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Michael De Leon
Michael A. De Leon is a graphic designer by day, but wears many hats in his off time. In 2004, after several years of covering the Spurs, he started Project Spurs, a Spurs team fansite as an outlet to provide content to Spurs fans, while continuing to write for himself and learning the ins and outs of online publishing and web design. He has since built a writing team and started a popular weekly Spurs podcast called the Spurscast.
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by: Jeff Garcia According to Euro-step.net, the San Antonio Spurs will sign Australian center Aron Baynes to a multiyear deal. Baynes, who was playing for Slovenia team Union Olimpija, is a 260 lbs., 7-0 center and played at Washington State during college (2005-2009). Click HERE to read the entire article and watch the video [Read More]


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Synthetic media formulations are disclosed for use with blood preparations intended for in vivo use, including synthetic media formulations to be employed in conjunction with the photodecontamination of platelets.
1. A synthetic platelet storage media substantially free of glucose, comprising an aqueous solution of:
45-120 mM sodium chloride;
5-15 mM sodium citrate;
20-40 mM sodium acetate;
20-30 mM sodium phosphate;
and a psoralen at a concentration greater than 0 and below its maximum solubility in water.
2. The synthetic platelet storage media of claim 1 wherein said media has a pH of approximately 7.2.
3. A synthetic platelet storage media substantially free of glucose, comprising an aqueous solution of:
approximately 86 mM sodium chloride;
approximately 10 mM sodium citrate;
approximately 30 mM sodium acetate;
approximately 26 mM sodium phosphate;
and a psoralen at a concentration greater than 0 and below its maximum solubility in water.
4. The synthetic platelet storage media of claim 3 wherein said,media has a pH of approximately pH 7.2 and an osmolarity of approximately 300 mOsm/Kg.
5. A synthetic platelet storage media, substantially free of glucose, comprising an aqueous solution of:
approximately 86 mM sodium chloride;
approximately 10 mM sodium citrate;
approximately 30 mM sodium acetate; and
approximately 26 mM sodium phosphate.
The present application is a continuation of patent application Ser. No. 08/692,444, Aug. 5, 1996, now U.S. Pat. No. 5,908,742 which is a continuation of patent application Ser. No, 08,338,039, Nov. 14, 1994, abandoned, which is a continuation-in-part of patent application Ser. No. 08/072,485, filed Jun. 2, 1993, now U.S. Pat. No. 5,459,030 which is a continuation-in-part of patent application Ser. No. 07/926,477 filed Aug. 7, 1992, now abandoned which is a continuation-in-part of U.S. application No. 07/844,790, filed Mar. 2, 1992 U.S. Patent No. 5,288,605 issued Feb. 22, 1994.
The invention generally relates to synthetic media for use with blood preparations intended for in vivo use, including synthetic media used in conjunction with the photodecontamination of platelets.
Whole blood collected from volunteer donors for transfusion recipients is typically separated into its components: red blood cells, platelets, and plasma. Each of these fractions are individually stored and used to treat a multiplicity of specific conditions and disease states. For example, the red blood cell component is used to treat anemia, the concentrated platelet component is used to control bleeding, and the plasma component is used frequently as a source of Clotting Factor VIII for the treatment of hemophilia.
Ideally, all blood cell preparations should be from freshly drawn blood and then immediately transfused to the recipient. However, the logistics of operating a blood donor center preclude this possibility in the vast majority of cases. Transfusions are needed day and night and it is difficult, if not impossible, to arrange for donor recruiting at unusual hours. Consequently, modern blood donor centers must use stored blood products.
In the United States, blood storage procedures are subject to regulation by the government. The maximum storage periods for the blood components collected in these systems are specifically prescribed. For example, whole blood components collected in an "open" (i.e., non-sterile) system must, under governmental rules, be transfused within twenty-four hours and in most cases within six to eight hours. By contrast, when whole blood components are collected in a "closed" (i.e., sterile) system the red blood cells can be stored up to forty-two days (depending upon the type of anticoagulant and storage medium used) and plasma may be frozen and stored for even longer periods.
While red cells are stored in the cold, Murphy and Gardner, New Eng. J. Med. 280:1094 (1969), demonstrated that platelets stored as platelet-rich plasma (PRP) at 22� C. possessed a better in vivo half-life than those stored at 4� C. Thus, more acceptable platelet concentrates could be transfused after storage at room temperature. Until recently, the rules allowed for platelet concentrate storage at room temperature for up to seven days (depending upon the type of storage container). However, it was recognized that the incidence of bacterial growth and subsequent transfusion reactions in the recipient increased to unacceptable levels with a seven day old platelet concentrate. Platelet concentrates may now be stored for no more than five days.
One might believe that it is a relatively simple matter to keep the blood preparation sterile during the manipulations needed to concentrate the platelets. After all, blood bags used for platelet concentrate preparation are in themselves sterile, as are the connected satellite bags. However, bacteria can be introduced by at least two different means.
First, if the donor is experiencing a mild bacteremia, the blood will be contaminated, regardless of the collection or storage method. Adequate donor histories and physicals will decrease but not eliminate this problem. See B. J. Grossman et al., Transfusion 31:500 (1991).
A second, more pervasive source of contamination is the venepuncture. Even when "sterile" methods of skin preparation are employed, it is extremely difficult to sterilize the crypts around the sweat glands and hair follicles. During venepuncture, this contaminated skin is often cut out in a small "core" by a sharp needle. This core can serve to "seed" the blood bag with bacteria that may grow and become a risk to the recipient.
Indeed, many patients requiring platelet transfusions lack host-defense mechanisms for normal clearing and destruction of bacteria because of either chemotherapy or basic hematologic disease. The growth of even seemingly innocuous organisms in stored platelets can, upon transfusion, result in recipient reaction and death. See e.g., B. A. Myhre, JAMA 244:1333 (1980) and J. M. Heal et al., Transfusion 27:2 (1987).
The reports assessing the extent of contamination in platelets have differed in their methods, sample size, and bacterial detection schemes. D. H. Buchholz et al., Transfusion 13:268 (1973) reported an overall level of platelet contamination of 2.4% when a large (>1000 bags) sample was examined and extensive measures were taken for bacterial culturing. While some units were heavily contaminated after just 24 hours of storage, the incidence as a whole varied according to the age of the concentrate and increased with the widespread practice of pooling individual units; over 30% of pools were contaminated at 3 days. See also D. H. Buccholz et al., New Eng. J. Med. 285:429 (1971). While other clinicians suggest lower numbers, recent studies indicate that septic platelet transfusions are significantly underreported. See e.g., J. F. Morrow et al., JAMA 266:555 (1991).
Pre-culturing platelets is not a solution to the bacterial contamination problem. The culture assay takes 48 hours to detect growth. Holding platelet units for an additional two days to await the results of the assay would create, ironically, a smaller margin of safety. See Table 2 in J. F. Morrow et al., JAMA 266:555 (1991). While heavily contaminated units would be detected at the outset, lightly contaminated units would be allowed to grow for two days. Older and potentially more contaminated units would end up being transfused.
Washing the blood cells (e.g., with saline) or filtering the bacteria are also not practical solutions. These techniques are time consuming and inefficient, as they can reduce the number of viable blood cells available for transfusion. Most importantly, they typically involve an "entry" into the storage system. Once an entry is made in a previously closed system, the system is considered "opened," and transfusion must occur quickly, regardless of the manner in which the blood was collected and processed in the first place.
Finally, antibiotics are not a reasonable solution. Contamination occurs from a wide spectrum of organisms. Antibiotics would be needed to cover this spectrum. Many recipients are allergic to antibiotics. In addition, there is an every increasing array of drug-resistant strains of bacteria that would not be inactivated.
In sum, there is a need for a means of inactivating organisms from blood components prior to storage and transfusion in a way that lends itself to use in a closed system. This approach must be able to handle a variety of organisms without harm to the blood product or the transfusion recipient.
The invention generally relates to synthetic media for use with blood preparations intended for in vivo use, including synthetic media used in conjunction with the photodecontamination of platelets. By the term "synthetic media" the present invention intends to indicate aqueous solutions (e.g., phosphate buffered, aqueous salt solutions) other than those found as natural fluids (e.g., plasma, serum, etc.). However, it is not intended that such synthetic media be used without the benefit of natural fluids. Indeed, in preferred embodiments, mixtures of synthetic salt solutions and natural fluids are contemplated.
The present invention contemplates that the activating means comprises a photoactivation device capable of emitting a given intensity of a spectrum of electromagnetic radiation comprising wavelengths between 180 nm and 400 nm, and in particular, between 320 nm and 380 nm. It is preferred that the intensity is less than 25 mW/cm12 (e.g. between 10 and 20 mW/cm2) and that the mixture is exposed to this intensity for between one and twenty minutes (e.g. ten minutes).
By the phrase "the maximum solubility of psoralen in water" the present invention intends a concentration derived experimentally in an aqueous solution in the absence of organic solvents (e.g., DMSO, ethanol, etc.) at approximately room temperature. Concentrations exceeding this level are detected by the presence of precipitate, which is undesirable for intravenous infusion.
A saturated solution of 8-methoxypsoralen can be made by simply dissolving the compound (over a number of hours at room temperature) in distilled water until precipitate is apparent. If the solution is simply centrifuged, the supernatant can have a concentration of over 50 μg/ml. On the other hand, if the solution is filtered (e.g., glass wool), the concentration of 8-methoxypsoralen has been found to be under 50 μg/ml. If, instead of centrifuging or filtering, the saturated solution is dialyzed against distilled water (over a number of days at room temperature), the compound is found to have a maximum solubility of approximately 39 μg/ml. It has been found that, when placed in a container (not glass) to shield the compound from the light, a 0.9% NaCl solution of 8-methoxypsoralen at a concentration of 30 μg/ml is stable.
When weighing components, some experimental variability is expected. The present invention employs the term "approximately" to reflect this variability. This variability is typically plus or minus 5% and usually less than 10%.
In one embodiment, the present invention contemplates a synthetic platelet storage media, comprising an aqueous solution of: 45-120 mM sodium chloride; 5-15 mM sodium citrate; 20-40 mM sodium acetate; and 20-30 mM sodium phosphate. In a preferred embodiment, the aqueous solution comprises: approximately 86 mM sodium chloride; approximately 10 mM sodium citrate; approximately 30 mM sodium acetate; and approximately 26 mM sodium phosphate. The solution has a pH of approximately pH 7.2 and an osmolarity of approximately 300mOsm/Kg. By not containing glucose or magnesium, the media is readily autoclaved.
FIG. 1 is a perspective view of one embodiment of the device of the present invention.
FIG. 2 is a cross-sectional view of the device shown in FIG. 1 along the lines of 2—2.
FIG. 3 is a cross-sectional view of the device shown in FIG. 1 along the lines of 3—3.
FIG. 4 is a cross-sectional view of the device shown in FIG. 1 along the lines of 4—4.
FIG. 5 schematically shows the decontamination approach of the present invention applied specifically to blood products.
FIG. 6 is a graph showing the photoaddition of 8-methoxypsoralen to nucleic acid.
FIG. 7 is a graph showing the degradation of 8-methoxypsoralen (8-MOP) compared to that of 4′-aminomethyl-4,5′,8-trimethylpsoralen (AMT), as measured by HPLC.
FIG. 8 is a graph showing the inactivation of gram negative bacteria.
FIG. 9A schematically shows the standard blood product separation approach used presently in blood banks.
FIG. 9B schematically shows an embodiment of the present invention whereby synthetic media is introduced to platelet concentrate prepared as in FIG. 9A.
FIG. 9C schematically shows one embodiment of the decontamination approach of the present invention applied specifically to platelet concentrate diluted with synthetic media as in FIG. 9B.
The invention generally relates to synthetic media for use with blood preparations intended for in vivo use, including synthetic media used in conjunction with the photodecontamination of platelets.
As noted previously, whole blood is collected and typically separated into red blood cells, platelets, and plasma. Each of these fractions are individually stored under specific conditions prior to in vivo use. In many cases, the extent of contamination is related to the storage time because of growth. A process that inactivated microorganisms at the time of blood collection would be expected to prevent growth during storage.
In one embodiment, the present invention contemplates inactivating blood products after separation but before storage. In this embodiment, a nucleic acid binding compound is selectively employed to treat contamination by microorganisms.
In one embodiment, the nucleic acid binding compound is selected from the group comprising furocoumarins. In a preferred embodiment, the furocoumarin is a psoralen that is activated by a photoactivation device.
Psoralens are tricyclic compounds formed by the linear fusion of a furan ring with a coumarin. Psoralens can intercalate between the base pairs of double-stranded nucleic acids, forming covalent adducts to pyrimidine bases upon absorption of longwave ultraviolet light (UVA). G. D. Cimino et al., Ann. Rev. Biochem. 54:1151 (1985); and Hearst et al., Quart. Rev. Biophys. 17:1 (1984). If there is a second pyrimidine adjacent to a psoralen-pyrimidine monoadduct and on the opposite strand, absorption of a second photon can lead to formation of a diadduct which functions as an interstrand crosslink. S. T. Isaacs et al., Biochemistry 16:1058 (1977); S. T. Isaacs et al., Trends in Photobiology (Plenum) pp. 279-294 (1982); J. Tessman et al., Biochem. 24:1669 (1985); and Hearst et al., U.S. Pat. Nos. 4,124,589, 4,169,204, and 4,196,281, hereby incorporated by reference.
Psoralens have been shown to inactivate viruses in some blood products. See H. J. Alter et al., The Lancet (ii:1446) (1988); L. Lin et al., Blood 74:517 (1989); and G. P. Wiesehahn et al., U.S. Pat. Nos. 4,727,027 and 4,748,120, hereby incorporated by reference, describe the use of a combination of 8-methoxypsoralen (8-MOP) and irradiation. They show that 300 μg/ml of 8-MOP together with one hour or more of irradiation with ultraviolet light can effectively inactivate viruses. However, these treatment conditions cause harm to the blood product because of energy transfer. Their approach is only feasible if the damage to cells is specifically suppressed by limiting the concentration of molecular oxygen, a difficult and expensive process.
The inactivation method of the present invention provides a method of inactivating single cell and multicellular organisms, and in particular, bacteria, fungi, mycoplasma and protozoa. In addition, the present invention contemplates inactivation of viruses (e.g. HIV virus). In contrast to previous approaches, the method of the present invention does not cause harm to the blood product. There is no significant damage to cells and, therefore, no need to limit the concentration of molecular oxygen.
The present invention contemplates using much lower concentrations of nucleic acid binding compounds than previously employed. For example, the present invention contemplates using 8-MOP at concentrations of 30 μg/ml or less. Indeed, a preferred concentration of 8-MOP for bacterial decontamination in platelet concentrates is 3 μg/ml or less, i.e., a one hundred-fold lower concentration than employed by G. P. Wiesehahn et al., supra. Because lower concentrations are employed, solvents like DMSO (used to increase the solubility of 8-MOP) are unnecessary.
The present invention, furthermore, contemplates using much lower doses of irradiation than previously described. This is accomplished with lower intensity irradiation sources, with wavelength cutoff filters (see below), and/or shorter irradiation times. In a preferred embodiment, the time of irradiation is variable and controlled from 1 second to 99 minutes, in one second increments.
In one embodiment, the device of the present invention is mounted on an agitator, giving horizontal unidirectional and sinusoidal motion of variable frequency and amplitude. In another embodiment, heat from the lamps, ballasts and other sources is blocked from the bags.
While it is not intended that the present invention be limited by the theory of inactivation, the use of lower compound concentrations and irradiation doses comes from an understanding that, where the present invention is applied to the decontamination of a single cell or multicellular organism, a lower level of nucleic acid binding will achieve inactivation. In addition, it is recognized that it is not essential that inactivation be complete. That is to say, partial inactivation will be adequate as long as the viable portion is unable, within the storage period, to grow to levels sufficient to cause disease.
To appreciate that, in any given case, an inactivation method may or may not achieve complete inactivation, it is useful to consider a specific example. A bacterial culture is said to be sterilized if an aliquot of the culture, when transferred to a fresh culture plate and permitted to grow, is undetectable after a certain time period. The time period and the growth conditions (e.g., temperature) define an "amplification factor". This amplification factor along with the limitations of the detection method (e.g., visual inspection of the culture plate for the appearance of a bacterial colony) define the sensitivity of the inactivation method. A minimal number of viable bacteria must be applied to the plate for a signal to be detectable. With the optimum detection method, this minimal number is 1 bacterial cell. With a suboptimal detection method, the minimal number of bacterial cells applied so that a signal is observed may be much greater than 1. The detection method determines a "threshold" below which the method appears to be completely effective (and above which the method is, in fact, only partially effective).
This interplay between the amplification factor of an assay and the threshold that the detection method defines, can be illustrated. For example, bacterial cells can be applied to a plate; the detection method is arbitrarily chosen to be visual inspection. Assume the growth conditions and time are such that an overall amplification of 104 has occurred. The detectable signal will be proportional to the number of bacterial cells actually present after amplification. For calculation purposes, the detection threshold is taken to be 106 cells; if fewer than 106 cells are present after amplification, no cell colonies are visually detectable and the inactivation method will appear effective. Given the amplification factor of 104 and a detection threshold of 106, the sensitivity limit would be 100 bacterial cells; if less than 100 viable bacterial cells were present in the original aliquot of the bacterial culture after the sterilization method is performed, the culture would still appear to be sterilized.
Such a situation is common for bacterial growth assays. The sensitivity of the assay is such that viable bacterial cells are present but the assay is unable to detect them. This may explain, at least in part, the variability in results obtained by researchers attempting to determine the extent of bacterial contamination of blood products. See D. H. Buchholz, et al., Transfusion 13:268 (1973), wherein such variability is discussed.
It should be noted that, in many countries, contamination of blood products by cellular organisms is more pervasive and, therefore, more serious than viral contamination. For example, in South America, the most important blood-borne organism is T. cruzi, which is the etiologic agent of Chagas disease. Approximately 16-18 million people are infected in the Americas (including 11% of the population of Chile). It is contemplated that the decontamination method of the present invention is well-suited for inactivation of this protozoa.
The present invention contemplates specific synthetic media formulations for use with blood preparations. The formulations of the present invention are particularly useful for platelet storage. These formulations are also useful when employed in conjunction with the photodecontamination of platelets.
While synthetic media have not previously been employed in conjunction with photodecontamination, synthetic media formulations have been described for platelet storage. For example, U.S. Pat. No. 4,828,976 to Murphy stresses the importance of having a synthetic blood platelet storage medium that is essentially free of glucose and calcium. The specification notes that glucose consumption and lactate production continue as long as glucose is present at the concentration above 1.5 mM (Col. 12, line 13). Lactate production is thought to be the cause of loss of pH stability and consequent impairment of platelet function. See also U.S. Pat. No. 4,925,665 and U.S. Pat. No. 4,992,363.
U.S. Pat. No. 4,447,415 to Rock describes a synthetic medium consisting essentially of: a balanced, physiologically compatible, saline solution; an anticoagulant; and one or more additives to enhance stability of the platelets selected from: (a) nutrients to improve the storage life of the platelets; (b) reversible inhibitors for platelet activation; (c) substances to raise cyclic adenosine monophosphate levels which have reversible effects on platelets; and (d) buffering agents which are physiologically compatible. The specification stresses the need to remove plasma. The specification indicates that this is achieved either by centrifugation and washing, or by "extraction" (see Examples 1 and 2 of the '415 patent). See also U.S. Pat. No. (Reissue) 32,874.
U.S. Pat. No. 4,704,352 to Miripol discloses a medium which contains either magnesium L-ascorbate-2-phosphate or calcium L-ascorbate-2-phosphate. U.S. Pat. No. 4,695,460 to Holme describe media that contain sodium bicarbonate. U.S. Pat. No. 4,390,619 to Harmening-Pittiglio discloses a media capable of supporting platelet metabolism, containing a water-insoluble polymer containing releasable phosphate or bicarbonate ions.
In contrast to these previous attempts, the formulations of the present invention: i) utilize residual plasma; and ii) avoid bicarbonate. With respect to plasma, the present invention contemplates that a standard plasma expression method (e.g., such as currently used in most blood banks) will be used, such that platelets are suspended in a residual plasma concentration between 8 and 25% by volume, and more commonly 12 to 20%. Bicarbonate has been found to be extremely difficult to control in a system employing gas permeable blood bags. The present invention therefore contemplates a phosphate buffer to control the pH of the blood preparation.
The present invention contemplates devices and methods for photoactivation and specifically, for activation of photoreactive nucleic acid binding compounds. The present invention contemplates devices having an inexpensive source of electromagnetic radiation that is integrated into a unit. In general, the present invention contemplates a photoactivation device for treating photoreactive compounds, comprising: a) means for providing appropriate wavelengths of electromagnetic radiation to cause activation of at least one photoreactive compound; b) means for supporting a plurality of blood products in a fixed relationship with the radiation providing means during activation; and c) means for maintaining the temperature of the blood products within a desired temperature range during activation. The present invention also contemplates methods, comprising: a) supporting a plurality of blood product containers, containing one or more photoreactive compounds, in a fixed relationship with a fluorescent source of electromagnetic radiation; b) irradiating the plurality of blood products simultaneously with said electromagnetic radiation to cause activation of at least one photoreactive compound; and c) maintaining the temperature of the blood products within a desired temperature range during activation.
The major features of one embodiment of the device of the present invention involve: A) an inexpensive source of ultraviolet radiation in a fixed relationship with the means for supporting the sample vessels; B) rapid photoactivation; C) large sample processing; D) temperature control of the irradiated samples; and E) inherent safety.
A. Electromagnetic Radiation Source
A preferred photoactivation device of the present invention has an inexpensive source of ultraviolet radiation in a fixed relationship with the means for supporting the sample vessels. Ultraviolet radiation is a form of energy that occupies a portion of the electromagnetic radiation spectrum (the electromagnetic radiation spectrum ranges from cosmic rays to radio waves). Ultraviolet radiation can come from many natural and artificial sources. Depending on the source of ultraviolet radiation, it may be accompanied by other (non-ultraviolet) types of electromagnetic radiation (e.g., visible light).
Particular types of ultraviolet radiation are herein described in terms of wavelength. Wavelength is herein described in terms of nanometers ("nm"; 10−9 meters). For purposes herein, ultraviolet radiation extends from approximately 180 nm to 400 nm. When a radiation source, by virtue of filters or other means, does not allow radiation below a particular wavelength (e.g., 320 nm), it is said to have a low end "cutoff" at that wavelength (e.g., "a wavelength cutoff at 300 nanometers"). Similarly, when a radiation source allows only radiation below a particular wavelength (e.g., 360 nm), it is said to have a high end "cutoff" at that wavelength (e.g., "a wavelength cutoff at 360 nanometers").
For any photochemical reaction it is desired to eliminate or least minimize any deleterious side reactions. Some of these side reactions can be caused by the excitation of endogenous chromophores that may be present during the photochemical activation procedure. In a system where only nucleic acid and psoralen are present, the endogenous chromophores are the nucleic acid bases themselves. Restricting the activation process to wavelengths greater than 320 nm minimizes direct nucleic acid damage since there is very little absorption by nucleic acids above 313 nm.
In blood products, the nucleic acid is typically present together with additional biological chromophores. If the biological fluid is just protein, the 320 nm cutoff will be adequate for minimizing side reactions (aromatic amino acids do not absorb above 320 nm). If the biological fluid includes cells and/or cellular constituents, there will be many other chromophores, including hemes and flavins.
Hemes are abundant in blood products where they arise from the lysis of red cells. Flavins, like hemes, are required for metabolic respiration. Both of these endogenous chromophores will cause damage to cells if excited by photoirradiation.
Hemes have three principle absorption bands: two are in the red region of the visible spectrum; the other is centered about 400 nm. Flavins have two principle absorption peaks: one at 450 nm and the other at 370 nm.
In view of the presence of these endogenous chromophores in blood products, it is intended that the device of the present invention be designed to allow for irradiation within a small range of specific and desirable wavelengths, and thus avoid damage to cells caused by energy transfer. The preferred range of desirable wavelengths is between 320 and 350 nm.
Some selectivity can be achieved by choice of commercial irradiation sources. For example, while typical fluorescent tubes emit wavelengths ranging from 300 nm to above 400 nm (with a broad peak centered around 360 nm), BLB type fluorescent lamps are designed to remove wavelengths above 400 nm. This, however, only provides an upper end cutoff.
In a preferred embodiment, the device of the present invention comprises an additional filtering means. In one embodiment, the filtering means comprises a glass cut-off filter, such as a piece of Cobalt glass. In another embodiment, the filtering means comprises a liquid filter solution that transmit only a specific region of the electromagnetic spectrum, such as an aqueous solution of Co(No3)2. This salt solution yields a transmission window of 320-400 nm. In a preferred embodiment, the aqueous solution of Co(No3)2 is used in combination with NiSO4 to remove the 365 nm component of the emission spectrum of the fluorescent or arc source employed. The Co-Ni solution preserves its initial transmission remarkably well even after tens of hours of exposure to the direct light of high energy sources.
It is not intended that the present invention be limited by the particular filter employed. Several inorganic salts and glasses satisfy the necessary requirements. For example, cupric sulfate is a most useful general filter for removing the infra-red, when only the ultraviolet is to be isolated. Its stability in intense sources is quite good. Other salts are known to one skilled in the art. Aperture or reflector lamps may also be used to achieve specific wavelengths and intensities.
When ultraviolet radiation is herein described in terms of irradiance, it is expressed in terms of intensity flux (milliwatts per square centimeter or "mW cm−2"). "Output" is herein defined to encompass both the emission of radiation (yes or no; on or off) as well as the level of irradiance. In a preferred embodiment, intensity is monitored at 4 locations: 2 for each-side of the plane of irradiation.
A preferred source of ultraviolet radiation is a fluorescent source. Fluorescence is a special case of luminescence. Luminescence involves the absorption of electromagnetic radiation by a substance and the conversion of the energy into radiation of a different wavelength. With fluorescence, the substance that is excited by the electromagnetic radiation returns to its ground state by emitting a quantum of electromagnetic radiation. While fluorescent sources have heretofore been thought to be of too low intensity to be useful for photoactivation, in one embodiment the present invention employs fluorescent sources to achieve results thus far achievable on only expensive equipment.
As used here, fixed relationship is defined as comprising a fixed distance and geometry between the sample and the light source during the sample irradiation. Distance relates to the distance between the source and the sample as it is supported. It is known that light intensity from a point source is inversely related to the square of the distance from the point source. Thus, small changes in the distance from the source can have a drastic impact on intensity. Since changes in intensity can impact photoactivation results, changes in distance are avoided in the devices of the present invention. This provides reproducibility and repeatability.
Geometry relates to the positioning of the light source. For example, it can be imagined that light sources could be placed around the sample holder in many ways (on the sides, on the bottom, in a circle, etc.). The geometry used in a preferred embodiment of the present invention allows for uniform light exposure of appropriate intensity for rapid photoactivation. The geometry of a preferred device of the present invention involves multiple sources of linear lamps as opposed to single point sources. In addition, there are several reflective surfaces and several absorptive surfaces. Because of this complicated geometry, changes in the location or number of the lamps relative to the position of the samples to be irradiated are to be avoided in that such changes will result in intensity changes.
B. Rapid Photoactivation
The light source of the preferred embodiment of the present invention allows for rapid photoactivation. The intensity characteristics of the irradiation device have been selected to be convenient with the anticipation that many sets of multiple samples may need to be processed. With this anticipation, a fifteen minute exposure time or less is a practical goal.
In designing the devices of the present invention, relative position of the elements of the preferred device have been optimized to allow for fifteen minutes of irradiation time, so that, when measured for the wavelengths between 320 and 350 nanometers, an intensity flux greater than approximately 1 mW cm−2 is provided to the sample vessels. In a preferred embodiment, the device irradiates both sides of the bag.
C. Processing of Large Numbers of Samples
As noted, another important feature of the photoactivation devices of the present invention is that they provide for the processing of large numbers of samples. In this regard, one element of the devices of the present invention is a means for supporting a plurality of blood products, and in particular, blood bags. In the preferred embodiment of the present invention the supporting means comprises glass plates between two banks of lights with a capacity of six 50 ml bags (equivalent to Dupont Stericell bag) plus connectors and tubing, at one time. By accepting commonly used commercially available blood bags, the device of the present invention allows for convenient processing of large numbers of samples.
D. Temperature Control
As noted, one of the important features of the photoactivation devices of the present invention is temperature control. Temperature control is important because the temperature of the sample in the sample at the time of exposure to light can dramatically impact the results. For example, conditions that promote secondary structure in nucleic acids also enhance the affinity constants of many psoralen derivatives for nucleic acids. Hyde and Hearst, Biochemistry, 17, 1251 (1978). These conditions are a mix of both solvent composition and temperature. With single stranded 5S ribosomal RNA, irradiation at low temperatures enhances the covalent addition of HMT to 5S rRNA by two fold at 4� C. compared to 20� C. Thompson et al., J. Mol. Biol. 147:417 (1981). Even further temperature induced enhancements of psoralen binding have been reported with synthetic polynucleotides. Thompson et al., Biochemistry 21:1363 (1982).
With respect to bacteria, it should be noted that repair of crosslinks occurs during irradiation. However, where a lower temperature is employed during irradiation, the bacterial repair process is suppressed. Thus, a 15� C. irradiation has a significant effect on the level of inactivation that is observed.
E. Inherent Safety
Ultraviolet radiation can cause severe burns. Depending on the nature of the exposure, it may also be carcinogenic. The light source of a preferred embodiment of the present invention is shielded from the user. This is in contrast to the commercial hand-held ultraviolet sources as well as the large, high intensity sources. In a preferred embodiment, the irradiation source is contained within a housing made of material that obstructs the transmission of radiant energy (i.e., an opaque housing). No irradiation is allowed to pass to the user. This allows for inherent safety for the user.
In the experimental disclosure which follows, the following abbreviations apply: eq (equivalents) M (Molar); μM (micromolar); N (Normal); mol (moles); mmol (millimoles); μmol (micromoles); nmol (nanomoles); gm (grams); mg (milligrams); μg (micrograms); L (liters); ml (milliliters); μl (microliters); cm (centimeters); mm (millimeters); μm (micrometers); nm (nanometers); � C. (degrees Centigrade); HPLC (High Pressure Liquid Chromatography).
In some of the examples below, phosphate buffered synthetic media is formulated for platelet storage and treatment. One embodiment is made as follows:
Distilled water is added to make 1 liter, the solution is mixed, sterile filtered (0.2 micron filter) and refrigerated.
The following examples serve to illustrate certain preferred embodiments and aspects of the present invention and are not to be construed as limiting the scope thereof.
As noted above, the present invention contemplates devices and methods for the activation of photoreactive nucleic acid binding compounds. In this example, a photoactivation device is described for decontaminating blood products according to the method of the present invention. This device comprises: a) means for providing appropriate wavelengths of electromagnetic radiation to cause activation of at least one photoreactive compound; b) means for supporting a plurality of blood products in a fixed relationship with the radiation providing means during activation; and c) means for maintaining the temperature of the blood products within a desired temperature range during activation.
FIG. 1 is a perspective view of one embodiment of the device integrating the above-named features. The figure shows an opaque housing (100) with a portion of it removed, containing an array of bulbs (101) above and below a plurality of representative blood product containing means (102) placed between plate assemblies (103, 104). The plate assemblies (103, 104) are described more fully, subsequently.
The bulbs (101), which are connectable to a power source (not shown), serve as a source of electromagnetic radiation. While not limited to the particular bulb type, the embodiment is configured to accept an industry standard, dual bipin lamp.
The housing (101) can be opened via a latch (105) so that the blood product can be placed appropriately. As shown in FIG. 1, the housing (100), when closed, completely contains the irradiation from the bulbs (101). During irradiation, the user can confirm that the device is operating by looking through a safety viewport (106) which does not allow transmission of ultraviolet light to the user.
The housing (100) also serves as a mount for several electronic components on a control board (107), including, by way of example, a main power switch, a count down timer, and an hour meter. For convenience, the power switch can be wired to the count down timer which in turn is wired in parallel to an hour meter and to the source of the electromagnetic radiation. The count down timer permits a user to preset the irradiation time to a desired level of exposure. The hour meter maintains a record of the total number of radiation hours that are provided by the source of electromagnetic radiation. This feature permits the bulbs (101) to be monitored and changed before their output diminishes below a minimum level necessary for rapid photoactivation.
FIG. 2 is a cross-sectional view of the device shown in FIG. 1 along the lines of 2—2. FIG. 2 shows the arrangement of the bulbs (101) with the housing (100) opened. A reflector (108A, 108B) completely surrounds each array of bulbs (101). Blood product containing means (102) are placed between upper (103) and lower (104) plate assemblies. Each plate assembly is comprised of an upper (103A, 104A) and lower (103B, 104B) plates. The plate assemblies (103, 104) are connected via a hinge (109) which is designed to accommodate the space created by the blood product containing means (102). The upper plate assembly (103) is brought to rest gently on top of the blood product containing means (102) supported by the lower plate (104B) of the lower plate assembly (104).
Detectors (110A, 11OB, 11OC, 11OD) may be conveniently placed between the plates (103A, 103B, 104A, 104B) of the plate assemblies (103, 104). They can be wired to a printed circuit board (111) which in turn is wired to the control board (107).
FIG. 3 is a cross-sectional view of the device shown in FIG. 1 along the lines of 3—3. Six blood product containing means (102) (e.g., Teflon™ platelet unit bags) are placed in a fix relationship above an array of bulbs (101). The temperature of the blood product can be controlled via a fan (112) alone or, more preferably, by employing a heat exchanger (113) having cooling inlet (114) and outlet (115) ports connected to a cooling source (not shown).
FIG. 4 is a cross-sectional view of the device shown in FIG. 1 along the lines of 4—4. FIG. 4 more clearly shows the temperature control approach of a preferred embodiment of the device. Upper plate assembly plates (103A, 103B) and lower plate assembly plates (104A, 104B) each create a temperature control chamber (103C, 104C), respectively. The fan (112) can circulate air within and between the chambers (103C, 104C). When the heat exchanger (113) is employed, the circulating air is cooled and passed between the plates (103A, 103B, 104A, 104B).
FIG. 5 shows an embodiment wherein platelets are treated by the method of the present invention. Following fractionation, platelets are transferred to a bag containing a nucleic acid binding compound (shown in FIG. 5 as a shaded bag). This bag, which has transmission properties and other characteristics suited for the present invention, is then placed in an irradiation device (such as that described in Example 1, above) and is irradiated. The free compound may be collected or "captured" as desired by a capture device. In such a case, the bag would contain only compound that is contained in cells; the bag would have no free compound (this bag is indicated in FIG. 5 as unshaded).
In one capture approach, a bag comprised of a psoralen-binding polymer is employed to capture the compound. The Cutter CLX bag has been found to have this property.
In this example, the decontamination methods of the present invention are applied to inactivate Klebsiella pneumoniae, which is known to be among the organisms associated with bacteremia. See generally, Infect. Control 5:343 (1984). The particular isolate in this case was obtained following a platelet transfusion where the recipient immediately went into shock and later died. The platelet bag was obtained and cultured, and the organism was identified and serotyped.
For the experiment, the strain was kept at ambient temperature and inoculated onto either heart infusion agar (HIA) or heart infusion agar containing 5% (v/v) sheep blood (BAP) by swabbing each plate for confluency via a sterile applicator swab. Cultures were then incubated under static conditions for 16-18 h at 35� C. At the end of the incubation period, cultures were removed and suspended in phosphate buffered saline (PBS;pH 7.2-7.4) and spectrophotometrically standardized to 1.0 at an OD610 using a Spectronic 501 or 601 spectrophotometer (Bausch and Lomb). After standardization, suspensions were diluted 1:10 in PBS to achieve an ca. 108 CFU/ml concentration. This standardized suspension is then split to use an aliquot for the inactivation study, while another portion was plated in duplicate 10-fold serial dilutions onto HIA (or BAP) to ensure appropriate concentrations of the organism.
To assess inactivation of the organism, two ABO compatible freshly outdated human platelet concentrate units were obtained from the Blood Bank of Alameda—Contra Costa Medical Association. They were pooled and redivided into two bags. One bag was infused with the bacteria preparation. The platelets in the second unit were pelleted at 4000 �g for 6 minutes and then resuspended in a medium containing 85% saline and 15% plasma. Bacteria was added after platelets were well resuspended.
3 ml aliquots of bacteria containing platelet concentrate were transferred to a Teflon™ minibag (American Fluoroseal Corporation, Silver Spring, Md.) and received specified amounts of 8-MOP and UVA irradiation, except for the controls, which were irradiated without psoralen, or received no treatment. Temperature was maintained at 25� C. during irradiation in the irradiation device described above which is equipped with an air cooling mechanism.
F24T 12-BL-HO fluorescent lamps were used. These are high output "black light" tubes (engineered to emit specific wavelengths by means of an internal phosphor coating) 24 inches in length. Total intensity is less than 25 mw/cm2 and typically between 15 and 20 mw/cm2.
Following irradiation, bacteria were quantified by plating 0.1 ml of serial 10-fold dilutions in broth onto 100 mm petri dishes containing BHI agar. After 24 hr incubation at 35� C., colonies were counted and bacterial concentration was calculated on a per ml basis. The results (FIG. 8) show that while only 1.2 logs of Klebsiella were killed by 5 μg/ml 8-MOP in five minutes of UVA irradiation in 100% plasma, more than 6.5 logs were killed under the same conditions in 15% plasma and 85% saline.
While not limited to any theory as to the mechanism by which this improvement in decontamination efficiency was achieved, it would appear that these results indicate that either that the optical properties of the synthetic medium are better, or the lower protein concentration allows for a higher concentration of free 8-MOP. In the latter situation, the 8-MOP would be more available for decontamination.
Artuc and co-workers examined the solubility of 8-MOP in human and bovine serum proteins. They showed that, when working with 8-MOP concentrations ranging from 100 to 1000 ng/ml in serum, 75% to 80% of the 8-MOP was bound to albumin. M. Artuc et al., Brit. J. Derm. 101:669 (1979).
In this example, the binding of 8-MOP to Calf Thymus DNA is compared using plasma and a protein free media in order to validate the efficiency of psoralen-nucleic interactions under the decontamination methods of the present invention. Although this measurement used eukaryotic nucleic acid rather than bacterial nucleic acid, it is a useful indicator of the degree of adduct formation for bacteria.
3H-8-MOP was prepared to a concentration of 115 μg/ml in ethanol at a specific activity of 4.7�106 CPM/microgram (hereinafter "8-MOP stock"). Thereafter 130.5 or 22 μl of 8-MOP stock (2 each) for samples containing DNA ("+DNA") and 52.2 or 8.7 μl for samples not containing DNA ("−DNA") were dried down. To+DNA samples, 40 μl of DNA stock (7.7 mg/ml) was added as well as either 460 μl plasma (day old frozen) or 450 μl Tris-EDTA ("TE") buffer. To the latter was also added 10 μl 5M NaCl. For—DNA samples (i.e., the controls), 184 μl plasma and 16 μl water was added.
The samples were mildly vortexed for approximately one hour and the counts were checked to confirm that the 8-MOP is dissolved.
Each sample (100 μl) was irradiated on an HRI-100 (HRI Research Inc., Concord, Calif.) at 25� C. for 0, 2, 4, 8, and 16 minutes. Samples were kept at 4� C. overnight after irradiation. Thereafter, the samples were extracted. First, a phenol solution was prepared at pH 8 by equilibrating with 0.1 M Tris pH 8. Each sample was then extracted with 100 μl phenol. Each sample was centrifuged for 5 minutes to remove the aqueous phase to a new tube. A second extraction was performed with 100 μl 1:1 phenol:chloroform. A final extraction was performed with 100 μl chloroform.
The final aqueous phase was precipitated by adding 50 μl NaCl adjusted to give a final concentration of NaCl of 0.2 M and then adding 250 μl ethanol. The samples were again centrifuged (10 minutes). The supernatant was removed and the pellets were dried. The pellets were resuspended in 100 μl TE and reprecipitated. This was repeated for a total of 3 precipitations. The final pellets were brought up in 600 μl water and 100 μl was counted. Each sample was assayed for DNA by measuring absorbance (260 nm). 8-MOP levels were plotted as adducts per 1000 base pairs ("8-MOP:kBP").
The results (FIG. 6) show that plasma does significantly change the addition kinetics of 8-MOP to DNA. Addition to nucleic acid is much better in the protein free media.
The frequency of 8-MOP-DNA adduct formation in protein free media predicts a high multiplicity of modification of the bacterial genome. Furthermore, this type of biochemical measurement has the potential to provide a means to monitor the efficiency of the photochemical inactivation method.
Photoactivation of psoralens and isopsoralens may result in a variety of photoproducts. "Photoproduct" is best understood by considering the possible reactions of photoreactive compound when exposed to activating wavelengths of electromagnetic radiation. While not limited to any precise mechanism, it is believed that the reaction of photoreactive compound in its ground state ("C") with activating wavelengths of electromagnetic radiation creates a short-lived excited species ("C*"):
What happens next is largely a function of what potential reactants are available to the excited species. Since it is short-lived, a reaction of this species with nucleic acid ("NA") is believed to only be possible if nucleic acid is present at the time the excited species is generated. Thus, the reaction must, in operational terms, be in the presence of activating wavelengths of electromagnetic radiation, i.e., it is "photobinding"; it is not dark binding. The reaction can be depicted as follows:
The product of this reaction is hereinafter referred to as "Photoaddition Product" and is to be distinguished from "Photoproduct."
With this reaction described, one can now consider the situation where nucleic acid is not available for binding at the time the compound is exposed to activating wavelengths of electromagnetic radiation. Since the excited species is short-lived and has no nucleic acid to react with, the excited species may simply return to its ground state:
On the other hand, the excited species may react with itself (i.e., a ground state or excited species) to create a ground state complex ("C:C"). The product of these self-reactions where two compounds react is referred to as "photodimer" or simply "dimer." The self-reactions, however, are not limited to two compounds; a variety of multimers may be formed (trimers, etc.).
The excited species is not limited to reacting with itself. It may react with its environment, such as elements of the solvent ("E") (e.g., ions, gases, etc.) to produce other products:
It is this type of reaction that is believed to cause cellular damage (e.g., reaction with oxygen to create singlet oxygen species). Furthermore, it may simply internally rearrange ("isomerize") to a ground state derivative ("["):
Finally, the excited species may undergo other reactions than described here.
The present invention and the understanding of "photoproduct" does not depend on which one (if any) of these reactions actually occurs. "Photoproduct"—whatever its nature—is deemed to exist if, following the reaction of a compound and activating wavelengths of electromagnetic radiation, there is a resultant product formed that can interact with other components of the reaction environment.
With psoralens such as 4′-hydroxymethyl-4,5′,8-tri-methylpsoralen (HMT), there are a number of resultant products produced when the HMT is exposed to activating wavelengths of electromagnetic radiation. The major resultant products of HMT are two cyclobutyl photodimers. In one of the diners, the two pyrone rings are linked in a cis-syn configuration, while in the other dimer, the linkage occurs between the furan end of one molecule and the pyrone end of the other, again with cis-syn configuration. A third resultant product of HMT is a monomeric HMT photoisomer. In this isomer, the central ring oxygens assume a 1, 4 instead of the normal 1, 3 orientation. While the two photodimers would not be expected to have an intercalating activity due to geometrical considerations, the photoisomer remains planar, and accordingly, it is contemplated that it has a positive intercalative association with double stranded nucleic acid and, thus, could be a mutagen.
In this example, the photochemical breakdown of 8-MOP is compared with AMT. The samples were analyzed by reverse phase HPLC using a Rainen Dynamax 300A column. Gradient elution was performed with 0.1 M ammonium acetate / acetonitrile (0-70% acetonitrile over 42 minutes). AMT elutes as a single peak at approximately 24 minutes under these conditions. Detection was by absorption at either 260 or 330 nm. The latter wavelength was used for the plasma containing samples.
Standard solutions of each compound were prepared at various concentrations. These solutions were then diluted 1:10 into water, then 300 μl injected for analysis. All samples were monitored at 300 nm. Peaks were analyzed by measuring either peak height or peak area, then converted to a μh/ml value using the standard plot. Peak area was determined by photocopying the trace, cutting out the copy of the peak, then weighing the resultant trace. The two methods gave essentially the same result.
The results are shown in FIG. 7. Clearly, AMT degrades more quickly than 8-MOP. It would, therefore, be expected to generate more photoproducts—which eventually would end up in the transfusion recipient. By contrast, it is not expected that 8-MOP generates a significant amount of photoproducts. This is important when one considers that the weight of authority has concluded that unactivated 8-MOP is nonmutagenic.
FIG. 9 shows an embodiment wherein platelets are diluted with synthetic media by the method of the present invention. FIG. 9A schematically shows the standard blood product separation approach used presently in blood banks. Three bags are integrated by flexible tubing to create a blood transfer set (200) (e.g., commercially available from Baxter, Deerfield, Ill.). After blood is drawn into the first bag (201), the entire set is processed by centrifugation (e.g., Sorvall™ swing bucket centrifuge, Dupont), resulting in packed red cells and platelet rich plasma in the first bag (201). The plasma is expressed off of the first bag (201) (e.g., using a Fenwall™ device for plasma expression), through the tubing and into the second bag (202). The first bag (201) is then detached and the two bag set is centrifuged to create platelet concentrate and platelet-poor plasma; the latter is expressed off of the second bag (202) into the third bag (203).
FIG. 9B schematically shows an embodiment of the present invention by which synthetic media is introduced to platelet concentrate prepared as in FIG. 9A. A two bag set (300) is sterile docked with the platelet concentrate bag (202) (indicated as "P.C."). Sterile docking is well-known to the art. See e.g., U.S. Pat. No. 4,412,835 to D. W. C. Spencer, hereby incorporated by reference. See also U.S. Pat. Nos. 4,157,723 and 4,265,280, hereby incorporated by reference. Sterile docking devices are commercially available (e.g., Terumo, Japan).
One of the bags (301) of the two bag set (300) contains a synthetic media formulation of the present invention (indicated as "STERILYTE"). In the second step shown in FIG. 9B, the platelet concentrate is mixed with the synthetic media by transferring the platelet concentrate to the synthetic media bag (301).
FIG. 9C schematically shows one embodiment of the decontamination approach of the present invention applied specifically to platelet concentrate diluted with synthetic media as in FIG. 9B. In this embodiment, platelets have been transferred to a synthetic media bag (301) containing a nucleic acid binding compound. This bag (301), which has UV light transmission properties and other characteristics suited for the present invention, is then placed in a device (such as that described in Example 1, above) and illuminated.
Following phototreatment, the platelets are transferred from the synthetic media bag (301) into the storage bag (302) of the two bag set (300). The storage bag can be a commercially available storage bag (e.g., CLX bag from Cutter).
From the above, it should be evident that the present invention provides synthetic media formulations for use with blood preparations intended for storage and in vivo use. The formulations include synthetic media formulations to be employed in conjunction with the photodecontamination of platelets.
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This job is no longer available
Further information
Our client is a leading international property and construction developer with a reputation for being involved in key land mark projects throughout the world covering major real estate for blue chip portfolios of clients. They wish to appoint a highly experience Project Director for a major new development.
The Role
- Reporting into the UK and Ireland Operations Director you will hold full programme accountability managing key project milestones and commercial returns are protected.
- In a highly client facing role you will ensure client and partner expectations are met and the programme is delivered to the appropriate quality levels.
The Requirement
- Degree qualified you will have a proven track record of delivering major land mark projects with a blue chip international developer.
- You will possess the necessary communication skills and gravitas to lead multidisciplined teams with strong commercial acumen.
In the first instance please forward relevant CV's by email including your current remuneration package quoting PD/159/25


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text:
This is a look I did using Chanel Illusion D'Ombre Long Wear Luminous Eyeshadow in #84 Epatant. It's the first time that I used this eyeshadow since purchasing it. And not only was I so impressed with the quality of this product but the shade is so beautiful yet gorgeous that it even drew the attention and garnered praises from the SAs of Benefit and Urban Decay!
*I had drafted the first paragraph last month and slap myself for not finishing up this post, though the photos have been cropped!
Face Of The Day
MAC Prep+Prime Skin Base mixed with Juice Beauty Perfecting Foundation in Organic Sand
Guerlain Meteorites in Mythic
The Body Shop Roseflower Blush
Make Up For Ever Fascinating lip gloss #16
Eye Of The Day
Chanel Illusion D'Ombre Long Wear Luminous Eyeshadow in #84 Epatant all over lids and blend upward all the way to almost the brow.
Finish off with MAC Zoom Lash Zoomblack mascara.
More photos............
I think I will be using this eyeshadow a lot come the Holidays, wearing it to dinners and parties! :D
Happy Monday!


text:
One of the foremost standard haircut these days is that the bob cut. this can be as a result of the very fact that it doesn't solely cause you to look nice or that your hair can look longer than it really is, however additionally as a result of you may not ought to manage any kind of bother whereas styling it. once it involves ladies, it's extremely exhausting to form everything work well along. we have a tendency to area unit talking concerning make-up, clothes, jewellery and hair. But, once you have a bob haircut, the opposite things simply appear to feature up and switch the total image into a reasonably superb one. It simply depends on you what kind of bob haircut would you favor for yourself. There area unit all kinds of variations that you simply might make a choice from and therefore the look that you simply can have can depend upon the length of your hair, the color and the way dramatic would you favor it. Here, you may realize a range of the foremost superb bob haircuts that you simply will select from! Short Hair Styles For Women 2014
Regular Bob Hair
This is clearly one in all the standard kinds of bob that you simply might see on the road. However, you must understand that it's that standard as a result of the very fact that it extremely will amendment the approach you look. it's gorgeous!


text:
Faced with a shocking case of a teacher accused of playing classroom sex games with children for years, Los Angeles schools Superintendent John Deasy delivered another jolt: He removed the school's entire staff — from custodians to the principal — to smash what he called a "culture of silence."
"It was a quick, responsible, responsive action to a heinous situation," he said. "We're not going to spend a long time debating student safety."
The controversial decision underscores the 51-year-old superintendent's shake-up of the lethargic bureaucracy at the nation's second-largest school district. His swift, bold moves have rankled some and won praise from others during his first year of leadership.
Hired with a mandate to boost achievement in the 660,000-pupil Los Angeles Unified School District, Deasy has become known for 18-hour days that involve everything from surprise classroom visits and picking up playground litter to lobbying city elite for donations and blasting Sacramento politicians over funding cuts.


text:
UGA WR Malcolm Mitchell Injures Knee
Malcolm Mitchell
Aug. 1, 2014
ATHENS -- University of Georgia junior wide receiver Malcolm Mitchell suffered a right knee injury earlier this week while running routes with teammates.
He underwent an arthroscopic procedure Thursday to address a cartilage injury. A full recovery is anticipated; however, no timetable is set for his return at this point according to UGA Sr. Associate Athletic Director and Director of Sports Medicine Ron Courson.


text:
- CrackBerry Addict
11-09-12, 03:44 AM Thread Author #1
- Posts
- 656 Posts
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The BlackBerry 10 homescreen on the PlayBook
Will the BlackBerry 10 homescreen work in Landscape? If they are planning on BB10 for the PlayBook.. it needs to, since PlayBook is based on Landscape use.
Also, how many active frames would RIM allow the PlayBook to have open? The PlayBook's screen is larger but, it's a similar resolution. The processor is also dated, and lack of memory and RAM would not allow the OS to 'flow' as it will on the phones.
Personally I hope RIM releases a updated WiFi PlayBook that is based for portrait use, with a newer processor, 2GB RAM, NFC, and an SD card slot since the BlackBerry 10 phones are getting expandable memory storage.
11-09-12, 09:29 AM #2Forum Rules: Community Rules & Guidelines Quick Help: BLACKBERRY HELP 101 | OS Arquive: BB10 | Direct load Android (APK) Apps : BB10 (10.2.1+) |Sideloading : BB10 10.2.0-/PlayBook |Google Play Store: BB10 |Update/Reload Software: BBOS - PlayBook - BB10 | Debrick: BBOS - PlayBook - BB10 | Backup 3rd Party Apps: BBOS | Battery Saving : BBOS - PlayBook - BB10 | Dead Battery: Stack Charging | BB AppWorld: Refunds
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My Apps: Papo GP | My Channel: Project Management
- CrackBerry Abuser
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Re: The BlackBerry 10 homescreen on the PlayBook
Nobody knows what it looks like.. Unless Kevin, Blaze or Adam have had a secret peek at it in RIM internals
- CrackBerry Addict
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- 543 Posts
Re: The BlackBerry 10 homescreen on the PlayBook
The only thing I would add to the above responses is that the PlayBook should still work in both portrait and landscape. All phones and tablets have both orientations for the OS but apps designers can choose to use one or both. If anything, RIM will try to provide both orientations more often than just landscape only. Personally, I will be happy to see some more of the landscape only apps have both portrait and landscape mode.
- CrackBerry Abuser
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Re: The BlackBerry 10 homescreen on the PlayBook
It is important that BB10 is Optimized for Landscape AND Portrait orientations, Modern day smartphones and tablets cannot be tied down to one orientation, and have spotty support for the other, ...
The Playbook manages both orientations flawlessly, but we have yet to see landscape anything from bb10, it worries me.
- CrackBerry Abuser
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Re: The BlackBerry 10 homescreen on the PlayBook
my guess bb10 will do both modes just fine, maybe there is no landscape homescreen on the phone version but the tablet version will have it for sure.
RIM has to deliver a OS with system apps that work in both ways, anything else would just be ridiculous in 2013....
- CrackBerry Genius of Geniuses
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Re: The BlackBerry 10 homescreen on the PlayBook
One of the requirements for our apps to get the Built For BlackBerry certification will be properly supporting not only the physical keyboard phone models, but also the landscape/portrait change. In some instances this of course won't make sense (e.g. some games) but in cases where it would, it appears it will be a mandatory feature.
I think that also makes it pretty certain that the system apps will generally support landscape mode as well, since they'd look pretty silly forcing developers to do that, yet not do it themselves.Battery Guru for BB10 tracks voltage, power, battery health. (Also on the PlayBook.) White Noise helps you sleep or concentrate.
Follow us on Twitter or Facebook or visit Engenuity's mobile apps page.Thanked by 3:
BerryClever (11-13-12), Dr.Deebs (11-12-12), kbz1960 (11-12-12)
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text:
Brooklyn Stoop
Words by Rachel Jones Photograph by Rachel Jones & Adam Patrick Jones
A couple gets to know the neighbors on a Brooklyn stoop, which has become their makeshift outdoor community living room.
Before I moved to New York, I didn't know what a stoop was. My introduction came recently, when my boyfriend moved into a renovated pre-war brownstone in Brooklyn. I walked up the steps, surveyed the meager shreds of green still alive within a jumble of pots, and hurriedly closed the door behind me.
Several weeks later, his roommate organized a stoop party on the steps. We joined in, cautiously. Somehow, the idea of getting to know the rest of the building's residents seemed foreign. Up until that moment, I'd lived in the same apartment for two years without ever crossing paths with my upstairs neighbor.
The party, small affair that it was, marked a clear before and after. Here's why. When you live in a place where space is viewed as a commodity, a luxury even, you become greedy about it. You clutch it close, and maybe only invite in those dearest. Stoop life challenges that notion. You are living in the open. Without a hedge to block out inquisitive eyes. Without a wall to buffer the whine of the ice-cream truck. In the simplest of ways, you are forced to relearn the art of sharing.
Over time, the stoop has become our outdoor living room. Those stains, scuff marks, and odds and ends that created the backstory of the building now feed into my story. The Italian espresso can, a makeshift cigarette holder, that has prompted many a smoke break and an excuse for a beer on the steps. The assortment of gardening tools jumbled in the entryway corner that heralds the beautification process of the stoop's plant life. The spatter of dark spots on the third step, left over from a particularly delicious slice of Neapolitan-style pizza. The small plastic water bowl, hidden behind a planter, that signals that this stoop has been adopted by a neighborhood Siamese, still unnamed.
The trail of crumbs, both figurative and real, has become clues about the comings and goings of the residents at 371A. They are frequent patrons of the local taqueria. A green thumb in the building is sorely needed. As is a broom. But mostly, they are happy and easygoing. They are lucky to have each other as neighbors.
I don't even live in the building, but I see how the stoop has brought them together. Somehow, it changes things. Just like it did for the cat.


text:
On February 5, 2008, the closing price of the Company's common stock was
$3.81 per share
IRVINE, Calif., Feb. 5 /PRNewswire-FirstCall/ -- IDM Pharma, Inc. (Nasdaq: IDMI) today announced that it received a letter, dated February 4, 2008, from The NASDAQ Stock Market, notifying IDM Pharma that during the preceding 30 consecutive business days, the closing bid price of the Company's common stock had been below the $1.00 minimum bid per share required for continued listing on the NASDAQ Global Market under NASDAQ Marketplace Rule 4450(a)(5).
The NASDAQ letter dated February 4, 2008 has no immediate effect on the listing of the Company's common stock. On February 5, 2008, the closing price of the Company's common stock was $3.81 per share.
In accordance with NASDAQ Marketplace Rule 4450(e)(2), the Company will
be granted 180 calendar days, or until August 4, 2008, to regain compliance
with the minimum bid price requirement in accordance with NASDAQ
Marketplace Rule 4450(a)(5) by maintaining a closing bid price of $1.00 per
share or higher for a minimum of 10 consecutive trading days. If the
Company is successful in meeting this requirement, NASDAQ will provide
written notification that is has achieved compliance with the Rule. If the
Company is unsuccessful in meeting the minimum bid requirement during this
initial 180-day period, NASDAQ will provide notice to IDM Pharma that the
Company's common stock will be delisted from the NASDAQ Global Market. If
the Company receives a delisting notification, the Company may appeal to
the Listing Qualifications Panel or apply to transfer its common stock to
the NASDAQ Capital Market if IDM Pharma satisfies all criteria for initial
listing on the NASDAQ C
|SOURCE IDM Pharma, Inc.
Copyright©2008 PR Newswire.
All rights reserved


text:
Apr 26 2012
Learning American Sign Language
In one more week Emelie will complete her second full year of American Sign Language (ASL). During her first year, ASL I, Emelie learned approximately 600 signs over the course of the year and how to carry on everyday conversations with a Deaf person–including an understanding of basic ASL grammatical rules (facial expressions, body language, body shifting/direct address, listing, eye gaze, classifiers and gesturing). Emelie's teacher also worked with the class to develop 'sign to voice' interpreting skills and an awareness of Deaf culture values.
This past year in ASL II, Emelie and the other second year students continued to develop their receptive and expressive conversational signing skills while learning basic interpreting skills. The course was designed to increase conversational vocabulary, teach how to translate English sentences to ASL, incorporate more non-manual behaviors, classifiers, and gesturing in the students' signing, continue an awareness of Deaf culture values, and develop more 'voice to sign' interpreting skills. For the final project of the year students were required to translate a song into ASL.
In both ASL I and ASL II the students met once a week for 2 hours of class. They learned through games, role playing, songs, skits, field trips (some mandatory, some voluntary), and DVDs. Then, students were expected to complete their homework, including however much study time they needed to learn the material for that particular week.
In previous posts I have mentioned that Emelie is in the process of learning ASL. It seems to have sparked some interest because I have received a number of emails asking whether or not she likes taking ASL, what initiated her interest, if her coursework is done online, whether or not she plans to use it for her future, etc…so today I decided to let her speak for herself.
Please note that Emelie has a final year of American Sign Language (ASL III) to complete and recognizes she has years of learning ahead of her. Still, we are proud of her–she has worked hard to get to this point and is excited at the opportunity to answer questions that may spark a desire in another future student to get involved with, and learn, ASL.
Me: When do you first remember being interested in ASL?
Emelie: Ummm…I think it was 4 years ago in Missouri with Gram and Poppy. I was watching Gram sign some of the music they played during a worship service and thought that was kind of cool.
Me: Do you remember how you felt the first day of your ASL I class?
Emelie: I was excited and since I already knew the alphabet and numbers in sign language, I was feeling pretty confident.
Me: Did you lose confidence at any point after that first class?
Emelie: The last part of my first year I started losing a little bit of confidence after I took a test and made a C. There were a lot of signs in that one unit.
Me: What made those particular signs so difficult to learn?
Emelie: It was more of a grammar-type test I guess. There was just a whole bunch of signs. My problem wasn't remembering the signs exactly, but grasping the concept of the grammar. It is so much different than English and it was difficult because we didn't converse much at that point in the year–we were just trying to get through that one unit so I didn't have time to learn it with my hands.
Me: What has been your favorite part of learning ASL?
Emelie: Probably the Deaf events. I still get a little bit nervous beforehand because I don't know what to expect, but that is also what makes it my favorite part. Each time I attend a Deaf event I get to meet new people. Usually there are only a few people there, but twice there have been 20 or more people and I get so excited I want to pee on myself *giggle* — because it is so fun.
Me: You do realize I am going to include that you get so excited you want to pee on yourself, right?
Emelie: *eye roll* Believe me, I know.
Me: What exactly constitutes a Deaf event?
Emelie: A required part of our course. We have to go and participate in social activities with the Deaf – whether it is just hanging out at a Starbucks chat or helping with make-up and costume backstage for a Deaf Christmas play. A Deaf event is basically anything that a bunch of Deaf may be getting together for.
Me: What is the purpose of you attending Deaf events?
Emelie: We study a lot of Deaf culture, but you can't really get a grasp of it without being involved in it or being around Deaf and understanding the language and just how they treat each other. It helps students learn more about the language and the culture. Also, we have to write a paper on the event and get graded – ha! It's fun though – like writing a blog post.
Me: What is your biggest obstacle so far in learning ASL?
Emelie: The fact that I don't know anyone who is Deaf and since we live in a small town there aren't local Deaf events. I think when I finish ASL III it will be difficult for me to continue being in the Deaf culture without that.
Me: Right, which is why we've talked with Mrs. Karen about enrolling you in an Interpreting Training program through a local college after ASL III.
Emelie: Yes, I'm excited about that. I like to interpret and the little bit that I have done this year I have really enjoyed. It is difficult, but in a really enjoyable way.
Me: What do you think of Mrs. Karen, your ASL teacher?
Emelie: I LOVE MRS. KAREN. She reminds me of a mixture of two people I know and love. She really is something. She is strict, but not in a dull, boring way. She has a passion for ASL and the Deaf culture so it really encourages us to do better and learn more. She's…Mrs. Karen. I can't imagine having any other teacher but her.
Me: Tell me about the difficulty in interpreting the song for your final project this year.
Emelie: It was more time consuming than anything else. Just because of the grammar change, I think. But it was also my first time, so Mrs. Karen helped us more than she usually would. After I interpreted that song I interpreted another, which only took me two hours. A lot of the signs…well, when you interpret, each word doesn't have its own sign, so it is kind of like paraphrasing in a way. That takes up a little time–trying to find the right signs, but it is easier than interpreting what a hearing person is saying in sign language.
Me: What would you advise parents of kids interested in learning ASL?
Emelie: For one, do not, and I mean DO NOT try to learn solely off of the internet because there is so much junk out there that isn't really ASL. Recently I met a woman who is learning sign language and she taught herself from a bunch of websites she found online. Not everything you find on the internet about ASL is right and she ended up signing many things incorrectly–her grammar was off. There are things you can't learn from the internet like mouth morphing and classifiers.
Even though she had passion for ASL, she was learning incorrectly so that may prevent her accomplishing some things she wants to do in the future.
Me: But if that is the only way people can learn – as in, they can't drive an hour and 15 minutes like we do, what would you suggest?
Emelie: Yes, we are very blessed to be able to drive that far for my class, but for those who cannot…I would try to find someone who knows their stuff and see if you can get some resources from them.
Me: What do you see yourself doing in the future with ASL?
Emelie: I want to be an ASL interpreter.
Me: What exactly does that "look" like?
Emelie: I probably want to work for an interpreting agency. That is what Mrs. Karen has been doing for many years after her mission work with the Deaf. They send her to different places for different interpreting jobs and she said it is a very flexible job and schedule. Which I think I would appreciate with some of the other things I want to do with my future.
Me: Is there anything else you want to share about your experience learning ASL?
Emelie: It helps to make connections with people who also share an interest in ASL. I have three friends who are learning ASL with me and they've gone through 2 years with me and will finish out the 3rd year with me as well.
It's an encouragement just to have people with the same passion as you. Also, you have to love your teacher. It is a big part of it. I really look up to Mrs. Karen and sometimes my friends pick on me and say it looks like I'm absorbing everything she says but I just want to learn everything she knows.
Me: How do you communicate with those friends using ASL since you are so far apart physically/geographically?
Emelie: Usually we Skype or FaceTime.
The ASL I and ASL II classes have been a significant commitment for Emelie in the way of preparation/studying. She takes classes at a Homeschool Co-op that is more than an hour away from our home. This means ASL is also a significant commitment for our family.
Still, she has pursued ASL with fervor and we want to give her every opportunity to seek out her desired future and work toward that end. It also makes us hyper aware of what Meredith and Kenny are finding interest in now–wondering if their current passions will expand in the same way Emelie's has.
If you would like to see the song Emelie interpreted for her final project, "'The Best Thing About Me Is You' Ricky Martin ft. Joss Stone " you can view it above or at Vimeo. Inspirational Credit: lankylistman
Want More? A reader here at The Pioneer Woman suggested KellyMGreer, kmklined, as a resource for additional quality YouTube ASL videos.
What passions have your children pursued/are currently pursuing? If a graduate, how has that passion played out in their lives?
About the Author: Tired of feeling overwhelmed, Heather Sanders and her husband Jeff faithsized their family into a 960 square foot lake cabin in need of renovation. She writes about faith and simplifying your desires so you can be content right here and right now.
120 Comments and 15 Replies
Mandy D. On Saturday, April 28 at 8:03 am
She is so awesome!!! Love her own unique style!!! Keep it up Emelie, you will inspire many!!!
Chelsea On Saturday, April 28 at 8:59 am
I heart Emilie. She is one cool teenager.
Briana On Saturday, April 28 at 12:56 pm
That was beautiful. I am also learning ASL, on my second semester, and will definitely share this video with my 9yr old daughter who is learning along with me. Thank you for sharing.
joanne On Saturday, April 28 at 7:06 pm
oh heather, i just love emilie, she is a true original!! i always enjoy her videos and this one does not disappoint! she's awesome and you have done a great job with your children….joanne (retired homeschool mom)
Sheila On Sunday, April 29 at 12:39 am
This is so exciting to see someone young excited about Deaf culture and American Sign Language. I love that she is not only learning ASL, but is being paired with something just as, if not more, important: the culture. I went to California State University, Northridge because they have an excellent Deaf Studies program and had the joy and pleasure of having Dr. Lawrence Fleischer as a professor before he passed away, along with many other phenomenal instructors. The program, to my knowledge, is still 2nd best, with only Gallaudet coming first for Deaf Studies programs. It may be worth checking out if this is a path she wants to really pursue. There's a large Deaf culture here in LA (and San Diego) and it's an excellent program and experience.
Saffron G On Sunday, April 29 at 3:38 pm
I went to Makaton classes. Makaton is a form of sign language for disabled children, it was recommended as my son is disabled and wasn't supposed to speak. 3 days after I had finished my extensive course, he spoke which I sort expected to happen to be honest. I have know found 10 years later I barely remember it. So I hope Emelie should use her skill regualarly, use it or lose it!!
Sarah On Sunday, April 29 at 9:18 pm
Emelie, what an amazing and inspirational young woman you are!!!
Pejibaye On Monday, April 30 at 2:59 am
Emelie — I read this post a few days ago, while I've been traveling in Japan. This morning, I saw two women signing at the railroad station and thought: "I wonder how it's related to ASL? Could Emelie understand them?"
Sid On Monday, April 30 at 4:14 am
Emelie, you ROCK!
While there are excellent programs out there in many cities, Gallaudent University is TOPS if you think you might seriously want to pursue a career with ASL.
What better place can you go where there are thousands of Deaf people?
Keep up the great work!
Gina On Monday, April 30 at 2:23 pm
I also have had the desire to learn ASL, I just think it is so great. We are having the opportunity with my son right now as he has had some speech delays, it has helped a lot. Great job, I hope you can keep it up.
JudyLynn On Monday, April 30 at 3:19 pm
Excellent job, Emile!
To answer your question – how early passions play out – our youngest daughter (we live in California) went on a week long trip to Washington DC in 8th grade. It had a profound effect on her. After that trip she became single minded about government, history and politics (my husband and I have science degrees). As a Junior in HS she was honored to be chosen to work as a Congressional Page in the House while continuing her studies in classrooms on the top floor of the Library of Congress. It was an awesome experience and she discovered she also had an interest in foreign affairs. During college (where her major became Political Science) she spent a semester abroad in Russia. She graduated college in May 2011 and times being as they are had problems finding work. We have always encouraged her to grow and follow her passions. She is now working in DC – a few blocks from the Capitol – for a non-profit NGO that specializes in programs assisting the poor in other countries. I am totally amazed!
beautiful 2x On Tuesday, May 1 at 10:47 am
In one of my previous comments I mentioned that I am rarely happy with ASL interpretations of English songs (this may be really more of a personal quirk than any truly valid reason – I am very fluent in the English language, it being my first language, actually, so I often feel more connected to the English version than the ASL version). Well, today I came across a Youtube channel of a hearing person signing songs using ASL who changed my mind. His ASL is not very fluent YET he is so entertaining and I loved his facial expressions and great attitude of fun; he approaches the whole thing very creatively and intelligently. Please check out CaptainValor on Youtube Seems he gained fame (or infamy? with 'Party in the USA." It's all about intent – as long as one is coming from a place of openness and joy one certainly can do no wrong! Exactly like you, Emelie. I hope you enjoy CaptainValor as much as I do. To me his ASL is pretty good!
beautiful 2x On Tuesday, May 1 at 11:01 am
Incidentally, the largest deaf-blind community is Seattle, WA (they flock there because apparently the light quality of the Pacific Northwest is easiest on their vision – those who still have a bit of vision). So that is one avenue of interpreting (deaf-blind signers communicate tactically by very lightly and barely touching the signing person's hands). I agree that Gallaudet is the most popular option, however, CSUN (Northridge, CA) and RIT (Rochester, near Buffalo, NY, where NTID – National Technical Institute for the Deaf – is located) are equally as good and more open-minded/flexible/tolerant in some ways. TX options are, eh. Lamar University in Beaumont, TX, is the closest decent option. Austin Community College is great – Austin is fast becoming a very large deaf population center; lots of Gallaudet graduates are moving there so there is a strong culturally Deaf community there.
Karenliz On Tuesday, May 1 at 3:13 pm
Very impressive. My daughter graduated as a license ASL Interpreter from Columbia College Chicago in May 2011. She has worked as an interpreter right away. She fell in love with ASL by going to see a play and there were ASL interpreters there. She said they were better than the actors. She loves it. She now has a few clients that are death blind and although it was tough in the beginning her clients love you and request her now. She is now awaiting her scores for a licensing exam which will move her license up a notch. Her goal is to interpret theater. She said its very demanding, frustrating but so rewarding. Keep up it up!
cc On Wednesday, May 2 at 1:15 am
I'm hoping ASL will fit into my curriculum at school as an elective and that it'll be transferrable. If not I plan on taking it anyway. Possibly the Spring 2013 semester.
My cousin is deaf and I learned a lot when I was younger. I still remember the alphabet and a few other signs, though sometimes they completely slip my mind.
I'm looking forward to the challenge of learning ASL. I've been told that it's not an easy class.
Also, I think it's great that Emelie is learning it. She's an awesome kid.
Jenn Oh On Thursday, May 3 at 12:09 am
Emelie: I like your video! I took ASL years ago and loved it too! You are good!
Baylee Barnes On Thursday, May 3 at 6:25 am
I want to be an ASL interpreter as well but I am scared because I am just now trying to find classes this summer. If anyone has any answers for me, like which is the best ASL program to take in Texas.I would love that. Emelie, you did a wonderful job and I am so bless that my mom found this and told me to look at it.
Thanks and have a bless day!
Lena On Friday, May 4 at 3:55 pm
Wow. This is SO great. I've definitely been toying with the idea of taking some real sign language classes (since I started teaching my children how to sign–been using SignShine if anyone is interested!). I'm going to have to start looking into it some more. You've inspired me! ASL to me is one the most beautiful languages–it can do good things for everyone and express so much emotion. Beautiful. Keep up the good work.
Kelli On Friday, May 4 at 8:10 pm
My daughter is also learning ASL, here in Fl we can count it as her foreign language. A local church offers classes (as they have deaf members and interpreters in the congregation) so several homeschool families attend together It's hard work and Emelie is doing an awesome job!
Katy Belle On Sunday, May 27 at 8:36 am
I'm coming here to comment weeks after you posted because Emelie has stuck in my mind! I found this post, and Emelie's video very interesting! I had never thought about the importance of body language, facial expression, etc.
At my sons' Middle School they employ 2 ladies full time to sign for 3 students. At the recent Awards Ceremonies and 8th grade Farewell Assembly, I sat near these ladies and watched. Amazing! I loved the facial expressions and extra stuff they put in there. What I REALLY loved is that it was obvious that they were trying to convey the WHOLE experience for these girls! When kids would scream in the audience as a certain name was ready, she would sign the scream! When a random girl would scream "Madison I love you!" as her friends walked across the stage, she signed that! She attempted to sign the WHOLE experience. Of course. But I never thought about it! I'm not sure, but I think that when the choir was singing, she even indicated when it was just the male voices and just the female voices! Very impressive and such a blessing for the people that need her! They deserve the WHOLE Middle School experience! My son, 14, loves to tell jokes and the interpreter always signs them!
AND… We recently went to a Van Halen concert and I noticed a lady signing to one or more people. It cracked me up to realize how few words are in most songs. Watching her sign the song "Jump" was so fun! She was jumping a lot! Again, the body language!
Anyway…Emelie, you opened my eyes to something that has been in front of me that I paid little attention to. Thank you!


text:
My brother just texted me that he was hanging with Nolan. NBD.
My brother just texted me that he was hanging with Nolan. NBD.
Ask Nolan if he voted for himself in the poll.
Last edited by CBT; 05-01-2012 at 03:29 PM.
I feel bad for Feliz. He's going to have to be perfect today with the lineup we're trotting out.
Will Clark
Julio Franco
Dean Palmer
Alex Rodriguez
Ivan Rodriguez
OUTFIELDERS (choose 3)
Juan Gonzalez
Rusty Greer
Oddibe McDowell
STARTING PITCHERS (choose 2)
Kenny Rogers
Nolan Ryan
RELIEF PITCHERS (choose 2)
Neftali Feliz
John Wetteland
Rafael Palmeiro
Will Clark
Mike Hargrove
Pete O'Brien
Rafael Palmeiro
Mark Teixeira
Julio Franco
Ian Kinsler
Mark McLemore
Bump Wills
Michael Young
Buddy Bell
Hank Blalock
Steve Buechele
Toby Harrah
Dean Palmer
Elvis Andrus
Scott Fletcher
Toby Harrah
Alex Rodriguez
Michael Young
Rod Barajas
Gerald Laird
Geno Petralli
Ivan Rodriguez
Jim Sundberg
OUTFIELDERS (choose 3)
Juan Beniquez
Jeff Burroughs
Nelson Cruz
Juan Gonzalez
Rusty Greer
Josh Hamilton
Pete Incaviglia
Oddibe McDowell
Kevin Mench
David Murphy
Al Oliver
Larry Parrish
Bill Sample
Ruben Sierra
Gary Ward
George Wright
STARTING PITCHERS (choose 2)
Kevin Brown
Jose Guzman
Rick Helling
Charlie Hough
Ferguson Jenkins
Jon Matlack
George Medich
Kevin Millwood
Darren Oliver
Roger Pavlik
Gaylord Perry
Kenny Rogers
Nolan Ryan
Bobby Witt
RELIEF PITCHERS (choose 2)
Francisco Cordero
Danny Darwin
Neftali Feliz
Frank Francisco
Tom Henke
Kenny Rogers
Jeff Russell
John Wetteland
Mitch Williams
C.J. Wilson
Brian Downing
Juan Gonzalez
Tom Grieve
Rafael Palmeiro
Larry Parrish
Got Dammit the offseason sucks.....
oh, $#@! wait....
I know Texas doesn't have Josh or Beltre playing tonight, so it looks like a pretty lack luster lineup for us(spoiled Rangers fans? Yep)
But how much you want to bet that 90% of the league would trade their roster for the lineup that Texas is throwing out there tonight?
I bet the San Fransisco Giants and Phillies would KILL to have these guys behind their Starting Pitchers...and that's to say nother for the A's and Mariners in their own division.
Oh, I bet the Angels would love to swap lineups right now too, LFMAO.
glad that half inning is over.
Not a good start...
Watch this young, no name Pitcher destroy our lineup...
And Nefti allows a lead off hitter, ugh.
BIE(Best Infeild EVAH) with the quick and easy DP.
Nice K by Nefti...almost made that last at bat look easy.
Last edited by drew8117; 05-01-2012 at 06:19 PM.
Ummm holy $#@! either tali is feeling it or the gun is hot. He's hittin upper 90s consistently tonight
Not starting off well for our hitters...
And then Murph gets a nice opposite field hit.
Time for Nap to do something.
C'mon Mitchy...a hit scores at least one.
But watch, after that Homer last night, he'll obviously hit a DP ball.
mitch is getting after it.
Dinking and dunking them tonight...this is the way they'll score, lol.
How was that not a DP?!?
I love this Rangers team.
No Hamilton...No Beltre. Andrus, Kinsler, Face, and Cruz don't do $#@!.
It's okay...Gonzales and Gentry will get you RBI's.
i'm not disagreeing with you.
i'm just pointing out that you have a very jovial approach to your internet speak.
Easy steal...didn't even attempt a throw.
Wow...what a start for the back of the Rangers Lineup.
When I saw the lineup, it looked pretty weak back there...but they showed up pretty well so far.
happy trails soon.
Is it okay to feel bad for the opposing pitcher?
"Hey, new kid...here's what I want you to do early in your career...go out and shut down the Best Team in Baseball. Oh don't worry, they aren't playing MVP Josh Hamilton, or another big bopper in Beltre...you'll be fine if you get past Cruz and Napoli!"
And these aren't really strong hits...a bunch of bloopers and soft hit balls right now.
Leave it to Face.
BTW, how many runs were scored with 2 outs? I know Texas has a nack of scoring in bunches when they have 2 outs.
Oh $#@!!
The just said if everything goes right, Yu will start on Sunday...then start next Friday against CJ F'ing Wilson?!?
That's must see right there...CJ will be going for the shut out, you know that, and Yu won't give a $#@!, and still probably out pitch him.
Not a good start to the 3rd...with two walks, and now a $#@!ing HR....
C'mon Nefti...you're getting sloppy.
Craig looked like he got hurt too.
And what went from a great day for Nefit, and the Rangers, looked like it was going to be easy honestly...now looks like they'll have to work hard for it.
Last edited by drew8117; 05-01-2012 at 07:09 PM.
$#@!ing $#@! happy.
$#@!ing $#@! craig.
Goddamnit feliz. Quit $#@!ing walking people, especially with a five run lead. Christ
It'll be nice to see Colby Lewis, and his consistent self tommorrow.
Feliz looks rattled now.
let's go ahead and lower that pitch location please, happy.
damn when JB gets all of it he doesn't f around. might be one of the biggest 'no doubters' in the league. f'n batting practice pitch from NF.
there you alberto. let's $#@!ing bat a little.
Wow, if Toronto can hang onto the ball, he's out...twice!
One of those "Oh no, Oh no, Oh no...YES!!" plays.
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