text:
|Publication number
|Publication type
|Publication date
|Apr 1, 1975
|Filing date
|Jan 30, 1974
|Priority date
|Jan 30, 1974
|Also published as
|Publication number
|Gordon L Dorn, Joseph M Hill
|Original Assignee
|Wadley Res Inst & Blood Bank
|Export Citation
|BiBTeX, EndNote, RefMan
|Patent Citations (5), Referenced by (50), Classifications (20)
|External Links: USPTO, USPTO Assignment, Espacenet
United States Patent [191 Dorn et a1.
[ 1 Apr. 1,1975
[ APPARATUS AND METHOD FOR THE DETECTION OF MICROBIAL PATHOGENS [75] Inventors: Gordon L. Dorn; Joseph M. Hill,
both of Dallas, Tex.
[73] Assignee: J. K. and Susie L. Wadley Research Institute and Blood Bank, Dallas, Tex.
[22] Filed: Jan. 30, 1974 [21] Appl. No.: 437,890
[52] US. Cl....,. l95/l03.5 R, 128/218 M, 128/272, 206/222, 215/247, 215/D1G. 3, 210/D1G. 23, 233/26, 195/127 Pllllltll) E.\um1'nerAlvin E. Tanenholtz Ans-[slum bli'aminerRobert J. Warden Almrney, Agent. or FirmRichards, Harris & Medlock [57] ABSTRACT An apparatus for the isolation and concentration of microbial pathogens from a sample fluid which comprises an elongated enclosed centrifugation receptacle containing an evacuated space in contact with a sterile aqueous solution of a material having a greater density than the sample fluid but able to selectively receive microbial pathogens from the sample fluid, a first injectable closure on one end thereof and a second injectable closure on its other end, and a sterile end chamber in communication with the second injectable closure which contains a sample fluid treating solution together with an injection needle assembly which when actuated will pass through the second injectable closure means and allow communication between the chamber and the evacuated space within the centrifugation receptacle to thereby allow the treating fluid to flow from the chamber into the evacuated space. Thereafter, the sample fluid can be injected into the evacuated chamber of the centrifugation tube through a needle passing through the first injectable closure and allow the sample fluid to admix with the pretreating solution in the evacuated chamber. The article is then subjected to centrifugation to allow the microbial pathogens to selectively pass into the liquid filter medium.
25 Claims, 6 Drawing Figures Pf-IENTEDAPR 1 I97 FIG. 2
i ii/ FIG-4 APPARATUS AND METHOD FOR THE DETECTION OF MICROBIAL PATHOGENS This invention relates to the detection of microbial pathogens. In another aspect, this invention relates to a novel apparatus which is used for admixing a sample fluid containing microbial pathogens with a treating fluid and thereafter selectively extracting the microorganisms from the sample fluid. In still another aspect, this invention relates to a novel method and apparatus for diagnosing septicemia.
Scpticemia, which is the presence of pathogenic microorganisms in the blood, is one of the most serious types of infection encountered. in spite of an armament of antibiotics and fungal drugs, the mortality rate from septicemia is approximately 2571. ln addition, when shock accompanies septicemia, the mortality rate increases to over 60%. Patients who are suffering from debilitating diseases, undergoing major surgery, receiving immunosuppressive drugs. or anticancer medications are particularly prone to septicemia.
Early administration of appropriate antibiotic therapy is important in fighting septicemia. Consequently, it is imperative that the physician known as rapidly possible not only whether the patient has septicemia, but also the identity of the infecting microorganisms, and the susceptibility of the microorganisms to antibiotic agents. Thus, the proper diagnosis of septicemia depends upon very rapid and efficient quantititative analysis of the patients blood. It is imperative during the quantitative analysis of the patient's blood or other body lluid that the sample fluid not be contaminated with pathogens from the laboratory environment.
Three analytical systems have been conventionally utilized to determine the presence of microorganisms in a body fluid. These conventional systems include the liquid broth culture technique. the so-called pour plate method, and the filtration method using a solid matrix filter. Each of these systems has its drawbacks, and none of the system provides a rapid detection of microorganisms in the blood sample. Generally, the liquid broth method is not quantitative, the pour plate method and the filtration method using a solid matrix filter are open systems subject to external contamination, e.g., the introduction of pathogens onto the culture by the laboratory atmosphere and personnel.
Recently, an improved method and apparatus have been developed for determining the presence of microbial organisms within a sample fluid. This improved method is very rapid and quantitative. Such method is disclosed in copending U.S. Pat. application Ser. No. 428135. filed Jan. 9, 1974 (Attorneys docket No. B- 2532C which is a continuation-in-part of US. Pat. application Ser. No. 331,693. filed Feb. l2, l973, and entitled Deteetion ofthe Microbial Pathogens. According to this improved method for detection of microbial pathogens, a sample of body fluid such as blood (preferably a lysed blood sample) is deposited upon a liquid filter medium within a confined, sterile Zone. The liquid filter medium has a density greater than the sample fluid and comprises a sterile aqueous solution which will selectively receive microbial pathogens from the sample fluid. Thereafter, the confined sterile zone is subjected to centrifugation to force the sample fluid against the liquid filter medium and cause microbial pathogens to selectively pass therein and thereby separate from the mass of the body fluid sample. Next, the
liquid filter medium containing the microbial pathogens is separated from the remainder of the sample fluid and portions of the liquid filter medium are subjected to culturing conditions.
The improved method described above does provide a very rapid and efficient procedure for separating microbial pathogens from a sample fluid. However, in the preferred method, the sample fluid is admixed with lysing and anticoagulant agents prior to the time that it is injected into the evacuated centrifugation zone containing the liquid filter medium. Some liquid filter mediums are incompatible with some of the pretreating and/or lysing agents. lt has not heretofore been possible to combine the two materials for long periods in the sterilized centrifugation tube. Therefore, it has been necessary to subject the sample fluid to the possibility of external contamination by the additional mixing step which of course means an additional entry into the closed system.
According to one embodiment of the subject invention, an apparatus is provided for use in the improved method described above which will aseptically introduce the lysing agent into the evacuated centrifugation tube containing the liquid filter medium and thereby allow the blood sample or sample fluid to admix therewith just prior to the centrifugation step wherein the microbial pathogens are selectively removed from the remainder of the sample fluid by the liquid filter medium. The improved apparatus of the subject invention generally comprises an elongated centrifugation tube containing an evacuated centrifugation chamber having a first end and a second end; a first injectable closure means sealably closing the first end thereof and a second injectable closure means sealably closing the second end thereof; and an enclosed end chamber containing a treating fluid such as a lysing agent is affixed on the second end of said elongated centrifugation vessel in communication with said second injectable closure means, said end chamber containing an injection means to aseptically transfer said treatment fluid from said end chamber chamber through said second injectablc closure means to said centrifugation chamber. In a specific embodiment said injection means comprises a tubular stylus positioned for reciprocal movement in said end chamber and has a first opening adjacent its leading end and a second opening in its barrel such that when advanced toward said second injectable closure means in said chamber, the stylus will pass through the second injcctable closure means and its first opening will communicate with the evacuated space while its second opening maintains communication with said end chamber; and means to actuate said stylus means.
In a preferred embodiment of the subject invention, the chamber containing the evacuated space also contains an aqueous filter medium having a greater density than the sample fluid and which will selectively receive the microbial pathogens from the sample fluid, and the end chamber contains an aqueous solution of a lysing agent such as purified saponin extract and optionally other materials such as an anticoagulant and/or an oxygen scavenger such as ascorbic acid and/or a thioglycolate. It is preferred that such materials be sufficiently stable so that they will not decompose or be otherwise deleteriously affected under autoclaving conditions.
In accordance with another preferred embodiment of this invention, the above-described centrifugation and mixing apparatus is provided with a syringe means for removing the aqueous filter solution from the evacuated chamber which includes a stylus sufficiently long to only penetrate the thickness of the first injectible closure means and not to materially pass into said aqueous filter medium.
This invention can be more easily understood from a study of the drawings in which:
FIG. 1 is a sectional view of a preferred centrifugation and mixing apparatus of the subject invention;
FIG. 2 is a sectional view of the apparatus of FIG. 1 showing the stylus in the upper chamber in actuated position;
FIG. 3 is a sectional view of the apparatus in FIG. 1 showing a sample fluid being injected therein through the bottom closure:
FIG. 4 is a sectional view of the apparatus of FIG. 3 showing the aqueous filter medium being removed via a needle passing through the lower closure;
FIG. 5 is a sectional view of another embodiment of the subject invention; and
FIG. 6 is a sectional view of the embodiment of FIG.
5 showing the stylus in actuated position.
The improved centrifugation and mixing apparatus of the subject invention provides an improved means for carrying out the method of detecting microbial pathogens which is set forth in copending patent application Ser. No. 428,135 filed Jan. 9, I974, (Attorneys docket No. B2532C) which is a continuation-in-part of US Pat. application Ser. No. 33l ,693, filed Feb. 12, I973, and such disclosure is herein incorporated by reference into the subject application.
Now specifically referring to the drawings, the novel centrifugation and mixing apparatus as illustrated in FIGS. 1-4, will be described in detail. As shown in FIG. 1. the mixing and centrifugation apparatus 10 comprises an elongated, tubular centrifugation vessel 12, having an injectable closure member 14 which sealably closes the lower end thereof, and an injectable closure member 16 which sealably closes the upper end thereof. Furthermore, the upper end of injeetable closure member 16 is enclosed by cap 18. As shown, cap 18 carries screw threads 20 which operatively engage screw threads 22 positioned around the upper periphery of centrifugation vessel 12.
Cap 18 generally comprises a cylindrical body carrying screw threads 20 on the inside periphery of the opening thereof adjacent shoulder 24 which receives the outer rim ofinjectable closure member 16. Furthermore, the outer edge of shoulder 24 forms the opening of cavity 26, which is preferably a short cylindrical shape. The upper end of cavity 26 is enclosed by end wall 28. Rod 30 of push button member 32 extends through aperture 33 in end wall 28. Furthermore, rod 30 carries tubular stylus 34 on its leading end. Tubular stylus 34 comprises a sharp, beveled end 36 and an aperture 38 through its side wall as shown in the drawing. Furthermore, a resilient washer is positioned snugly around the upper end of tubular stylus 34 in a manner to seal the interior of cavity 26 above aperture 38 of tubular stylus 34.
Centrifugal vessel 12 can be made of glass or hard plastic such as polycarbonate or polypropylene and cap 18 can be made of a hard plastic. Injectible closure member 14 can comprise a rubber'self-scaling stopper. The leading end of injectable closure member 14 carries a frustoconical recess 14a. Injectable web 14h forms the bottom of the frusto-conical recess 14a. In;
jectable closure member 16 can also comprise an injectable rubber self-sealing stopper. The top of injectable closure member 16 carries a recess 16a, and the leading end of injectable closure member 16 carries a recess 16b. Recesses 16a and 16b are separated by injectable web 16c. If desired, injectable closure members 14 and 16 can be of the same basic configuration. It is only desirable that in jectable closure member 16 carry recess 16a on its top portion and that injcctable closure member 14 carry a frusto conical recess 14a within the leading end thereof. The sterile contents of centrifugation vessel 12 comprises liquid filter medium 42 and an evacuated space 44 which may be a complete or a partial vacuum. Space 44 is maintained at a lower than atmospheric pressure at a predetermined value so that the centrifugation vessel can receive a known amount of liquid by injection through injectable closure members 14 and 16 without excessive pressure being built up within the interior thereof which could cause injectable closure members 14 and 16 to become dislodged from the openings within centrifugation vessel [2.
The liquid filter medium 42 can be of any of the liquid filter media set forth in said copending patent application for detecting microbial pathogens and generally, comprises an aqueous solution of any solute which is nontoxic to the microbial organisms being suspended,'
and has a density sufficient high to suspend red and white blood cells or blood cell debris. The solute is preferably nonionic. Thus, the liquid filter medium has a density greater than blood, e.g., greater than about 1.06 gm/cc, and will suspend blood cells or blood cell debris, but yet will receive microbial pathogens. In addition, the liquid filter medium preferably contains a minor amount of a thermally sensitive gelling agent.
Suitable solutes which can be used in the liquid filter medium 42 include the sugars such as sucrose, glucose, maltose, fructose, manitol, sorbitol, and the like. Generally, liquid filter medium 42 should be at least about 40 wt 7( of the sugar and can contain the sugar up until the saturation limit thereof. Preferably, the sugars are contained within liquid filter medium 42 in the range of from about 50 wt 71 thereof. Generally, the sugars, and especially sucrose, are preferred solutes for liquid filter medium 42 because the liquid filter medium can be maintained at a physiological pH, i.e., 6.0-7.0 and when combined with gelatin, they can be autoclaved. Any solute can be used in the scope of this invention so long as the resulting solution is more dense than red blood cells and red blood cell debris, and is nontoxic to the microbial pathogens. Other suitable such materials include a chemical commonly known as Hypaque sodium, C H I N NaO (3,5 -diacetamido-2,4,6- triidobenzoic acid sodium salt). This material can be utilized in aqueous solution in the same concentration as the sugar as described above. Another class of solutes which can be used to form the aqueous liquid filter medium in the scope of the subject invention includes macromolecular solutes which are capable of producing a liquid gel structure in aqueous media which have a pour size small enough to preclude red cells or red cell debris but large enough to pass microbial pathogens.
An example ofa suitable such macromolecular solute is a water soluble crosslinked polymer having microporous openings throughout its solubilized network. A suitable such water soluble polymer includes a copolymer of sucrose and epichlorohydrin which has a weight average molecular weight in the range of from about 300.000 to about 500.000, and an intrinsic viscosity of about 0.1 7dl/g, a specific rotation lal of +56.5 and contains dialyzable material in an amount of less than 1 weight percent. A suitable such polymer is sold under the trademark of FICOLL by Pharmacia Fine Chemicals, Inc. 800 Centennial Avenue, Piscastaway, NJ. Another such polymer which can be used in the scope of this invention is dextran, having a weight average molecular weight in the range of about 10,000 to about 2,000,000 and preferably about 50,000 These polymers, when dissolved in water in accordance with the subject invention function as a liquid filter medium for microbial pathogens and apparently have microporous openings throughout the solubilized network in the range of from about 1 micron to about 7 microns.
The water soluble polymer or macromolecular solute is preferably present in the aqueous solution in the range of from about 10 to about 40 wt "/2 and more preferably from about to about wt 7r thereof.
It is to be understood that the term thermally sensitive gelling agent is meant any agent which will gel the aqueous solution of filter medium 42 at a temperature generally lower than room temperature but yet will liquefy at higher temperatures which are nondeleterious to the microbial pathogens. c.g., lower than about 50C and generally no higher than about 42C. Suitable thermosensitive gelling agents include any such gelling agent which is nondeleterious to the solution or to the sample being analyzed. Examples of suitable such materials include gelatins, i.e., the proteins obtained from collagen by boiling skin, ligaments, tendons, bones, and the like in water. Any suitable amount of thermally sensitive gelling agent can be utilized, e.g., about 0.5 to about 5 wt Z of filter medium 42.
Furthermore. it should be noted that cavity 26 can be any convenient shape and preferably has a round cross sectional area and is a short cylindrical shape and is generally in alignment with recess 16a ofinjectable closure member 16. Thus, tubular stylus 34 which is carried by rod 30 of push button member 32 will axially move within cavity 26, and resilient washer will maintain the upper portion of cavity 26 sealed at a point above aperture 38 of tubular stylus 34. Furthermore. it is noted that the spacing between aperture 38 and beveled end 36 of tubular stylus 34 is slightly greater than the thickness of injectable web 16c such that when push button 32 is fully extended, the opening adjacent beveled and 36 will pass through injectable web I6c and be positioned within space 44 ofcentrifugation vessel 12 while aperture 38 will communicate with cavity 26 as illustrated in FIG. 2. This position of tubular stylus 34 will allow liquid 46 which is contained within or carried by recess 16a to pass into centrifugation vessel 12 by action of the lower pressure within space 44 and pass via aperture 38, the tubular body of stylus 34 and opening adjacent bevel point 16a.
Treating solution 46 can contain any suitable ingredient or ingredients with which it is desired to treat the sample fluid before microbial pathogens are separated therefrom. In accordance with a specific embodiment ofthe subject invention. treating solution 46 comprises an aqueous solution of a lysing agent for blood. Any suitable lysing agent can be utilized in the aqueous solution which is nontoxic to microorganisms. A suitable such lysing agent is a nontoxic aqueous solution of saponin. It must be noted that many saponins are thought to be toxic to microbial pathogens. However, as set forth in copending application Ser. No. 423,447. filed DEC. 10, I973 (Attorneys Docket B-2774) entitled DETOXIFICATION OF SAPONINS, which is herein incorporated by reference into this application, a new method is disclosed for removing the toxic ingredients from the heretofore thought to be toxic saponins. In general, the toxic saponin material can be detoxified in accordance with the invention set forth in this copending patent application and the resulting purified material used in the scope of this invention. Some commercially available saponin preparations are nontoxic, and of course, the materials can also be used in the scope of the subject invention. In addition, the aqueous solution can contain an anticoagulant and/or an oxygen scavenger. A preferred anticoagulant is sodium polyanethol sulfonate (SP8) or Heparin, for exmaple. Sodium polyanethol sulfonate is preferred because it not only acts as an anticoagulant but also inhibits the phagocytic activity of granulocytes and monocytes and the normal antibacterial activity of serum.
Now referring to FIGS. 1-4, the use of the apparatus as illustrated therein will be described in detail. Liquid filter medium 42 can comprise l.5 ml of an aqueous solution containing 50 wt '71 sucrose and 1.5 wt gelatin, for example.
Treating solution 46 can contain any such suitable constituent such as a lysing agent and/or an anticoagulant and/or an oxygen scavenger in any desirable concentration. Any amount of an anticoagulant which is sufficient for the amount of blood sample and any amount of lysing agent sufficient to lyse this blood sample, can be used, for example, 0.3 milliliters of an aqueous solution containing about 12% by weight of nontoxic saponin and about 2% by weight sodium polyanethol sulfonate. Initially, the centrifugation and mixing apparatus 10 is placed in an upright position as illustrated in FIG. 1 to allow liquid filter medium 42 to pass downwardly against injectable closure member 14. Next. apparatus 10 is placed in a suitable cooling unit such as a refrigerator and chilled sufficiently to cause the gelatin to solidify the liquid filter medium 42. For example. the tube can be chilled to 49C. Next, push button member 32 is depressed in a manner illustrated in FIG. 2 to thereby force beveled end 36 of tubular stylus 34 through injectable web such that the opening adjacent beveled end 36 communicates with evacuated space 44 and aperture 38 remains in communication with cavity 26. This action will subject the interior of cavity 26 to the lower pressure maintained within evacuated space 44 and this will therefore result in treating solution 46 passing through aperture 38, the barrel of tubular stylus 34, and from the opening adjacent the beveled end 36 into evacuated space 44 and become deposited as layer 46a upon the solidified liquid filter medium 42.
Next. a sample fluid such as a blood sample (e.g., 8 ml) is obtained within syringe 50 which carries hypodermic needle 52. The blood sample is then injected within the interior of centrifugation vessel 12 in a manner schematically illustrated in FIG. 3. Needle 52 pierces web 14b of injectable closure member 14, passes through the congealed aqueous liquid filter medium 42 and then the plunger of syringe 50. is depressed to deposit blood sample on the congealed liquid filter medium 42 and cause a turbulent mixing with treating solution 46 and form a resulting mixture 48. It is noted that solution 46 can be added to the blood sample after it has been injected into centrifugation vessel 12; however, it is preferred that it be added initially to promote better mixing with the blood sample. The turbulence caused by the blood passing into the evacuated space 44 and turbulently admixing with treating solution 46 will not disturb the liquid filter medium 42 which will remain as a solid bottom layer, as illustrated in FIG. 3. After this is accomplished, cap 18 can be unscrewed and removed from the upper portion of apparatus 10 as illustrated in FIG. 4.
The mixing of the blood sample with the treating solution 46 containing the lysing agent will result in the red blood cells becoming lysed which will therefore minimize the possible trapping effect or erythrocytes and/or lymphocytes. This trapping effect would in general comprise the erythrocytes or lymphocytes becoming stacked on top of the liquid filter medium during the centrifugation step and the stacked cells trapping microbial pathogens as they are passed downwardly during centrifugation and thereby .prevent them for reaching the liquid filter medium. Furthermore, the sodium polyanethol sulfonate within treating solution 46 acts as an anticoagulate and inhibits the phagocytic activity of granulocytes and monoeytes and the normal antibacterial activity of the serum once it becomes admixed with the blood sample.
Next, hypodermic needle 52 is withdrawn from the injcctablc closure member I4 and centrifugation vessel 12 containing the congealed liquid filter medium 42 and the blood sample admixed with treating solution 46 (illustrated as layer 48 in FIG. 3) is heated while in the upright position sufficiently to melt the gelatin and cause the liquid filter medium 42 to liquefy. Centrifugation vessel 12 is heated to a temperature which will not destroy any microbial pathogens which may be present in the blood sample but which will be sufficient to liquefy the gelatin. For example, while in the position as illustrated in the drawings, centrifugation vessel 12 can be heated by immersion in a water bath to a temperature set at about 37C-42C. Thus, the liquefication of the gelatin within the liquid filter medium 42 yields a liquefied aqueous filter solution which is now ready to function as a liquid filter medium for the microbial pathogens.
The separation of the microbial pathogens from the remaining portion of the blood sample is accomplished by placing centrifugation vessel I2 into a suitable centrifugation apparatus and subjecting it to sufficient centrifugal force to separate the microbial pathogens from the remaining constituents in the blood sample. The speed and time of centrifugation can vary widely depending upon the construction material of centrifugation vessel 12 and the type of centrifugation apparatus. The centrifugation can be conveniently accomplished by imparting between about 100 and about 6,000 gravities and preferably from about 1400 to 5000 gravities to centrifugation vessel 12. A suitable method would include a swinging bucket centrifuge rotor which imparts between 2000 and 4000 gravities for 10 to minutes to the particular system described in this preferred embodiment.
After centrifugation vessel 12 is subjected to the centrifugation step as described above, a sterile syringe 54 carrying a shortened hypodermic needle 56 is passed through injectable web 14b of injcctable closure member 14 as illustrated in FIG. 4. As illustrated. hypodermic needle 56 carries a stop 56a which will only allow the needle to pierce injectable web 14/; and the beveled end 36 to extend into liquid filter medium 42. After this is accomplished, syringe 54 withdraws liquid filter medium 42 from the interior of centrifugation vessel 12 to leave the residual sample solution mixture 48 therewithin.
It is noted that alternately a longer hypodermic needle can be utilized with syringe 54 and the needle can be passed to the interior of centrifugation vessel 12 to a point slightly past the interface between liquid filter medium 42 and the sample fluid layer 48 and the sample fluid layer initially removed from the tube.
In this instance. the syringe should initially have the plunger retracted and be filled with sterile air in order that the sterile air can be pumped into the interior of centrifugation vessel 12 incrementally as the sample layer is displaced. Thereafter, the liquid filter medium 42 can be removed.
The action of syringe 54 withdrawing liquid filter medium 42 results in the liquid filter medium and the microbial pathogens becoming thoroughly admixed and generally uniformly distributed therein. This liquid filter medium 42 containing the dispersed microbial pathogens is then distributed on suitable bacterial growth media. Suitable such growth media are set forth in said copending patent application entitled DETECTION OF MICROBIAL PATHOGENS.
For example. with l A milliliters of liquid filter medium containing microbial pathogens. one blood agar plate can receive 0.2 milliliters of the medium and the plate can be incubated at 37C in an aerobic atmosphere. Another blood agar plate can receive 0.2 milliliters of the aqueous solution and can be incubated at 37C in a candle jar. Another blood agar plate can receive 0.2 milliters of the aqueous solution and can be incubated at 37C in an anaerobic environment. Another 0.2 milliliters of the solution can be placed on a sabouraud agar plate and incubated at 25C in an aerobic environment. Another 0.2 milliliters of the solution can be placed on a EMB plate (Eosin methylene blue dye) plate and incubated at 37C in a candle jar. Another 0.5 milliliters of the solution can be placed in a liquid thioglycolate medium and incubated at 37C. The growth media can be checked daily for the presence of colonies. The number of microbial pathogens in l milliliter of blood can be determined by multiplying the number of colonies by a correction factor. This correction factor takes into consideration the recovery rate for a given organism, the volumes of blood and liquid filter solutions employed and the amount of final mixture plated. In the general example set forth above. the correction factor is 1.56.
Now referring to FIGS. 5 and 6. a centrifugation and mixing apparatus 10:: is illustrated which shows another embodiment of the subject invention. As shown, centrifugation apparatus 10,, is exactly the same as upparatus 10 except for cap 70. As shown in apparatus 10 cap comprises an enclosed cylindrical cap member having screw threads 72 on the inside walls thereof. and a tubular stylus 74 rigidly attached to the end wall 75 thereof. Tubular stylus 74 comprises the same basic configuration as tubular stylus 34 and comprises a beveled leading end 76 which carries an aperture and a second aperture 78 adjacent its midportion. As shown, with cap 70 in its first position as illustrated in FIG. 5, the beveled end 76 oftubular stylus 74 is positioned within treating solution 46 maintained within recess 16a of injectable closure member 16. in this position. it is preferred that a plastic strip 80 be positioned around the lower outside lip of cap 70. Plastic strip 80 can comprise a thermoplastic or thermosctting material which will deform such as by heat to conform to the lower outside lip of cap 70 and lower outside screw threads 22 carried by centrifugation vessel 12. This will assure that the closure member 70 will be maintained in its first position illustrated in FIG. 5 during storage and handling. lf it is desired to deposit the treating solution 46 upon liquid filter medium 42, cap 70 is then screwed downwardly to thereby deform strip 80 and force the beveled end 76 of tubular stylus 74 through the injectable closure member 16 as illustrated in FIG. 6. This will thereby allow communication between the cavity formed between recess 16a of injcctablc closure member 16 and end wall 75 and the evacuated space 44 and thereby cause treating fluid 46 to pass through aperture 78 through tubular stylus 74 and into the interior of centrifugation vessel 12.
While this invention has been described in relation to its preferred embodiments, it is to be understood that various modifications thereof will now be apparent to one skilled in the art from reading this specification and it is intended to cover such modifications as fall within the scope of the appended claims.
We claim:
I. An apparatus used for the isolation and concentration of microbial pathogens from a sample fluid comprising:
a. an enclosed centrifugation receptacle having a first end and a second end and containing an evacuated space maintained at a lower than atmospheric pressure adjacent a sterile aqueous solution of a material which is non-toxie to said microbial pathogens and has a greater density than the sample fluid but able to selectively receive microbial pathogens from said sample fluid and also containing a minor effective amount of a thermally sensitive gelling agent:
b. first injectable closure means sealably closing said first end of said elongated eentrifugation receptacle:
c. second injectahle closure means sealably closing the second end ofsaid elongated centrifugation receptacle;
d. a scaled end chamber in alignment with said second injectable closure means on said second end of said elongated centrifugation receptacle and containing a sample treating fluid sealed therewithin; and
e. injection needle means positioned within said sealed end chamber for passing said fluid from said end chamber through said second injectable closure means and into said evacuated space.
2. The apparatus of claim 1 wherein said injection needle means comprises a tubular stylus means positioned within said sealed end chamber in alignment with said second injcctable closure means and containing a first opening adjacent its leading end and a second opening through the sidewall thereof at a spaced distance therefrom. and means to advance said stylus means toward said second injectable closure means to a fully advanced position such that the leading end thereof and said first opening passes through said second injectable closure means and into said evacuated space and the second opening thereof remains in said second chamber.
3. The apparatus of claim 2 wherein said end chamber is formed between said second injectable closure means and a hollow cap means detachably affixed to said second end of said ccntrifugation vessel.
4. The apparatus of claim 3 wherein said cap means is threadably engaged with said second end of said centrifugation vessel.
5. The apparatus of claim 4 wherein said tubular stylus depends from said cap means in alignment in said end chamber with said second injectable closure means and is advanced to its said extended position by rotating said threadably engaged cap on said second end of said centrifugation vessel.
6. The apparatus of claim 4 wherein said tubular stylus is positioned on a plunger means in said end chamber in alignment with said second injectable closure means. I
7. The apparatus of claim 4 wherein said minor effective amount of said thermally sensitive gelling agent is from about 1 to about 5 wt /1 of said aqueous solution.
8. The apparatus of claim 7 wherein said thermally sensitive gelling agent is gelatin.
9. The apparatus of claim 8 wherein said aqueous solution is an aqueous solution of a sugar.
10. The apparatus of claim 9 wherein said sugar is sucrose.
11. The apparatus of claim 10 wherein said aqueous solution contains at least 40 wt /1 of said sucrose.
12. The apparatus of claim 8 wherein said aqueous solution is an aqueous solution of a macromolecular solute having microporous openings throughout its solubilized network, said openings being of sufficient size to selectively pass said pathogens from said sample fluid.
13. The apparatus of claim 12 wherein said polymer is epichlorohydrin-sucrose polymer having a molecular weight in a range of from about 300,000 to about 500,000 and a specific rotation [04],, of 56.5".
14. The apparatus of claim 12 wherein said polymer is dextran having a molecular weight of from about 10,000 to about 2.000.000.
IS. A method of detecting microbial pathogens in a body fluid comprising:
a. providing an aqueous filter solution between first and second injectable zones within a centrifugation tube, said aqueous filter solution containing a thermally sensitive gelling agent and being an aqueous solution of material which is non-toxic to said microbial pathogens and has a greater density than said sample fluid but able to selectively receive microbial pathogens from said sample fluid;
b. positioning said centrifugation tube such that said aqueous filter solution rests on said first injeetable zone and thereafter cooling said centrifugation tube to cause said thermally sensitive gelling agent to solidify said aqueous filter solution;
c. depositing a treatment fluid for said body fluid upon said solidified aqueous filter solution by passing an injection needle through said second injectable zone and thereafter injecting said sample treatment fluid into said centrifugation tube and upon said solidified aqueous filter solution:
d. depositing a sample of said body fluid upon said solidified aqueous filter solution and admixing said body fluid with said treatment fluid by passing a hypodermic needle through said first injectable zone and through said solidified aqueous filter solution and thereafter injecting said body fluid through said hypodermic needle into said centrifugation tube;
e. heating said eentrifugation tube sufficiently to liquefy said thermally sensitive gelling agent and thereby liquefy an aqueous filter solution;
f. subjecting said eentrifugation tube to centrifugation in a manner to force said sample of body fluid against said aqueous filter solution and cause said microbial pathogens to selectively pass therein and thereby separate from the residual mass of said body fluid sample and said treating fluid; and
g. separating said aqueous filter solution containing said microbial pathogen from said residual mass of said body fluid.
16. The method of claim 15 wherein said body fluid is blood and said treatment fluid comprises a blood lysing agent.
17. The method of claim 16 wherein said aqueous filter solution comprises an aqueous solution of a non ionic sugar.
18. The method of claim 17 wherein said nonionie sugar is sucrose.
19. The method ofclaim 17 wherein said aqueous filter solution contains at least about 40 wt of said sugar therewithin.
20. The method of claim 16 wherein said aqueous fil- 21. The method of claim 20 wherein said aqueous filter solution comprises an aqueous solution of a copolymer of sucrose and epichlorohydrin which has a molecular weight in the range of from about 300,000 to 500,000 and a specific rotation [01],, of 56.6.
22. The method of claim 21 wherein said aqueous solution contains from about 10m about 40 weight percent of said copolymer.
23. The method of claim 20 wherein said aqueous filter solution comprises an aqueous solution of dextran which has a molecular weight from about l0,000 to about 2,000,000.
24. The method of claim 23 wherein said dextran is present in said aqueous solution in an amount ranging from about 10 to about 40 weight percent thereof.
25. The method of claim 16 further comprises thoroughly mixing said aqueous filter solution containing said microbial pathogens; plating at least portions of said admixed, aqueous filter solution on a growth media for said microbial pathogens.
@22 2 UNITED STATES PATENT o FICE CERTIFICATE OF CORRECTICN Patent No. 3,875,012 Dated P 19.75
Inventor{s) Gordon L. Born and Joseph M. Hill It is certified that error appears in the above-identified patent and that said Letters Patent are hereby corrected as shown below:
Column 4, line 27, "sufficient" should be -sufficiently"; line 43; "from about 50 wt should be --from about 40 to about 50 wt Column 6, line 17, "exmaple" should be -example-.
'Column 7, line 21, "for" should be from--.
2 0 Column 12, line 11 (Claim 21), "[d] of 56.6" shouldbe 20 "m1 of 56.6'.
Signed and sealed this lst day of July 1975.
(SEAL) Attest:
' C. MARSHALL DANN RUTH C MASON Commissioner of Patents Attesting Officer and Trademarks
|Cited Patent
|Filing date
|Publication date
|Feb 4, 1937
|May 3, 1938
|Cook Lab Inc
|Multiple compartment ampule and hypodermic syringe
|Nov 5, 1937
|May 23, 1939
|Cook Lab Inc
|Controllable transfer element for multiple ampules
|Apr 30, 1938
|Mar 12, 1940
|Cook Lab Inc
|Controllable transfer element for multiple compartment ampules
|Mar 23, 1949
|Apr 1, 1952
|Cwi Sommerstein
|Dec 30, 1960
|Jan 8, 1963
|Grace W R & Co
|Isolation of bacteria
|Citing Patent
|Filing date
|Publication date
|Dec 20, 1974
|Jan 13, 1976
|J. K. & Susie L. Wadley Research Institute And Blood Bank
|For isolating pathogenic microorganisms
|Mar 24, 1976
|Jul 26, 1977
|J. K. And Susie L. Wadley Research Institute And Blood Bank
|Sample mixing and centrifugation apparatus
|Jun 5, 1978
|Sep 18, 1979
|Beckman Instruments, Inc.
|Layering a liquid sample onto centrifuge tube liquid contents
|Oct 10, 1978
|Jun 24, 1980
|Owens-Illinois, Inc.
|Fluid collection apparatus
|May 1, 1978
|Jul 1, 1980
|Owens-Illinois, Inc.
|Fluid collection apparatus
|Oct 18, 1978
|Jul 15, 1980
|J. K. And Susie L. Wadley Research Institute And Blood Bank
|Apparatus for detecting microbial pathogens employing a cushioning agent
|Feb 17, 1981
|Nov 9, 1982
|Beckman Instruments, Inc.
|Penetrable centrifuge tube
|Apr 2, 1981
|Dec 21, 1982
|Becton, Dickinson And Company
|Contamination-free separation device
|May 22, 1981
|Nov 22, 1983
|Concord Laboratories, Inc.
|Sterilization indicator
|Nov 30, 1982
|May 7, 1985
|Abbott Laboratories
|Powder syringe mixing system
|Aug 30, 1982
|May 28, 1985
|Automatic Liquid Packaging, Inc.
|Container having pierceable insert
|Sep 24, 1980
|Sep 24, 1985
|Boehringer Mannheim Gmbh
|Process for detecting pathogens
|Sep 29, 1983
|Apr 8, 1986
|E. I. Du Pont De Nemours And Company
|Method for the rapid detection of virus and viral antigens
|Sep 30, 1983
|Jun 24, 1986
|Concord Laboratories, Inc.
|Sterilization indicator
|Oct 27, 1983
|Jul 22, 1986
|Abbott Laboratories
|Container mixing system with externally mounted drug container
|Feb 22, 1984
|Aug 19, 1986
|Abbott Laboratories
|Container mixing system with externally mounted drug container
|Feb 3, 1986
|Mar 31, 1987
|Metal Box P.L.C.
|Containers for use in detecting micro-organisms
|Oct 28, 1983
|May 19, 1987
|Becton, Dickinson And Company
|Microbial pathogen detecting system and process
|Nov 4, 1986
|Aug 16, 1988
|Kerforshungsanlage Julich
|Chamber for the treatment of cells in an electrical field
|Jun 12, 1986
|Nov 1, 1988
|Abbott Laboratories
|Container system with integral second substance storing and dispensing means
|Jul 31, 1986
|Nov 22, 1988
|Abbott Laboratories
|Container for mixture of materials
|Nov 7, 1986
|Feb 20, 1990
|Claude Ranoux
|Container for fertilization of human ovocytes in the absence of CO2 -enriched air
|Nov 9, 1988
|Apr 28, 1992
|Wadley Technologies, Inc.
|Specimen transport device containing a specimen stabilizing composition
|Mar 28, 1989
|Aug 4, 1992
|Claude Ranoux
|Container for fertilization of human ovocytes in the absence of CO2 -enriched air
|Jul 19, 1991
|Jan 12, 1993
|Farmitalia Carlo Erba S. R. L.
|Chemical reagent vial
|Jul 24, 1992
|Sep 20, 1994
|Nissho Corporation
|Drug vessel
|Jan 12, 1993
|Jun 27, 1995
|Kabushiki Kaisha Seitai Kagaku Kankyusho
|Method for radioactivity measurement, process for preparing sample and device therefor
|Mar 14, 1994
|Oct 3, 1995
|University De Montreal
|Microbial contamination test device
|Feb 27, 1995
|Mar 26, 1996
|Dorn; Gordon L.
|Method for improving quantitative recovery of microorganisms from specimens containing blood components
|Jun 30, 1994
|Oct 29, 1996
|Science Incorporated
|Fluid delivery apparatus
|Jun 17, 1996
|Jan 27, 1998
|Tecan U.S.
|Laboratory reactor apparatus
|May 13, 1996
|Apr 7, 1998
|Steris Corporation
|Self contained biological indicator
|Feb 22, 1996
|Apr 28, 1998
|Nikon Corporation
|Case for housing a substrate
|Aug 2, 2004
|Dec 20, 2005
|Kirvan Clifford J
|Method and apparatus for pressurizing plastic pipe
|Sep 14, 2004
|Apr 20, 2010
|Goldenberg Alec S
|Controlled additive/reactant delivery system
|Jul 29, 2011
|Feb 24, 2015
|Medinstill Development Llc
|Sealed containers and methods of filling and resealing same
|May 28, 2010
|Jun 9, 2015
|Medinstill Development Llc
|Resealable containers and methods of making, filling and resealing same
|Dec 4, 2002
|Aug 21, 2003
|Dave Smith
|Flow immunoassay scanning assembly
|Dec 4, 2002
|Aug 21, 2003
|Dave Smith
|Plunger-based flow immunoassay assembly
|Dec 4, 2002
|Sep 4, 2003
|Dave Smith
|Drug and alcohol assay assembly
|Aug 13, 2003
|Feb 19, 2004
|Dewalch Binz
|Method and apparatus for processing substances in a single container
|Jul 21, 2004
|Jan 26, 2006
|Wong Cai Ne W
|Universal tissue homogenizer device and methods
|Sep 14, 2004
|Mar 16, 2006
|Goldenberg Alec S
|Controlled additive/reactant delivery system
|Jan 2, 2008
|Oct 23, 2008
|Dewalch Norman Binz
|Method and apparatus for processing substances in a single container
|Nov 17, 2011
|Py Daniel C
|Sealed Containers and Methods of Filling and Resealing Same
|Apr 21, 1977
|Nov 2, 1978
|Wadley Res Inst & Blood Bank
|Sample mixing and centrifugation appts. - for detecting microbial pathogens
|Oct 22, 1977
|May 18, 1978
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|Verfahren und vorrichtung zur bestimmung pathogener mikroorganismen
|Jul 8, 1987
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|Andreas Szabados
|Filtrationseinheit, insbesondere fuer medizinische proben
|Mar 26, 1992
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|Einrichtung zur sicheren entnahme von blut aus einem vorratsgefaess
|Mar 13, 1997
|May 7, 1997
|Macherey Nagel & Co Chem
|Trenneinrichtung zur Trennung von Substanzen
|U.S. Classification
|International Classification
|Cooperative Classification
|European Classification


text:
Volatility has been on the rise in global fixed income markets this year. Investors have grown increasingly cautious heading into the first Fed tightening cycle in nearly a decade. However, there are still pockets of value to be found in this highly uncertain environment.
Read More
Having been relatively calm for most of the summer, the markets were hit by a surge of volatility in August. However, the changes taking place in the markets are bringing opportunities as well as challenges.
Read More
China's surprise currency devaluation sparked declines in global equity markets and heightened concerns about China's economy. While markets may remain volatile, China and the Asia region are not facing financial crisis, and the sell-off has created attractive opportunities.
Read More
Bridge the generational gap and win with plan sponsors.
Millennials are here
With the increased focus on tax-aware investing, a trend of accumulation only products and platforms is emerging in the broader VA competitive landscape.
Campaign Overview
T. Rowe Price Chairman & CIO Brian Rogers
The Current Views of the Asset Allocation Committee
Our Products, Differentiators, and Evaluation Process

subset: commoncrawl


text:
Do You Find Yourself More Attractive in Photos or In Person?
As for me, I don't know. I guess I would need an honest second opinion since I've never actually seen myself in person besides in the mirror. Right now though, I'd have to say I look better in photos than in person.
No clue, I'd say I probably look like crap in both forms! ;)
I'd say in person, I take really bad pics, except for a few that dh did.
I am not photogenic at all....I hate pictures of myself...lol...
I think I look better in person:)
Quoting luvmybug:
FYI i would like to actually MEET everyone in this group so we could find an answer. you ladies rock! muahs!
I definitely look better in person. I take sucky pictures. In your picture above you sort of remind me of Snookie (lol). Your face is thinner than hers though.


text:
The temperature outside is much higher than I'd like it to be and, as it's only July, we have a lot more warm weather to look forward to this year (especially in Southern California, if last summer's heat was any indication!). Cold desserts are a great way to cool down when it's hot out. One of my favorites is the ice cream sandwich, because it ... Read More »
Category Archives: Sweet Stuff
Fake cakes for weddings
Wedding cakes do not come cheap. In fact, the Bridal Association of America says that the average wedding cake costs $543. And the more elaborate you want your cake to be, the more people you intend to serve, the more it is going to cost you. One way to cut corners and reduce food costs is to cut down on the quality of the ingredients ... Read More »
Starbucks to release new chocolates, hot chocolate
Starbucks and Hershey's have recently come togetherto put out a new line of coffee-flavored, Starbucks branded chocolates, which should be available in stores in just a few months. Starbucks currently sells a line of chocolates in their stores, but these new items will be sold at grocery stores, much like the bottled frappuccinos and coffee drinks are now. The companies are planning a lineup that ... Read More »
Pimp that Animal Cookie!
Blame it on the "super size me" world that we live in, but I'm a big fan of the site Pimp That Snack, where users create their own giagantic – a.k.a. pimped – versions of popular snack foods. Top ranked projects include the Creme de la Creme Egg, Giant Twix, Rubiks Cake and a McDonalds Apple Pie. Since I first started visiting the site, I've ... Read More »
Cupcakes for sale, free car with purchase
Everyone likes to get something for free. The idea that you're getting a little something extra can really sweeten the deal when you're making a purchase. A, who blogs at Never Bashful With Butter, took the idea of sweetening a deal a step further and gave away a batch of her impressive homemade cupcakes to the person who purchased her car. Or, if you want ... Read More »
Multigrain donuts from Krispy Kreme
Bread or pastry made with whole grains has an advantage over those made with plain white flour because they have more heart-healthy fiber in the finished product. If you eat a lot of bread, switching to a whole grain version can have a positive impact on your diet. If you only eat a sandwich once in a while, it doesn't make that much difference. And ... Read More »
How to keep brown sugar soft
Brown sugar can be a bit problematic because, when left unattended, the moist sugar can quickly transform into a rock-like state. This happens because the sugar dries out and the molasses used to keep it brown (most brown sugars are white sugars, with molasses added back) hardens and sticks it together like glue. There are several ways to avoid this problem. The easiest is by storing the ... Read More »
"Light" whipped cream isn't that light
Whipped cream is one of the more waistline-unfriendly foods you can eat. It's high in fat, can be high in sugar (and thus, high in calories) if over-generously sweetened and it is really hard to limit yourself to a two tablespoon serving size when slices of apple pie, ice cream sundaes and molten chocolate cakes call out for more than just one mouthful – and ... Read More »


text:
- Sort:
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Manhattan Beach, pier, California, Christmas, lights, twilight, view north-south, Catalina island, Palos Verdes, Santa Monica, 200 degrees, Nikon P5100, 12mp, 31 images. I forgot to put lens on telephoto and shot only one column x 31 shots wide. Considering the extreme lighting differences the image came out OK. ©...
Christmas decorated house in Roanoke, Virginia on Westover Avenue. Photo notes: Tungsten white balance, camera was on a 6 second self timer to eliminate any vibration from the GigaPan imager then went to a two second exposure at 80 ISO for the clearest resolution. I also programed a 15 second wait time between exposu...
Festival of Lights December 2008 marked the 12th Annual Festival of Lights inside Kauai's Historic County Building, a celebration created by the community to continue a local tradition and preserve the glorious collection of Folk Art created by Auntie Josie...
This is my first night GigaPan that I have been wanting to take. Its of one of the places in Hawaii that you can get the best view of the Honolulu-Waikiki city lights. Known by all locals and many visitors. これは、私がずっと撮りたかった初めての夜のGigapanです。これは、ハワイ州の一�...
This is a higer resolution image of the lights of Casa del Pomba - The House of the Dove - in Livermore California, the hub of Photon Valley. This annual event uses 301,000 lights to transform Deacon Dave's front yard into a very special place. Hundreds of laughing children and smiling adults come through here each ...


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Replacement frame lock lever, including nut.
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Publications : Browse
Browse PATH publications
Subject: Reproductive health > Family planning
Publication date: All
- The Case for Menstrual Health: PATH Works to Build Awareness, Commitment, and Access to Improved Menstrual Care
This fact sheet describes PATH's work with partners and countries to address and improve menstrual health in low-resource settings by identifying affordable and appropriate menstrual care products, increasing education and awareness, and advocating for further investment.
Publication date: November 2015
Region: Global
- Monitoring Sayana® Press Pilot Introduction
In July 2014, PATH launched a coordinated introduction of the subcutaneous injectable contraceptive Sayana Press in Burkina Faso, Niger, Senegal, and Uganda. This brief features monitoring results and lessons learned to date, which can help inform contraceptive introduction programming for Sayana Press and other methods.
Publication date: October 2015
- The Case for Investing in Research to Increase Access to and Use of Contraception Among Adolescents
This 80-page report highlights why governments and donors should invest now in research to help determine and implement the most effective and efficient ways to enable adolescents to access and use contraception. The long-term economic and health benefits of increasing access will far outweigh the costs.
Author: Greene M.E., Merrick T
Publication date: August 2015
Region: Global
- Countering Myths and Misperceptions About Contraceptives
This issue of Outlook examines the scope and impact of misperceptions about contraception, the science behind debunking misperceptions, and approaches that programs are using to address this persistent problem.
Author: Wells E
Publication date: June 2015
Region: Global
Part of series: Outlook
- SILCS Diaphragm: A Reusable System for Microbicide Gel Delivery
The SILCS single-size diaphragm was designed to expand women's options for nonhormonal contraception. The SILCS diaphragm is also being assessed as a low-cost, reusable option for microbicide gel delivery and provide contraceptive protection at the same time.
Publication date: June 2015
Region: Global
- SILCS Diaphragm: Summary of Clinical Evaluations
The SILCS diaphragm has been evaluated in ten studies conducted in multiple countries since 1998. These studies include design validation and consumer use studies, as well as clinical studies of safety, effectiveness, and acceptability. This document provides summaries of the key studies implemented during development and validation of SILCS as a barrier contraceptive.
Publication date: June 2015
Region: Global
- SILCS Launches in Early Markets
The SILCS diaphragm is now available in over 20 countries, including Europe, Canada, Australia, Malaysia, and the United States. PATH led a user-centered design process involving input from women, their partners, and providers whose feedback resulted in its innovative design that is easy and comfortable to use.
Publication date: June 2015
Region: Global
- Policy Brief on the Case for Investing in Research to Increase Access to and use of Contraception Among Adolescents
This document outlines why governments and donors should invest now in research to help determine and implement the most effective and efficient ways to enable adolescents to access and use contraception. It summarizes the findings of a longer technical report.
Publication date: March 2015
Region: Global
- Sayana® Press Project
These fact sheets describe PATH's pilot introduction project to deliver the injectable contraceptive Sayana® Press (depot medroxyprogesterone acetate or DMPA) to women in sub-Saharan Africa, as well as research on the potential for self-injection of Sayana® Press. The fact sheets also provide clinical information about Sayana® Press.
Publication date: January 2015
- Reproductive Health Global Program Website
This website about PATH's Reproductive Health global program provides an overview of the program and its featured projects. See other PATH program websites.
Publication date: 2015
Region: Global
Part of series: Program websites
- Reproductive Health Supplies Coalition Website
The Reproductive Health Supplies Coalition is a global partnership of public, private, and nongovernmental organizations dedicated to ensuring that all people in low- and middle-income countries can access and use affordable, high-quality supplies to ensure their better reproductive health.
Publication date: 2015
Region: Global
Part of series: Websites
- Vietnam Country Program Website
This website about PATH's Vietnam country program provides an overview of the program and its featured projects. See other PATH program websites.
Publication date: 2015
Region: Asia
Part of series: Program websites
- PATH in China: Joining With Diverse Partners to Create Sustainable Health Solutions
This brochure outlines PATH's work in China, with emphasis on public-private partnerships for product development. Areas of focus include vaccines and immunization, health technologies, and women's health.
Publication date: September 2014
Region: Asia
Part of series: Country brochures
- Integrating Female Condoms Into HIV Prevention Programs: A Case Study of Barriers, Facilitators, and Future Opportunities in Kenya
This report examines the factors that have supported and hindered the inclusion of female condoms in two HIV prevention programs in Kenya—prevention of mother-to-child transmission of HIV and voluntary medical male circumcision—and offers potential actions to help improve integration and access in future programming.
Author: Whipkey KJ, Coffey PS, East L, Omurwa T, Murunga P
Publication date: August 2014
- Policy Advocacy for Total Market Approaches for Family Planning: Eastern Europe and Central Asia Regional Workshop May 2014
In May 2014, PATH and the United Nations Population Fund convened a regional workshop on policy advocacy for total market approaches (TMAs) for family planning, in Antalya, Turkey. Seventy-three participants from 18 countries gathered to learn more about advocacy strategy development and to plan advocacy activities to bring about policy changes in support of their TMA action plans.
Publication date: August 2014
Region: Eastern Europe, Global
- Communications Guidance for Introducing Sayana® Press
This guide was created to support ministries of health and nongovernmental implementing partners as they develop communications strategies related to the introduction of the injectable contraceptive Sayana® Press. The document outlines communication strategies, audiences, key messages, and communications channels recommended for successful introduction of the product. For more information about the Sayana® Press Introduction and Evaluation project, visit http://sites.path.org/rh/?p=292.
Author: Lasher H
Publication date: February 2014
Region: Africa
- Addressing Adolescent Sexual and Reproductive Health in a Complex World: Outlook 30th Anniversary Issue
Adolescents and their sexual and reproductive health needs are in the global health spotlight. This issue of Outlook provides an overview of the sexual and reproductive health needs of adolescents, gives examples of different approaches to addressing those needs, and highlights some of the most intractable challenges.
Author: Wells E
Publication date: December 2013
Region: Global
Part of series: Outlook
- Improving Women's and Children's Health in Senegal
This fact sheet provides an overview of PATH's work in Senegal. Since 2001, PATH has worked side by side with government and community partners in Senegal to reduce the burden of infectious disease, improve family planning and reproductive health, and strengthen health systems to save lives.
Publication date: December 2013
Region: Africa
- Sayana® Press Training Materials
These training materials focus on the administration of the injectable contraceptive Sayana® Press for facility- and community-based providers and were developed as part of the Sayana® Press pilot introduction project. For the original Word and PowerPoint files to customize for your training program, please email firstname.lastname@example.com.
Publication date: November 2013
Region: Global
- Improving Reproductive Health Through Innovation: PATH Increases Access to Products, Services, and Information
This fact sheet highlights PATH's work to address family planning needs, provide better methods of protection against sexually transmitted infections and HIV, and improve diagnosis and treatment of women's cancers in low-resource settings.
Publication date: October 2013
Region: Global
Part of series: Areas of focus fact sheets


text:
7. Do make mealtime family time.
There is evidence that eating with children and having regular family meals helps prevent childhood obesity, according to James Mitchell, MD, a psychiatrist and eating disorders expert at Sanford Health in Fargo, N.D.
"Research shows that when you eat with your child, they eat more slowly and make healthier choices," Mitchell says. "When you have family meals, eating becomes a social event."
8. Don't make kids follow a strict exercise plan.
Torres suggests that parents encourage physical activity as a natural part of life, not a chore, or kids are likely to resist. "Make it positive," she says. "Instead of making your kid go to a cardio class once a week, for example, do something spontaneous, like take a walk around the neighborhood. Set goals to encourage your child, such as, 'Let's see if we can walk a little further this time.'"
9. Do make sure your child is getting enough sleep and is 'unplugged.'
More and more studies are being released that link a lack of sleep to weight gain and other medical illnesses. Work to make sure that your child is getting enough sleep every night.
Sleep needs vary from child to child, but the following are general guidelines from the National Sleep Foundation:
- 1 to 3 years old: 12 to 14 hours per day
- 3 to 5-year-olds: 11 to 13 hours per day
- 5 to 12-year-olds: 10 to 11 hours per day
- 12 to 18-year-olds: at least 8 1/2 hours per day
To help your child get the right amount of Zzzs, remind him to 'unplug' from the computer, cell phone, and TV at least two hours before bedtime. Artificial light from electronics stimulates the brain and may make it harder to fall asleep.
Also, remember, the more time your child spends watching TV or being on the computer or phone, the less time he's spending having positive interactions with family or being physically active.
10. Do make it clear that you love your child unconditionally.
Remember that your long-term goal as a parent is the same whether your child is size 4 or 14: to raise a person who is comfortable with herself and knows that she is loved.
"Be careful of the messages you send," Stone says. "You never want your child to believe that your love for her is based on what she eats or doesn't eat."
As a once overweight child, Torres agrees. "Kids need to know that what you feel about them has nothing to do with their weight," she says. "Part of loving yourself means taking care of your body and keeping it healthy. If your child knows she's loved and learns to love herself, she's far more apt to make healthy choices."


text:
Don't Cry For Me, Argentina
When it comes to political sex scandals, we talk a great deal around here about what's just plain prurient and what's not. Granted, when something like Governors Spitzer and/or Sanford breaks, it is a news story first and foremost — that's what they sign up for when they take the oath (not the "I do" promise, natch, but the other one). So when I saw the text of the cringingly intimate emails between the Governor of South Carolina and the (apparently) beautiful Maria — I was pretty uncomfortable. (Um, yes, I read them all the way through. I'm HUMAN.) But I wondered if that kind of private language adds anything to the discussion of his performance as Governor, or if splashing it all over The State just adds more fuel to a fire that's kinda... mean. Who among us has not spoken of their "apocalyptic passion" in a letter or email, and would pretty much DIE of embarrassment if anyone — other then our beloved* — ever read it? Listen, Sanford screwed up, for sure, and he is taking a mighty punishment right now — the question is, do we need to rub salt in those
tan lineswounds, as well?
*Dear [redacted], please burn that letter that I sent you in the summer of '95. Hope all is well with you. Best, Barrie
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text:
George William Roberts
Position: P
Height: 6-0 Weight: 181 lbs.
Born: June 10, 1955 in Lynchburg, VA
High School: E.C. Glass
College: Virginia Tech (school history) (Roberts college stats)
Weighted Career AV (100-95-...): 10 (6993rd overall since 1950)
Other SR Links: College Football at Sports-Reference.com
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Output includes all games from 1920 to 2015.


text:
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20 apartments for rent near Nowell Pointe, Raleigh, NC
Apartments in Nowell Pointe, Raleigh, NC
Raleigh's Nowell Pointe neighborhood borders North Carolina State (N.C. State) University. In fact, Nowell Pointe is within only a mile of the sprawling Wolfpack campus, located to the southeast of the community.
The Nowell Pointe neighborhood is flanked by its namesake, Nowell Pointe Drive, as well as Cedar Down Drive, Nowell Ridge Road, Nowell Road, and Cedar Waters Drive. Located in the 27607 ZIP code, the community is conveniently close to Chapel Hill Road and the busy Hillsborough Street. Nearby are two major commuting highways, I-40 and I-440.
The community is filled with mainly two-story colonial homes with two to three baths and approximately 2,500 square feet of space. Most residences were built in the late 1990s. Nowell Pointe is bordered by the neighboring communities of Ashbury and Westover. Fortunately, there are plenty of trees and woods lining the neighborhood to preserve green and open spaces.
Commuting to work and traveling are both convenient for Nowell Pointe residents. The community is just miles from Raleigh-Durham International Airport; ECPI Technical College; and the RBC Center, which is home to the Carolina Hurricanes ice hockey team. Carter Finley Stadium is also readily accessible to Nowell Pointe.
Raleigh enjoys rapid population growth and high residential turnover, largely due to the area's plentiful high-tech employment opportunities in Research Triangle Park and surrounding commercial centers. As a result, the median household age of many local communities, including Nowell Pointe, is only 32. Many residents are singles or young married couples.
Nearby schools include a diverse mix of public, private, and parochial institutions. Within a few miles of the neighborhood are the Al Iman private school, Bridges Alternative public school, Cardinal Gibbons High School, Follow the Child Montessori private school, and Fred Olds Magnet School.
- Olde Raleigh
- Cameron Village
- Trinity Square
- Oakdale
- Bashford Ridge
- Asbury Park
- MT Vernon Park
- Brandywine
- Meadow Springs
- Bradford Place


text:
Booktopia Comments
Order anything from our Christmas Gift Guide and you could win a Christmas Prize Pack valued at $1000! *Terms and Conditions apply.
Product Description
In the early 1930s, Nancy Wake was a young woman enjoying a bohemian life in Paris. By the end of the Second World War, she was the Gestapo's most wanted person.
As a naïve, young journalist, Nancy Wake witnessed a horrific scene of Nazi violence in a Viennese street. From that moment, she declared that she would do everything in her power to rid Europe of the Nazis. What began as a courier job here and there became a highly successful escape network for Allied soldiers, perfectly camouflaged by Nancy's high-society life in Marseille. Her network was soon so successful - and so notorious - that she was forced to flee France to escape the Gestapo, who had dubbed her "the white mouse" for her knack of slipping through its traps.
But Nancy was a passionate enemy of the Nazis and refused to stay away. Supplying weapons and training members of a powerful underground fighting force, organising Allied parachute drops, cycling four hundred kilometres across a mountain range to find a new transmitting radio - nothing seemed too difficult in her fight against the Nazis.
Peter FitzSimons reveals Nancy Wake's compelling story, a tale of an ordinary woman doing extraordinary things.
About the Author
Peter FitzSimons is a journalist with the Sydney Morning Herald and Sun-Herald. He is the author of nearly twenty books - including Tobruk and biographies of Nancy Wake, Kim Beazley, Nene King, Nick Farr-Jones, Steve Waugh and John Eales.
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Comments about Nancy Wake Biography Revised Edition:
All our younger generation should read this book and marvel at what a true hero this wonderful person is
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Comments about Nancy Wake Biography Revised Edition:
Easy read.
Best Uses
Comments about Nancy Wake Biography Revised Edition:
Written with insight and engaging from the first chapter. Peter Fitzsimons brings this person to life. A book that should be adaptable to all ages
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Comments about Nancy Wake Biography Revised Edition:
This book was fantastic, I would recommend, that everyone should read it.
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Comments about Nancy Wake Biography Revised Edition:
Easy and informative
Audience: General
Format: Paperback
Language: English
Number Of Pages: 416
Published: 8th November 2011
Publisher: HarperCollins Publishers (Australia) Pty Ltd
Country of Publication: AU
Dimensions (cm): 19.6 x 12.7 x 2.6
Weight (kg): 0.3


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A Shrove Tuesday spaghetti dinner will be served today at St. Paul the Apostle Catholic Church, 125 S. Spruce St., Annville.
The event will be held in the social hall from 1 to 4 p.m. and 4 to 7 p.m. The dinner will consist of spaghetti and meatballs, salad, bread, a beverage and dessert and will cost $8 for a large meal and $5.50 for a small meal.
Proceeds will benefit parish needs and be used to fund ongoing projects in Third World countries.
St. James Lutheran Church in Lebanon will host a men's breakfast at 8 a.m. Thursday at the Cedar Grill and a Bible study at 4 p.m. that same day. The fellowship circle will have its cleanup day Saturday at 9 a.m.
For more information, call the church at 717-272-5563 between 9 a.m. and 1 p.m.
St. Matthews Lutheran Church, 534 E. Lehman St., Lebanon, will hold an Ash Wednesday service at 7 p.m. Wednesday. A soup and sandwich supper will be held in the church at 5:30 p.m.
A women's prayer breakfast will be held at 8 a.m. Ash Wednesday at Heisey's Diner in North Lebanon Township. The group will continue to meet each Wednesday through Lent at the same time and location and will study the theme "The Gift That Keeps Giving."


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The logo of the social networking website Twitter displayed on a computer screen in London. / Leon Neal, AFP/Getty Images
SAN FRANCISCO - Twitter is celebrating its 8th birthday by taking millions of its users back to their first tweets.
The microblogging service on Thursday launched a website, called first-tweets.com, that lets users search their own account or those of others to find their first tweet. Users are also encouraged to use the hashtag #FirstTweet.
"Millions of prolific tweeters have made Twitter an exciting, fun and powerful place to connect with others," says Gabriel Stricker, Twitter's vice president of marketing and communications. "But each of you had to start somewhere."
If anything, Twitter celebrated its birthday early - in November - when it went public in the most important technology offering since Facebook's market debut in May 2012. Twitter has become an essential tool for celebrities, politicians and citizens to share information, opine and vent.
Still, questions remain over Twitter's financial prospects and how the stock will perform once subject to quarterly earnings reports.
Of course, no first tweet was more notable than the one posted by Twitter co-founder Jack Dorsey on March 21, 2006, which happened to be the first tweet ever published. "Just setting up my twtr," Dorsey wrote eight years ago. Ironically, Dorsey has been noticeably quiet on Twitter the past few months.
Dorsey, a computer programmer from St. Louis who is Twitter's chairman, considers Twitter a "status updater, a way to say where he was and what he was doing. A place to display yourself, your ego."
Twitter co-founder Evan "Ev" Williams, a computer-obsessed Nebraska farm boy, has a different opinion. Twitter is a way to learn "where other people were and what other people were doing," he says.
Here's a look at some other first tweets, including from USA TODAY's Tech section. Twitter has some more first tweets on its blog.
Twitter has grown quickly in the last eight years, morphing from a simple social network to a public company, making its debut on the New York Stock Exchange late last year. As of last month, Twitter boasted more than 240 million monthly active users.
"Twitter changed my personal and professional life," says Bijan Sabet, a general partner at Spark Capital, which led a $14 million Series B round of funding for Twitter in 2008 when it had about a dozen employees.
"In many ways, Twitter has innovated, evolved and grown - but it is still the very same product I fell in love with from the beginning," Sabet says. "It's a simple, powerful way to connect with the world in real time."
Follow Brett Molina on Twitter: @bam923.
Copyright 2015 USATODAY.com
Read the original story: As Twitter turns 8, here's how to find your first tweet


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Destiny trilogy - speculations
View Single Post
March 4 2008, 05:28 AM
William Leisner
Location: Minneapolis, MN
Re: Destiny trilogy - pretty heavy spoilers
David Mack wrote:
JoeZhang wrote:
I found it here:
Just to put things in perspective for everyone, BTW, amazon.co.uk didn't even get the book's
right on their page.
I'm just saying.
This made me curious, so I clicked on the Amazon UK link for
Infinity's Prism
Building on the concept behind the Mirror Universe novels, Myriad Universes takes the theme a step further, into a world where instead of one alternate universe with mirror versions of many Star Trek characters, one single character is reflected across three radically different Star Trek universes. Thus through the very different visions of three separate authors readers can discover three entirely new facets of Captain Jean-Luc Picard. Follow our favourite captain into a universe of adventure -- where history takes a different turn and the only limit to what can happen is imagination.
Let me say right here that this is utter and complete baloney. I don't know about Christopher's or James's stories, but Jean-Luc Picard does
in any way, shape or form appear in
A Less Perfect Union
. If the description of
is anywhere near as accurate, then I almost have to wonder if it actually is a
Star Trek
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