On Friday, Summers called to the witness stand Lori Behrens, a woman who was Erskine's roommate in San Diego when the boys were murdered.
While defense experts described Erskine as being unable to control his desires because of his brain damage, Behrens testified that Erskine was a likable, normal man who was always respectful and never violent.


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INDEPENDENCE TOWER Apartments728 N Jennings Rd, Independence, MO 64056 View On Map (816) 257-1400
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Researching the following surnames: Stier, Coffey/Coffee, Waldron, Jackson, Patman, Cumberbatch, Christiani and McDavid, Garms, Goff, Walling, Lillienthal, Merrell/Merrill, Stout, Elliott, Smith, Cook, Johnson, Bruce, Moore, Greer, Watson, Boney and Wells.
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Barbara Campbell
RE: Bidant Moore Coffey #54571050
Greetings! I am a direct descendant of her Father, Jesse Moore Sr. So she would be my Aunt several generations back. I can also claim kinship to all of the of the Coffey family from Mulberry, Caldwell county, NC. My Grandfather was Bruce Coffey. Should you wish to pass Biddie to me my # is 47122870. Thank you, Barbara Campbell
Added by
Barbara Campbell
on Apr 20, 2015 6:15 AM
Becki Christian Depew
Thomas J Coffey family #41647374
By any chance are you related or know anyone who is related to the Thomas J Coffey (Coffee) family? I am interested in his wife Rhoda C Christian Coffey #4164731. There is speculation that she is related to my Christian family out of Morgan County, Missouri. Do you know if anyone has a copy of her death certificate or other documents that may list her parents or place of birth, or perhaps any oral tradition as to her Christian family?
Becki Christian Depew
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Becki Christian Depew
on Apr 15, 2015 3:59 PM
Craig Meyers
RE: Meyers/Piske Transfer
Thanks Margaret. Don't worry, all transfers are in good hands. Whatever works for you is fine for me so we will just let things be. Nice to hear from relatives of the first marriage!
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Craig Meyers
on Sep 11, 2014 6:16 PM
Craig Meyers
Grave Transfer
Could you transfer another grave ownership to me? Her name is Albertina Meyers Piske. Memorial number 45681102 . I am her great grandson and I would like to finish the links to the rest of the family.
Thanks much,
Added by
Craig Meyers
on Sep 06, 2014 9:30 PM
Craig Meyers
Grave Transfer
Could you transfer a grave ownership to me? His name is Ludwig Theodore Friedrich "Lewis" Meyers. Memorial number 45680761. I am his great grandson and I would like to finish the links to the rest of the family.
Thanks much,
Added by
Craig Meyers
on Sep 06, 2014 5:39 PM
Kenny Lockett
Peter Hinrich Garms
Greetings-I am new here as I just discovered this site. Peter Garms was my great great grandfather. His daughter Sophia Elizabeth Garms Lockett was my great grandmother. Her and Wyatt Coffey (Doc) Lockett sixth child, William Vernon Lockett, was my grandfather. Looking forwARD TO CORRESPONDING WITH YOU.
Kenny Lockett
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Kenny Lockett
on Jul 01, 2014 1:05 PM
RE: Forrest Goff #119832946
Very Welcome.
J Miller
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on Jun 03, 2014 10:08 AM
Dennis Harmsen
Edit Requests
Just a note to let you know there are some Edit Requests that I sent to you, that are still pending. Do you have any Edit Requests in your Edit Tab that are still waiting to be processed/accepted? Thanks.
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Dennis Harmsen
on Apr 29, 2014 10:20 AM
Lu Ann
Stier connection
I am the great-great-great granddaughter of Amelia Ann Hilton and William Henry Stier (their daughter Ann married my great-great grandfather, Champion Harrison, both of them are also buried in Loudon Park). I met another one of Sarah Burgess Stier's descendants at her grave last year. Do you have any information on Amelia Ann Hilton? It would appear she was born in New Market, but I've searched everywhere (including the Historical Society, Hall of Records), and I cannot trace her beyond her marriage to William Henry in Frederick. I am fixated on finding out more about her. Thanks.
Lu Ann Marshall
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Lu Ann
on Feb 10, 2014 7:38 AM
Beverly Witmer
RE: Harry Lilienthal obituary
You are welcome. Found it while searching for an obit for someone else.
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Beverly Witmer
on Feb 07, 2014 6:16 PM
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From Here to There with Some Godbehere's
Three Generations of Godbehere Men
by Written & Compiled By: Tiffany S. Hogg Pankau
This book is a Family History of Godbehere men.
I am the granddaughter of Shirley Jean (Godbehere) Hogg. I love genealogy and learning of my ancestors!
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Gear Review
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My BSF!
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Pakistani nuclear deals and international security.
A year ago, international monitors had unearthed the central role Pakistan played in assisting Tehran in Iran's covert uranium enrichment programme for almost two decades. Only recently, Colonel Gaddafi's son, in an interview with a British newspaper, made public Pakistan's role in Libya's clandestine pursuit of a nuclear route to developing weapons of mass destruction (WMDs). Strangely now, the official denial mode has been replaced by putting the blame on the individual scientists engaged in selling the WMD technology, alleging that they did it on their own for personal gain by keeping the government of Pakistan in the dark.
The Islamists in Pakistan consider Khan as a national hero and do not want any action against him. His confession is a smart deal between him and the army. Khan's confession speaks for itself. He absolves the army of any role in the proliferation, and he was pardoned by Musharraf for his misdeeds.
Lieutenant General Khalid Kidwai, who heads Pakistan's Strategic Planning and Development Cell, described Khan as the mastermind who was involved in an elaborate and wholly unauthorised smuggling network that started in 1989 and was brokered by a host of middle-men. According to Kidwai, Khan told investigators that the assistance to the three countries was not meant to make money, but was a gesture of support for other Muslim countries. Kidwai claimed that Khan had, in a twelve-page signed document, admitted that he had transferred nuclear technology to North Korea, Iran and Libya.
It is hard to believe that helping Shiite Iran was ideologically compatible with the Pakistani establishment's Sunni ideology. The reason for transferring centrifuge designs to Iran reportedly between 1987 and 1991 was probably money. The North Korean deals were downright commercial. By the late 1980s, Pakistan had a nuclear capability, but no missiles. It bought the Nodong missile from North Korea and renamed it Ghauri. The probable rationale in the Libyan case was personal corruption. Thus, these three different considerations seem to have inspired the sale of Pakistan's nuclear secrets.
Pakistan's laws on terrorism and extremist groups remain opaque. The government claims to be tackling terrorism and recent arrests seem to have provided crucial information about al-Qaeda's plans to attack four targets in the United States, and Heathrow airport in London. Yet it has taken no substantial steps towards restricting the institutionalised extremism that permeates parts of society. Indeed, many Pakistanis argue that President Musharraf is following the pattern of the country's previous military rulers in co-opting religious extremists to support his government's agenda and to neutralise his secular political opposition. Whatever measures have so far been taken against extremism have been largely cosmetic to ease international pressure.
Everyone knows that Pakistan's nuclear programme is controlled by the army. Since its inception, Pakistan's nuclear programme has been squarely under army supervision. A multi-tiered security system was headed by a lieutenant general with all nuclear installations and personnel kept under surveillance. Thus, nothing could have happened without the army's knowledge. Pakistani officials profess to be shocked to find out that Pakistani scientists have been hawking their expertise for fifteen years to a list of clients that includes Libya, Iran and North Korea.
It is difficult to believe that Khan did this without knowledge either of successive Pakistani governments, or of the military. As Pakistan was a recipient of US technology, one can only wonder how the US was unaware of what was going on. The American investigative journalist Seymour Hersh reported that Washington was aware that since 1997 Pakistan had been helping North Korea build the bomb by sharing sophisticated technology, warhead design information and weapons testing data.
Khan's confessions should not come as a surprise to the international community. Soon after Pakistan's military defeat by the Indian armed forces in Bangladesh's liberation war in 1971, then prime minister Zulfiqar Ali Bhutto declared that Pakistan should acquire nuclear weapons and related delivery systems to match India's military capacities.
While a number of senior Pakistani nuclear scientists opposed Bhutto's nuclear ambitions, Khan showed a willingness to do what was needed. He initiated clandestine processes of acquiring material and technology for the manufacture of nuclear weapons in Pakistan. Between 1972 and 1974, Pakistan had persuaded Libya, Saudi Arabia and Iran to fund its nuclear weapons programme. By 1977, Pakistan went into high gear to become a nuclear weapons state after having collected the basic materials necessary to produce nuclear weapons. The first Pakistani nuclear device was tested at China's testing site at Lop Nor in Sinkiang province in 1987. By 1992, Khan openly confirmed that Pakistan was a nuclear weapons-capable state.
Khan had the protection of elements of the Pakistani military, which oversees the nuclear programme. His nuclear salesmanship left a wide trail of evidence that should have prompted action long before this.
* Iran: The UN inspectors' probe into Iran's nuclear programme revealed it had received centrifuges for enriching weapons-grade uranium from the nuclear black market. Khan is suspected of having supplied these in addition to weapon-related designs. Investigators charged Khan with transferring such nuclear components between 1989 and 1991. Pakistani officials say he met Iranian scientists in Karachi and in Malaysia.
Khan's travels to Iran in the late 1980s were common knowledge. The army chief of staff at the time, Aslam Beg, openly advocated a strategic alliance with Iran against the US at the time of the Gulf War. US officials raised concerns about a nuclear deal, but were assured it was not true.
* Iraq: UN inspectors found documents in Iraqi files in the early 1990s of nuclear aid offers from Khan, supposedly turned down by Baghdad.
* North Korea: Between 1997 and 2002, Khan made 13 trips to North Korea, secretly swapping centrifuge design and components for uranium enrichment in return for many of Pyongyang's Nodong missiles, later renamed the Ghauri. Much of the cooperation began in the 1990s when Pakistan purchased ten Scuds from North Korea.
* Libya: After Libyan leader Colonel Gaddafi agreed to dismantle Libya's WMD programme, the US found warhead designs and other components sold to them by Khan and his team through the grey market for $50 million. Senior Pakistani leaders now confirm that Khan transferred nuclear technology to Libya between 1991 and 1997.
Pakistan was acquiring nuclear weapons capacity not only to counter India's conventional military superiority. Bhutto had told the Organisation of Islamic Conference (OIC) countries in Lahore in 1974 that Pakistan's bomb would be an 'Islamic bomb' and could be the foundation for Islamic countries acquiring strategic military capacity to counter other nuclear weapons powers. This was an indication that Pakistan would be willing to supply nuclear material and know-how to other Islamic countries.
The second point to note is that Pakistan's nuclear capacities were built with the support of a number of Western European countries and the US, which were signatories to the Nuclear Non-Proliferation Treaty (NPT). The question arises why the US and Western powers did not monitor and counter Pakistan's nuclear weapons aspirations in the late 1970s and 1980s.
The fact of the matter is that Western powers were fully aware of Pakistan's nuclear weaponisation programme but turned a blind eye because of their desire to utilise Islamabad to resist the Soviet incursion into Afghanistan. In the eighties, Pakistan fully cooperated with the West in fighting the Soviet forces in Afghanistan. This connivance by the West continued more or less until September 11, 2001. It was only after the direct terrorist attack on the US that Pakistani nuclear weapons activities came under scrutiny and were subjected to investigation.
The US Administration appears to be concerned about the security of Pakistan's nuclear arsenal as well as possible leakage of sensitive nuclear know-how to the terrorist groups, which is simply reinforced in Congressman Joseph Crowley's recent warning that a threat from Islamic militants from Pakistan should not be downplayed as these terrorists might get access to that country's nuclear weapons. Lately, international pressure on Islamabad has compelled the Pakistani authorities to interrogate the nuclear scientists involved in this racket.
US President George W Bush, speaking publicly for the first time about Pakistan's nuclear black market network, has cited it as a failure of the international safeguard regime, but he avoided comment on the pardon granted by Musharraf to Khan. This operation represents a serious danger of nuclear proliferation. He said: 'The government of Pakistan is interrogating the network's members, learning critical details that will help them prevent it from ever operating again', and 'Pakistan's President has promised to share all the information he learns about the Khan network and has assured the United States that his country will never again be a source of proliferation'. He made a strong case for new international efforts to combat the spread of WMDs, saying the most dangerous threat before the world is the potential for terrorists or rogue nations to use chemical, biological, radiological or nuclear weapons in a surprise attack. Washington has made it clear that it considers Islamabad a major nuclear proliferator. Washington also views Pakistan as one of the unstable states in the region. It recognizes that power in the wrong hands in Islamabad would constitute a serious threat to its interests. Washington fears that after Musharraf it could be the deluge in Pakistan.
The fact remains that the dangerous actions of Khan are an eye-opener for the world. The rogue nature of his actions in league with those who either ignored or supported his actions needs an inquiry. This issue affects regional security and has international implications in terms of nuclear security and safeguards. A matter of continuing concern should be as to what stage the countries to which Pakistan exported nuclear material and nuclear know-how had succeeded in the development of nuclear weapons. Failure to uproot this nuclear network poses a serious threat to the global community.
Nations have at least three motives for building nuclear weapons programmes.
* The desire to be a world power, based on the belief that a nation unable to defend itself against the full range of possible dangers cannot be a world ower. Such a nation will both acquire nuclear weapons and strive for the capability to reach any potential adversary. Anxious to preserve their special status, these states are least likely to engage in proliferation except, as in Russia, due to a collapse of discipline. They are also least vulnerable to sanctions because the other world powers value their cooperation on other subjects. India is in this category.
* States that feel threatened by neighbours with large populations or greater resources may see in nuclear weapons a means to pose unacceptable risks or to create a deterrent against threats to their survival. This is especially the case if the powerful neighbours have nuclear weapons. Such states could be kept from developing nuclear weapons only by a credible guarantee from existing nuclear powers, which is unlikely to be extended and even less likely to be believed. Pakistan is in this category.
* Nations determined to wreck the balance of power in their regions and that see in nuclear weapons a means with which to intimidate their neighbours and discourage outside intervention. North Korea and other so-called 'rogue states' are in this category.
History demonstrates that the United States has always perceived the proliferation of nuclear weapons as a danger not only to world stability but also to its own national interests. It stood to reason that if one nation possessed the means to unleash the devastation wreaked on Hiroshima, then several such nations would represent a concerted threat, particularly if those nations were not friendly to the United States.
Attempts to control the spread of nuclear weapons, however, were met with problems. While America was undoubtedly committed to the notion of non-proliferation, the extent to which it was able to practise non-proliferation was limited by the fact that non-proliferation was an issue rarely isolated from Washington's other concerns. When it came down to having to decide between preserving today or guarding tomorrow, the logical and only decision Washington could make was to preserve today. After all, if today's interests were not protected, tomorrow would have little chance of being realised. Consequently, the long-term objective of non-proliferation was often compromised in order to ensure the short-term nature of American strategic interests. The result was the emergence of a host of nations whose strategic significance was deemed important enough to warrant exemption from measures designed to prevent the attainment of nuclear weapons' capability.
The 1998 nuclear detonations by India and Pakistan underscored the fact that, by putting strategic concerns before non-proliferation objectives, the US had practised a short-term and limited policy of securing America's current interests at the cost of tomorrow's security.
Pakistan's nuclear pursuit cannot be separated from India's. Pakistan's nuclear programme has been largely reactive to India's. India too has bought, borrowed and procured by dubious means nuclear materials and technologies from sources as diverse as the UK, US, USSR/Russia, Norway, France and Canada. The source of the plutonium used in India's 1974 test was spent fuel from CIRUS, a 'research' reactor of Canadian design, to which the US donated heavy water. India reneged on its promise to use CIRUS solely for 'peaceful' purposes. To escape legal censure, it called the 1974 test a 'peaceful nuclear explosion'. This was a form of cheating too.
Then there are the states of the former Soviet Union, which have hundreds of unemployed nuclear technologists and unscrupulous businessmen willing to trade in forbidden material. Therefore, the danger of nuclear proliferation will always be there unless there is universal nuclear disarmament. Israel's nuclear capability is a known secret. Nor can an exclusive club of the US, Russia, China, France and Britain be allowed to remain nuclear powers, while others are denied the same right. There are fears in some circles in Pakistan that the powerful non-proliferation lobby in the US is not going to let Musharraf off the hook so easily. They believe the US will exploit this situation to gain joint custody and regular inspection of Pakistan's nuclear programme. The leading defence analyst, Lt General Talat Masood, does not believe the US will crack down on Musharraf but, he argues, in an interview with India Today (February 16, 2004), the real concern now is that 'the worst-case scenario will be constantly portrayed--that there is a growth of militant groups outside government control which may seize Pakistan's nuclear assets'.
It is expected that Pakistan will be under pressure to repair its reputation and will have to take some confidence-building measures relating to its nuclear setup. Pakistan would in no circumstances permit foreign inspectors to enter the country and monitor its nuclear weapons or civil nuclear facilities, General Musharraf has said. 'This is a very sensitive issue. Would any other nuclear power allow its sensitive installations to be inspected? Why should Pakistan be expected to allow anybody to inspect?', he asked.
He further said:
Pakistan had no intention of freezing its nuclear weapons programme. It was self-sufficient and would not require the import of more material or designs from abroad. We will never stop our nuclear and missile programme. That is our vital national interest. It is totally indigenous now. Whatever had to be imported and procured has been obtained.
In a similar tone, the Foreign Minister, Khurshid Mahmood Kasuri, while addressing at a recent Pakistani foreign policy seminar in Islamabad, said:
Pakistan will never compromise on its nuclear capability. It will not roll back or freeze its nuclear programme. The nation should rest assured that the nuclear capability, which enjoys national consensus, is in safe and professional hands.
The Khan episode has left the Pakistani nation insecure and demoralised. Its credibility has been seriously damaged in the international arena. The government has decided to pass the necessary legislation to prevent any transfer of nuclear technology. Pakistan's nuclear programme is now headed by two lieutenant generals (previously one), with all nuclear installations and personnel kept under the tightest possible surveillance. In addition, it has now renewed its energies on running down al-Qaeda. Hopefully, the non-proliferation regime will see merit in these actions.
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|Property type
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|3 - 6 nights
Fantastic apartments in the perfect spot!
We have two luxury apartment located by the sea outside Lisbon to Cascais and has stunning seaview from all rooms in both apartments.
The 190 m2 apartment has four bedrooms, very large living room/kitchen in open solution and with a veranda towards the sea . Two toilets and the apartment is fully equipped.
New for 2015, the 120 m2 apartment has two bedrooms, large living room/kitchen in open solution and with two verandas towards the sea . Two toilets and the apartment is fully equipped.
The sandy beach is just outside the house, and if you do not want to swim in the sea, there is a pool and a garden belonging to the apartment. The apartment has a car place in the garage and apartment complex is guarded / front desk 24/7.
If you do not want use the car the train station is at 5 min walking distance and it takes 20 minutes to go to the centre of Lisbon.
Around the apartment there are a number of cafés and restaurants and 5 minutes walking distance is cosy Parede centre with shops and market for fish, fruit and vegetables, etc. every day.
The apartment complex also has an external fully equipped gym, with 5-10 classes per day, for example, yoga and spinning. The gym also has an indoor pool, jacuzzi and sauna. Avalible for special rates for guests.
If you are 5 or less you have up to 20% discount (except high season June-Aug) for the 190 m2 apartment.
Please note that the photos is from both apartment but in the text below its marked to which apartment the photo belongs to.
We are a Swedish Brazilian married couple. We spend most of our time in Portugal and in Sweden.
We cannot think of a better place, in Europe. The apartments are fabulous and you are close to everything. It's a quite noble area and not very touristic. On the cafes, restaurants and small stores around they quickly gets to recognize you and takes good care of you. You have the gym, pool and beach on you doorstep and if you want to go to the center of Lisbon or Cascais, you take the train, less than 20 min to Lisbon and 10 minutes to Cascais center. Close to the house you have plenty of restaurants and cafes and the prices are much less than on the touristic areas. Then of course, for us that like to keep moving, you have the beach of carcavelos to walk or run but best of all is the gym club and spa in the same building, its so great.
Nice amenities. Nice view. Nice area just outside Lisbon.Great location
We very much enjoyed our stay in this apartment. It was extremely large and spacious and safe enough for our one year old child to play in. It was perfectly clean and very well maintained. The kitchen was excellent and we cooked meals in it most nights. The apartment has stunning views of both the beach and the local town. The area itself is perfectly pleasant with excellent beaches within walking distance. We didn't hire a car but all amenities were within walking distance including supermarkets and pharmacies. There are also some excellent Portuguese restaurants near by. We visited Cascais several times by the train. The station is a short walk from the apartment with a regular and very cheap train service. Overall our stay was excellent and we would recommend this property to our friends.
Fond memories of an excellent holiday.
The apartment was really perfect for our needs. Very well planned with good space for all of us, well equiped and in very good condition. The flat was clean and very well prepared upon arrival.
The owner had prepared good documentation that was very helpful, not only for the flat itself, but also with good recomendations on activities, restaurants and surroundings. I would recomend this apartment strongly: Perfect location with beaches and central Lisbon in convinient distances, good conditions and very high standards (kitchen, equipment, Internet, TV etc). After hectic days on the beach surfing or playing in the waves, we all looked forward to a couple of hours with relaxing time either at the pool side or just "hanging around" in a very nice and welcoming apartment.
The apartment is placed in the perfect location with a stunning sea view. It is big enough for 2 families without feeling crowded. The kitchen was well equipped so no problems cooking all the lovley fresh food you buy. The balcony has plenty of chairs and is facing the sea.
Supermarket is only 5 minutes walk away. Plenty of cafes close by if you want to eat lunch out. Beach is around the corner and very clean and nice with great facilities. Also plenty of cafes at the beach for ice cream breaks!
The apartment is very light and fresh. It almoust seems brand new!
Wifi worked perfectly which is very convenient.
We are very happy with our stay and would definitely come back!
We stayed in this apartment last June for one week with my children and a friends family. It is a fantastic apartment in a really nice location. We enjoyed the closeness to Carcavelos Beach for surfing and also the really nice pool area The village Parede is genuine and really nice - not as touristic as Estoril and Cascais. Still you have a big grocery store and restaurants and cafes within nearby. The apartment is large and spacious. Everything is new and fresh! We will come back again!
I stayed in this apartment last August for two weeks with my husband, my children and my brother and his family. It is a wonderful apartment and a very well maintained building. We enjoyed the pool area (the kids loved it) and the beach is just a few steps away. They also have great cafes just outside the building and a big grocery store is within walking distance. The apartment is very large, airy and great for entertaining. Everything feels new and fresh! If you are considering a vacation rental in this area, this is the place! We can't wait to go back again!
The property is located along the seaside and the beaches are 1 min distance with restaurants and beach activates such as surf and snorkeling. There are quite many nice cafés closely and the center of Parede is very nice with its daily market with fresh fish, fruit and vegetables. With a car you are 15 min away from Lisbon center and 10 from Cascais center. Of you prefer the train, there is a station just 5 min walk from the house and the train takes you to Cascais in 10 min and to Lisbon in 20 minutes.
|Rental Period
|Minimum Price
|Additional Nights
|Weekend Night
|Monthly *
|Event Price
Middle Season S 2015
29 Mar 2015 - 5 Jun 2015
3 night minimum stay
for 3 nights
|Notes: Rates are based on 5 guests; add €15 per night, per additional guest
High season 2015
6 Jun 2015 - 28 Aug 2015
6 night minimum stay
for 6 nights
Middle Season H
1 Sep 2015 - 31 Dec 2015
4 night minimum stay
for 4 nights
|Notes: Rates are based on 4 guests; add €15 per night, per additional guest
My Standard Rate
3 night minimum stay
for 3 nights
|Final cleaning
|Linen and towels
|Refundable Damage Deposit
Payment policy: 20% with booking. Another 20% until 60 days prior arrival. Last 60% until 30 days prior arrival.
Cancellation policy: 20% until 60 days prior arrival. 40% until 30 days prior arrival. If new guests book: 20%
* Approximate monthly price. Actual pricing will depend on the days of the month you stay.
Please Note: The owner of this property accepts payment in the original currency specified (EUR), before use of conversion tool. The conversion tool is provided for your convenience only. The owner may accept other currencies if specifically agreed in advance.
Learn more about this holiday rental at the owner's website:


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Gal Gadot is an Israeli actress and model. She is most known for her role as Gisele Yashar in Fast & Furious and Fast Five. Ms Gadot won the Miss Israel title in 2004 and went on to represent Israel at the 2004 Miss Universe beauty pageant.
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Barcelona, Spain
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So glad I chose to do this. Our guide was a mannerly young man who knew the area well. Such a gorgeous spot and so wonderful to be in this pristine and natural setting. Very special to see the 2 little villages as well. Wonderful, crisp,sweet mountain air was just what I needed after navigating through a foreign city. I rate this tour 5 stars.
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Results 1 to 3 of 3
Thread: Pinup
November 8th, 2005 #1
Hide this ad by registering as a memberNovember 8th, 2005 #2
That seems like an unnaturally harsh start to her rib cage below her left arm. The left arm itself has the feel of being pasted on: It's not anchored correctly at the shoulder. Her breast is really large, even for a pinup girl.
Oh...and give her some fingernails!
The beachball is really well done and I think the reflection works.
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Her left hand is really awkward and the pinky finger is a little nub. And her bikini bottom looks painted on where it rides up her ass, maybe some wrinkles in the cloth would remedie it this does have promise though.
Strange coincidence that every man whose skull has been opened had a brain.


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Does the G series still scratch the lens???
Started Sep 19, 2013 | Questions thread
Dan Hudson OP
Regular Member • Posts: 289 • Gear list
Thanks for your answers. I have been through the lens scratch problem with a brand new G10/11 camera. Don't need that anymore or the extra effort to protect the so called lens protectors. Will just stay away from the G series. I submitted the question about self lens scratching because if I was not satisfied with my purchase of the Nikon P7800, I might again seek a G series camera. You answered my question so I will wait on delivery of the P7800 to see if it is all I hoped for. I guess the next step will be a Sony RX100. Not my first choice, though. Thanks again....Dan
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Alison Davis is founder and CEO of Davis & Company. Alison sets the strategic direction for Davis & Company and leads the development of new services and solutions. Alison is editor of the iBook, 49 ways to improve employee communications (2013) and co-author of the books, The Definitive Guide to HR Communication (FT Press, 2011), and Your Attention, Please (Adams Business, 2006). Alison is a former online columnist for The New York Times and frequently writes articles for business publications, including The Conference Board Review, Communication World and PR Strategist. Alison leads sessions for national organizations and associations. She is a member of the International Association of Business Communicators, the Public Relations Society of America and the Communication Leadership Exchange. Alison earned her B.A. from Douglass College, Rutgers, The State University of New Jersey.


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Copper Canyon and Batopilas
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El Gran Payaso
Joined: Apr 2006
Location: San Antonio
Oddometer: 8,883
Originally Posted by
Thanks, Tricepilot. I enjoyed your posts. I ride to Big Bend, the River Road, and Presidio every year and have often thought about Copper Canyon. What percentage of the roads are dirt and is a dual sport required? Did you guys camp or stay in motels/hotels? Any more pictures and info. would be appreciated. Thanks.
There are lots of folks to ride to Copper on "road" bikes. The area is a gold mine of twisties and fabulous vistas. However to make the decent to Batopilas I would not want to go on anything but a straight-forward dual. I spent way more than half the time standing on the pegs - Fastway footpegs by the way - and was GLAD to have the capabilities of the bike underneath. I didn't see a single roadie going in or coming out of the canyon itself.
Motodiscovery did not originally plan to go to Batopilas on this particular off-road training trip. I advised the owner, Skip, that it was my opinion that most if not all folks who were making the effort to get to the Canyon AND to avail themselves of this type of training absolutely wanted to get to Batopilas. In my mind it would be like going to Colorado for climbing training and never getting vertical. So, Skip added a day free of charge to the trip and down to Batopilas we all went.
Like I said, you can have a blast on your road bike all over the Sierra Madres, the Canyon area, and more. But if you want to plunge down the canyon and along the cliff (see my first pic of this thread) - and you truly want to enjoy the ride, be safe, and not beat up your bike - go dual.
For places to stay, in Creel I recommend the Pueblo Viejo, but there are all sorts of places to stay in Creel. In Batopilas, there are several places that are talked about on other threads. I haven't heard of Batopilas filling up. Beware that there are trains with American RV tourists (and their RVs) pulling into Creel, and some of them have vans or pickups take them to overnight in Batoplias. Right now there aren't enough bike riders alone to fill the hotels, but I can't say what impact the RV gang has there. If you're the type that needs confirmation of accomodations, it wouldn't be hard to arrange.
You might want to push past Presidio, on to Chihuahua, and then down to Creel on your ride, then make arrangements to get to Bato somehow, just to see it. Leave the bike topside. Or, rent a dual sport and do the whole trip yourself to the bottom. It's truly one of the "rides of a lifetime".
As you've read in my thread, I did the River Road, Davis Mountains etc. leaving Mexico on this trip. I see why you go there all the time - west Texas alone is absolutely beautiful.
So go tell your wife you're adding another bike to the garage!
Collective Guide to Riding in Mexico
Off-Road Durango to Mazatlán
The Treasure of the Sierra Madre
Into the Blue at Quintana Roo
On Mexican Time
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Experts Question Impact of Newspaper Endorsements
Sen. John Kerry, D-Mass., leads President Bush in terms of endorsements, but in an age when many people get their news from the Internet and cable news channels, experts question the influence of newspaper endorsements on voters.
As of Monday, 128 newspapers' editorial pages had endorsed Senator Kerry, while 105 newspapers supported President Bush for a second term, according to Editor & Publisher magazine. The papers backing Senator Kerry account for 16.9 million readers compared to 10.9 million readers of papers for the president.
More significantly, Senator Kerry picked up a total of 35 papers that had backed Mr. Bush in 2000, including Michigan's Flint Journal, Washington's Seattle Times, New Mexico's Albuquerque Tribune and the Morning Call of Allentown, Pa. Meanwhile, President Bush picked up four papers that had supported then Vice President Al Gore four years ago, including Colorado's Denver Post, Michigan's Macomb Daily, Pennsylvania's York Daily Record and the Herald-Journal of Spartanburg, S.C., E&P reported Oct. 25.
Some media experts, however, said newspaper endorsements are less likely to sway large numbers of voters, who get information from a variety of sources.
"Editorial endorsements are dinosaurs. The vast majority of the public don't read editorials," Larry Sabato, political science professor and head of the University of Virginia's Center for Politics told UPI, a wire news service.
"The handful of people who read editorials already know for whom they are going to vote. They are either reading it for reinforcement or they are reading it because it's part of a newspaper.
While they were useful in the 18th century, today "there are thousands of options for interested news consumers, voters, and this really is horse and buggy. People make up their own minds. People who are going to vote in this election know what the stakes are," Sabato said.
As media critic Howard Kurtz of the Washington Post noted in an online discussion Oct. 25: "Even editorial page editors will tell you that they don't believe their endorsements move many votes."
In fact, several large national papers — including USA Today, the Wall Street Journal and Los Angeles Times — do not traditionally endorse presidential candidates and will likely abstain from issuing formal endorsements this year.
Of the nation's larger newspapers that did issue endorsements, Senator Kerry captured two: the New York Times and the Washington Post, with circulations of more than 1 million and almost 773,000 respectively. The larger papers to endorse President Bush this year include the Arizona Republic in Phoenix, San Diego Union-Tribune, Houston Chronicle, Dallas Morning News and Chicago Tribune, a paper with a circulation of nearly 579,000.
"There is much the current president could have done differently over the last four years. … But for his resoluteness on the defining challenge of our age — a resoluteness John Kerry has not been able to demonstrate — the Chicago Tribune urges the reelection of George W. Bush as president of the United States," the paper's editorial board wrote Oct. 17.
More interesting, said Kurtz, were endorsements from newspapers located in battleground states, such as Florida and Pennsylvania. In states where the race is particularly close, campaigns hope these endorsements could have some bearing on voter decisions.
When Florida's Orlando Sentinel on Oct. 24 published its endorsement of Senator Kerry, the campaign quickly issued a press release trumpeting the news, highlighting that the paper had not backed a Democratic presidential candidate in nearly 40 years.
The Sentinel wrote in its editorial: "This president has utterly failed to fulfill our expectations. We turn now to his Democratic challenger, Sen. John Kerry, with the belief that he is more likely to meet the hopes we once held for Mr. Bush."
Meanwhile, President Bush received endorsements from several larger newspapers in the battleground state of Ohio, including the Columbus Dispatch and the Cincinnati Enquirer, which praised the president's leadership in the war on terrorism.
Kathleen Hall Jamieson, dean of the Annenberg School for Communications at the University of Pennsylvania, noted that endorsements are less important to readers than they are for the campaigns, which often use them as proof of their superiority over their rival.
"The effect of the editorials doesn't come out of people reading them, they come out of the ads by the candidates saying 'I've been endorsed,'" Jamieson recently told UPI.


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Acquisition of Daum by Financiere Saint-Germain Creates Luxury Group of Complementary Products With a Distinctive Savoir-Faire
Financiere Saint-Germain now embarks on a program of decisive growth for some of the world's most respected and appreciated names in decorative luxury products and objets d'art for discriminating interior designers, consumers and collectors worldwide.
Financiere Saint-Germain is controlled by the family of Mr
Founded over 130 years ago in
According to
In terms of jobs, Financiere Saint-German has committed to maintain the industrial site at Vannes le Chatel, near Nancy. Mr. Prosper adds, "I am convinced that luxury requires French manufacture, with savoir-faire that marries tradition and innovation. Every factory has its own expertise and craftsmen of great quality. We must now build a distribution network that offers all of the group's brands in one place."
Daum, the luxury crystal maker renowned for producing one-of-a-kind originals, recently unveiled its "Vintage Collection" in
"Daum will continue to enthrall the upscale collector's market with its revolutionary color-changing crystal and new artist showcases. At this month's America's Polo Cup in
"It's an honor to represent Daum especially now as it joins such a distinguished family of luxury products through this brilliant acquisition by Financiere Saint-Germain," said
SOURCE Daum