Difference between revisions of "Phage therapy"

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==Contribute To Our Pilot Program!==
If you have time or resources to contribute to this project, we urge you to attend our bi-weekly meetings or donate to our Pilot Program WePay Campaign [https://wepay.com/donations/gonorrhea-eradication-team here!]
==The Gonorrhea Eradication Team and Integration Taskforce (GETit!) ==
We are a volunteer group of experienced citizen scientists working to foster healthy communities through innovative medical research. Our team includes PhD-, Master- and Bachelor-level scientists. Together, we are taking on our first major project: the eradication of antibiotic resistant gonorrhea using Phage Therapy! We are asking for support in launching our Pilot Program which aims to: (1) Isolate and Characterize Bacteriophage from 10 patients, and (2) create a proof-of-concept bacteriophage cocktail to be tested ''in vitro''.
''Our team:''
:'''Craig Rouskey''', MSc in Molecular Biology, Biochemistry, Microbiology and Immunology
:'''Ami Knop Ullrich''', MSc in Medical Microbiology and Immunology
== Introduction ==
== Introduction ==
: ''Neisseria gonorrhoeae'' has progressively developed resistance to the antibiotic drugs used to treat it. Since the late 1970's, Gonorrhea has shown signs of developing resistance to 3rd-generation, cephalosporin antibiotics which are ultimately the last line of defense against this bacterial pathogen <ref>http://www.cdc.gov/std/Gonorrhea/arg/default.htm</ref>. Antibiotic resistant Gonorrhea is therefore a growing public health concern. In the United States, this concern is exacerbated by the fact that primary treatments for gonorrheal infections are solely antibiotic-based. Currently, CDC STD treatment guidelines recommend dual therapy with the injectable cephalosporin ceftriaxone and either azithromycin or doxycycline to treat all uncomplicated gonococcal infections among adults and adolescents in the United States. Dual therapy is recommended to address the potential emergence of gonococcal cephalosporin resistance. Given the ability of ''N. gonorrhoeae'' to develop antibiotic resistance, it is critical to continuously monitor gonococcal antibiotic resistance and encourage research and development of new treatment regimens for gonorrhea <ref>http://www.cdc.gov/std/Gonorrhea/arg/basic.htm</ref>.
Neisseria gonorrhoeae, the sexually transmitted pathogen that causes Gonorrhea, has progressively developed resistance to the antibiotic drugs used to treat it. Within the past 40 years, this pathogen has developed resistance to 3rd-generation, cephalosporin antibiotics which are ultimately the last line of defense against this bacterial pathogen <ref>http://www.cdc.gov/std/Gonorrhea/arg/default.htm</ref>. Antibiotic resistant Neisseria gonorrhoeae (ARNG) is therefore a growing public health concern. In the United States, this concern is exacerbated by the fact that primary treatments for gonorrheal infections are solely antibiotic-based. Currently, CDC STD treatment guidelines recommend dual therapy with the injectable cephalosporin ceftriaxone and either azithromycin or doxycycline to treat all uncomplicated gonococcal infections among adults and adolescents in the United States. These treatments have been mostly successful, but given the ability of ARNG to develop antibiotic resistance, it is critical to continuously research and develop new treatment regimens for gonorrhea<ref>http://www.cdc.gov/std/Gonorrhea/arg/basic.htm</ref>.  


: This project was established to (1) Provide surveillance insight into the emergence of antibiotic resistant gonorrhea, (2) better understand and characterize bacteriophages from antibiotic-resistant, sexually-transmitted, ''Neisseria gonorrhoeae,'' and (3) develop phage-based prophylaxis and therapy to stop prevent further emergence of antibiotic-resistant gonorrhea.
To meet the needs of a rapidly changing public health field in which traditional antibiotic-based therapies are becoming ineffective, the Gonorrhea Eradication Team and Integration Task-force (GETit) was created. This organization was created to conduct research into bacteriophage associated with Neisseria gonorrhoeae. Bacteriophage are viruses that infect bacterial hosts and can induce lysis upon infection. Ultimately, our goal is to identify and characterize bacteriophage from wild type ARNG and use a composite cocktail of ARNG-associated phage as therapy to treat hosts infected with this pathogen.
:<br />
 
: This project is in its early stages. We are working to:
'''During our pilot program, we plan to''':
# isolate ''N. gonorrhoeae'' from 10 patients
# isolate bacteriophage from 10 clinical samples of ARNG
# characterize bacteriophages from antibiotic-resistant, sexually-transmitted, ''Neisseria gonorrhoeae'', and  
# conduct proof-of-concept experiments in which ARNG-derived phage cocktails are used to lyse cultures of ARNG ''in vitro''.
:
<br />
: '''This project is in its early stages. Our longer-term goals are to''':
# Begin a free-Gonorrhea screening program on the streets of Oakland and San Francisco;
# Begin a free-Gonorrhea screening program on the streets of Oakland and San Francisco;
# Enhance the Center for Disease Control's Surveillance of antibiotic-resistant gonorrheal infections;
# Enhance the Center for Disease Control's Surveillance of antibiotic-resistant gonorrheal infections;
# Identify the first known bacteriophages from ''N. gonorrhoeae;''
# Develop assays for the rapid detection of ''N. gonorrhoeae.''
# Develop methods by which bacteriophages can be cultured and used as prophylaxis and therapy for gonoccocal infections;
# Develop methods by which bacteriophages can be cultured and used as prophylaxis and therapy for gonoccocal infections;
# Develop a space that has the equipment (acquired or built) we need in which we can conduct our research;
# Develop a space that has the equipment (acquired or built) we need in which we can conduct our research;
# Develop a community education program to bring science to underrepresented cultural groups.


:
:
: <br />
: <br />


:'''''If you would like to help with any of these project goals, please contact Craig (moleculararts [at] riseup [dot] net).'''''
:'''''If you would like to help with any of these project goals, please contact Craig (contact [at] craigrouskey [dot] com).'''''


== What is Phage Therapy ==
== What is Phage Therapy ==
: Phage therapy is a biological therapy that uses bacteriophages (bacterial viruses) to infect and lyse bacterial pathogens.<ref>[http://www.landesbioscience.com/journals/bacteriophage/AbedonBP1-2.pdf] Phage Treatment of Human Infections. Abedon, et al. 2011.</ref>  
: Phage therapy is a biological therapy that uses bacteriophages (bacterial viruses) to infect and lyse bacterial pathogens.<ref>[http://www.landesbioscience.com/journals/bacteriophage/AbedonBP1-2.pdf] Phage Treatment of Human Infections. Abedon, et al. 2011.</ref>  


== Sexually Transmitted Bacterial Infections ==
== Sexually Transmitted Bacterial Infections ==  
: This project will begin by studying ''Neisseria gonorrhoeae'' the causative agent of gonorrhea.
: Neisseria gonorrhoeae is a facultative intracellular pathogen that is able to infect the eye, pharynx, anus/rectum, urogenital tract, and may be disseminated throughout the body in more complex cases. The Center for Disease Control reports that in 2011 there were an 321,849 new cases of gonorrhea reported in the U.S.<ref>[http://www.cdc.gov/std/gonorrhea/STDFact-gonorrhea-detailed.htm] Detailed STD Facts </ref> of which about 50% are estimated to be reported ( for a total of 700,000 estimated new cases in 2011). The World Health Organization reports that there are between 65-105 million new cases of gonorrhea nationally each year. Of these, 0.5-3% of cases develop into disseminated, systemic infection where the facultative intracellular diplococci induce more serious illness such as pelvic inflammatory disease.
: Neisseria gonorrhoeae is a facultative intracellular pathogen that is able to infect the eye, pharynx, anus/rectum, urogenital tract, and may be disseminated throughout the body in more complex cases. The Center for Disease Control reports that in 2011 there were an 321,849 new cases of gonorrhea reported in the U.S.<ref>[http://www.cdc.gov/std/gonorrhea/STDFact-gonorrhea-detailed.htm] Detailed STD Facts </ref> of which about 50% are estimated to be reported ( for a total of 700,000 estimated new cases in 2011). The World Health Organization reports that there are between 65-105 million new cases of gonorrhea nationally each year. Of these, 0.5-3% of cases develop into disseminated, systemic infection where the falcutative intracellular diplococci induce more serious illness such as pelvic inflammatory disease.


== Symptoms of Gonorrhea ==
== Symptoms of Gonorrhea ==
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== History of Phage Therapy ==
== History of Phage Therapy ==
<br/>
:The initial discovery bacteriophage has been subject to speculation to who was the first. In the western world, Ernest Hankin, a British bacteriologist, first reported observing unidentified antibacterial preventing the spread of cholera (Vibrio cholerae) in the rivers Ganges and Jumna in India in 1896 [mini]. These unidentified antibacterial remained of unknown origin until bacteriologist Frederick Twort hypothesized that the cause of inhibition of bacterial growth was from viruses [M13] in 1915. Twort would be unable to continue pursuing his findings due to various reasons.  
:The discovery of bacteriophage have been subject to debate as to the offical claims of founding. Ernest Hankin, a British bacteriologist, first reported in 1896 on observation of an unidentified antibacterial preventing the spread of cholera (Vibrio cholerae) in the rivers Ganges and Jumna in India [21]. Russian bacteriologist Gamaleya observed a similar phenomenon while working with Bacillus subtilis [48]. The first to hypothesize Bacteriophage as a virus that infects bacteria host cells was Frederick Twort, bacteriologist from England in the early 1900s [70], however due to various reasons such as lack of funds, could not pursue his findings. It was not until two years later that Felix d'Herelle, a French-Canadian microbiologist at the Institut Pasteur in Paris "offically" discovers Bacteriophage, which he named after "bacteria" and "phagein" or to devour, in Greek[68,70].
 
<br/>
:D'Herelle first observed bacteriophages as 2-3mm "clear spots" which was a pathogenic agent to coccobacillus bacteria cultures studying locusts in South America and Africa in the early 1900s [M18]. He would use his observations of  bacteriophages to perform one of the first phage therapy techniques on severe hemorrhagic dysentery outbreaks among French soldiers stationed at Maisons-Laffitte in the summer of 1915 [M18, Mini]. On September 15, 1917 Felix d'Herelle would present his findings in the Academy of Sciences naming this phenomena 'Bacteriophage' after the Greek words "bacteria" and "phagein", which means to devour [M18]. 
:D'Herelle first observed bacteriophages studying microbiologic means of controlling an epizootic of locusts in Mexico in 1910. He later used his observations bacteriophages to perform one of the first phage therapy techniques known on severe hemorrhagic dysentery outbreaks among French soldiers stationed at Maisons-Laffitte in the summer of 1915. At Maisons-Laffitte, d'Herelle made bacterium-free filtrates of the patients' fecal samples and mixed and incubated them with Shigella strains isolated from the patients. A portion of the mixtures was inoculated into experimental animals (as part of d'Herelle's studies on developing a vaccine against bacterial dysentery), and a portion was spread on agar medium in order to observe the growth of the bacteria. It was on these agar cultures that d'Herelle observed the appearance of small, clear areas, which he initially called taches, then taches vierges, and, later, plaques [68]. His findings were presented in September 1917 meeting of the Academy of Sciences and later published [18].  
 
<br/>
:Continuing on the study of bacteriophages, d'Herelle starts researching on the effects of phage therapy on a 12-year-old boy with severe dysentery in 1919 at the Hôpital des Enfants-Malades in Paris, under the hospital's Chief of Pediatrics, Victor-Henri Hutinel. The patient's symptoms ceased after a single administration of d'Herelle's anti-dysentery phage, and the boy fully recovered within a few days [DHERELLE-BOOK]. Phage therapy was accept as the reason for cure after three more patients having bacterial dysentery were each treated with one dose of the preparation and started to recover within 24 hours of treatment.
:===Modern day Phage Therapy in Mammals===
 
<br/>
:Due to poor scientific experimentation, understanding of pathogenesis, phage-host interactions, and pharmacokinetics knowledge, bacteriophages soon lost interest in the western world as antibiotics and penicillin was discovered. However research continued in the eastern Europe in the Soviet era where two prominent research centers established, The Eliava Institute (EIBMV) in 1923 and The Hirszfeld Institute (HIIET) in 1952. Both continue to research bacteriophages presently. With new knowledge bacteriophages properties and due to increasingly drug resistant bacteria, bacteriophages has been brought to light again in the fight against disease.
:===Modern day Phage Therapy in Humans===
 
<br/>
 
== Present Day Phage Therapy in Animals ==
 
:Present day research on phage therapy in gram negative bacteria in animals have been very extensive and promising.
 
:With a better understanding of phage therapy and more advanced equipment, modern day phage therapy can close the gaps in experimental accuracy and knowledge that was missing in the past. Using blood culture machines and better filtration methods, such as caesium chloride density centrifugation, have lead the better understanding of host-phage relationships such as longer-circulating strains of phage [nih11]. This has led to more efficient phage therapy techniques. With the knowledge of heat effects on phages, control groups observe differences in bacteriophage effect, therefore eliminating immunologic response variation in experiments. The National Institute of Health has done extensive controls on phage therapy on Vancomycin-Resistant Enterococcus faecium in mice, where single injection of phage administered 45 min after bacterium contact rescued 100% of mice compared to 100% fatality when no phage was administered [m10]. Numerous studies have shown substantial efficacy of phage on gram negative bacteria in calves, pigs, mice, and lamb[soothill01].
 
:Extensive studies of phage therapy on gram negative bacteria have already been proved extremely effective [soothill, smith hw]. Experimentation of E. coli done on in vivo mice showed that bacteriophages were more efficient in vivo experimenation than in vitro. [59] Research on calves showed that one injection of specific phage strain 8 hours after contact with E coli protected the calves from death and diarrhoea, and again the in vitro experimentation underestimated the virulence of phage. Futhermore, different strains of phage were tested for efficiency in isolated does and cocktail formulas [60]. Some strains were proven more effective than leading antibiotics []. [soothill experiments needed].
 


== Questions and Specific Aims ==
== Questions and Specific Aims ==
:
[[Experiments]]
:
:'''Pilot Project Aims'''
:
:'''Specific Aims'''
<br />
<br />
1. Begin a long-term mobile screening program for antibiotic resistant gonorrhea on the streets of Oakland and San Francisco.<br />
:'''Isolate N. gonorrhoeae from 10 patients'''. 
:a. Develop a website through which patients, after being provided a patient number can retrieve their results with a full analysis of their strain.
:''Acquiring samples''. We are currently in negotiations with local clinics (ie. Berkeley Free Clinic, Magnet, City Clinic :of San Francisco and St. James Infirmary) to enroll patients in our pilot study. Briefly, patients who are screened by :these clinics and test positive for oropharyngeal N. gonorrhoeae will be provided with information on our study. Within :the informational packet, patients will find their assigned Patient ID and use this number to schedule an in-house visit. :The Patient ID allows the patient to remain anonymous during our screening process. During their primary visit, patients :will provide their consent, sign liability waivers, fill out our surveillance survey, and provide us with an :oropharyngeal specimen. Patients can then use their Patient ID to track the results from their specimen using our :anonymous web tracking system.
:b. Compose and deploy a mobile screening unit at various locations throughout Oakland and San Francisco.
 
:''Primary isolation of N. gonorrhoeae''. Primary gonococcal cultures will be grown on Modified Thayer-Martin (MTM) medium containing chocolate agar (5% SRBC), Vancomycin, Colistin, Nystatin and SXT. This medium allows selective growth of Neisseria species. Primary cultures will be propagated in tryptic soy broth (TSB) according to the standards set forth by the Centers for Disease Control (REF).  
 
:''Confirmation of N. gonorrhoeae''. Upon primary culture and propagation, N. gonorrhoeae samples will be confirmed using nucleic acid testing. A panel of polymerase chain reactions will be performed on DNA isolated from bacterial cultures. These panels will be performed to (1) identify genes responsible for 1st-, 2nd- and 3rd-generation antibiotic resistance, (2) differentiate N. gonorrhoeae from other Neisseria species. The differentiation PCR will be confirmed using CTA sugar growth testing as described by the CDC (REF). Briefly, isolates will be assessed for their ability to grow in Glucose, Maltose, Lactose and Sucrose. N. gonorrhoeae is positive for glucose metabolism, but not maltose or lactose metablism.
<br />
<br />
2. Support the Center for Disease Control's efforts to monitor the emergence of antibiotic resistant gonorrhea.
:'''Isolate ARNG-derived bacteriophage from 10 patients'''.  
:a. Compile data from the mobile screening program and report it (anonymously) to the CDC.
:''Bacteriophage Isolation''. Bacteriophage will be isolated from 10 clinical samples as previously described (REF). Briefly,  isolated colonies will be selected and cultured in  5 mL TPY broth (trypticase, phytone, and yeast extract) and cultued for 24 hours at 37oC in 5% CO2.
:b. Send antibiotic-resistant gonorrhea samples to the center for disease control according to the Instructions for Submitting Specimens to CDC Gonorrhea Laboratory for Confirmation Testing and/or Testing of Clinical Treatment Failures.
<br />
<br />
3. Develop a community laboratory space in Oakland, CA that has the equipment (acquired or built) we need in which we can conduct our research.
:After culture, 1500 µL of broth will be transferred to Trypticase Soy Agar (TSA) plates in triplicate (500uL each). One plate will be used for harvesting bacteriophage, a second plate will be used for propagating bacteriophage, and a third plate used for storing cultures. The remaining culture (3500uL) will be centrifuged for 10min at 2500 RPM. The supernatant will be removed and filtered through a 0.22 µm syringe filter. The syringe filters removes all cellular debris resulting in a pure bacteriophage culture. Bacteriopahge cultures will then be added to a bacterial lawns using 10-fold titrations to determine titers. Positive titers will result in plaque formation.
:a. Procure capital equipment and reagents needed for this project.  
:b. Procure reagents and equipment to begin molecular microbiological research.
:c. Open space up to provide education for Oakland teens.
<br />
<br />
4. Isolate, identify and characterize the first known bacteriophages from '''N. gonorrhoeae.'''
:'''Characterize ARNG-derived bacteriophage from 10 patients'''.
:a. Swab patients, grow samples on Modified Thayer Martin medium, broth cultures grown in Trypticase Soy Broth.
 
:b. Supernatants harvested and filtered through a 0.2um filter and analyzed by SDS-PAGE.
:''Genomic Molecular Weight Analysis''. Bacteriophage genomes will be extracted as previously described (REF). The genomes :will then be analyzed via gel-electrophoresis to determine approximate molecular weight(REF).
:c. Characterize phage via PCR
 
<br />
:''SDS-PAGE''. Purified bacteriophage lysates will be analyzed by denaturing SDS-PAGE as previously described. The SDS-:PAGE characterization assays will determine the molecular weight of proteins and assess their relative abundance. All :samples will be compared to positive control T4-phage panels.
5.Develop methods by which bacteriophages can be cultured and used as prophylaxis and therapy for gonoccocal infections.
 
:a. Test and optimize growth conditions for strains of antibiotic resistant gonorrhea from which bacteriophage can be isolated.
:''Host Range Studies''. Host range studies will be performed as previously described (REF). The purpose of this assay is :to determine the ability of ARNG-associated phage to infect different Neisseria species, bacterial from different genii :(I.e gram positive, and coliforms), and alternate clinical N. gonorrhoeae samples. Briefly, bacterial lawns of varying :host bacteria will be created and phage will be drop-tested and assessed for their ability to produce plaques in :different genii, species, or isolates cultures. We also propose a novel broth-culture assay in which OD readings are :taken before and after bacteriophage infection. Should our bacteriophage isolates maintain their capacity for lysis, OD :culture readings should drop significantly.
:b. Harvest phage and perform proof-of-concept, in vitro experiments in which gonorrhea is lysed with preparations of bacteriophage.
 
:'''Conduct proof-of-concept lysis experiments'''.
:''Infection Assay''. Following testing of each individual phage, a 10-sample phage cocktail will be generated and tested :for its ability to lyse clinical samples of N. gonorrhoeae. These methods are similar to "Host Range Studies," the only :difference being that instead of using single-phage samples, the proof-of-concept treatment will be a 10-phage cocktail :in which all bacteriophage samples are combined at (a) the same titer, and (b) in the same proportions. These assays are :previously described (REF).
 
== Anticipated Challenges to Our Approach ==
 
#rapid clearance of phages by human host and bacteria: bacterial strain resistance / anti-phage antibodies.
#patient safety: altering phage to prevent lysis of gram negative species.
#bacterium targeting and optimization: altering the binding kinetics of Phage adherence.
#narrow range of hosts
#achieving phage purity: while administering directly may be hazardous if bacteria is not removed.
#poor stability of phage: how do we preserve phage ex vivo and ensure that is viable upon administration?
#Addressing previous citizen science failures and conducting research with a greater adherence to quality.
#Preventing Lysogeny.


== Experiments and Anticipated Problems ==
===Protocol Documentation===
:We plan to begin acquiring clinical samples in April, 2013. Samples will be grown on Modified Thayer Martin agar supplemented with antibiotics. Colonies will '''(1)'''be stored in glycerol at -80oC, '''(2)'''grown in liquid culture for the assessment of bacteriophage production, '''(3)'''grown in Maltose to ensure we have cultured ''N. gonorrhoeae,'' and '''(4)'''grown in Fastidious Broth with 3rd-generation antibiotics to assess antibiotic resistance.
http://aegia.nu/folder/getitproject/130721_prephage-protocols.txt
:
:Following growth in FB, supernatants will be analyzed for the presence of bacteriophage using SDS-PAGE electrophoresis.
:
:If bacteriophage proteins are present in the supernatants, these cultures will be re-grown in bulk and bacteriophage-containing sups will be stored at -80oC.
:
:All clinical screening data will be available to patients online through our website. If patients are positive, they will be asked to fill out a form on their results page that will detail their risk level. This data is important in the surveillance of antibiotic-resistant ''N. gonorrhoeae'' emergence. '''''All screening procedures and results are anonymous.'''''


== Laboratory Needs ==
== Laboratory Needs ==
#CO2 incubator (we've built a prototype incubator but it has no CO2 component). We can grow these organisms in candle jars inside the incubator which means we need candle jars and candles.
:[https://www.wepay.com/donations/gonorrhea-eradication-team Your Financial Support]
#Benchtop butane torches.
:CO2 incubator (we've built a prototype incubator but it has no CO2 component). We can grow these organisms in candle jars inside the incubator which means we need candle jars and candles.
#Sterilized toothpicks.
:Benchtop butane torches.
#MTM agar plates '''(ordered!)'''
:Sterilized toothpicks.
#Fastidious Broth <ref>Cultivation of Neisseria gonorrhoeae in Liquid Media and Determination of Its In Vitro Susceptibilities to Quinolones. http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1234085/</ref>
:MTM agar plates '''(ordered!)'''
#Glass test-tubes.
:Fastidious Broth <ref>Cultivation of Neisseria gonorrhoeae in Liquid Media and Determination of Its In Vitro Susceptibilities to Quinolones. http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1234085/</ref>
#Maltose for differentiation of N. meningiditis vs. N. gonorrhoeae.
:Glass test-tubes.
#Rayon Swabs and tongue depressors ('''ordered!''')
:Maltose for differentiation of N. meningiditis vs. N. gonorrhoeae.
#-80c Freezer to store glycerol stocks of clinical samples.  
:Rayon Swabs and tongue depressors ('''ordered!''')
#Ultracentrifuge
:-80c Freezer to store glycerol stocks of clinical samples.  
#SDS-PAGE gel electrophoresis equipment and reagents.
:Ultracentrifuge
#Autoclave
:SDS-PAGE gel electrophoresis equipment and reagents.
:Autoclave
:
:
: '''''much much much more'''''.
: '''''much much much more'''''.
<br />
==Guiding Principles==


== Who We Are ==
:Throughout the course of this project, participants will co-create and foster an open access citizen science project that accomplishes specific scientific goals while educating and nurturing community and scientific creativity.
:Those participating in this project adhere to the following statements:
:  
:  
''We are building egalitarian networks.'':  
''We believe in building egalitarian networks'':  
: Hierarchy, privilege and discrimination have the potential to repress creative thought in science, deny access to science, and as a result, hold back global scientific development. As a collective of citizen scientists working toward a more complete understanding of molecular medicine, we commit ourselves to nurturing creative, positive voices within our community. We commit ourselves to listening to, hearing and acknowledging the diverse voices that make our community great!  
: Hierarchy, privilege and discrimination have the potential to repress creative thought in science, deny access to science, and as a result, hold back global scientific development. As a collective of citizen scientists working toward a more complete understanding of molecular medicine, we commit ourselves to nurturing creative, positive voices within our community. We commit ourselves to listening to, hearing and acknowledging the diverse voices that make our community great!  


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''We believe in asking (and answering) questions'':  
''We believe in asking (and answering) questions'':  
: '''Science is a ''quest''ion'''. Our social quest is the pursuit of knowledge.  We desire to collectively answer questions and serve our communities. We believe that all knowledge must be '''free''' and '''accessible''' and will continue to work to maintain that openness.
: '''Science is a ''quest''ion'''. Our social quest is the pursuit of knowledge.  We desire to collectively answer questions and serve our communities. We believe that all knowledge must be '''free''' and '''accessible''' and will continue to work to maintain that openness.
''We believe in equal access to healthcare.''


== Relevance ==
== Relevance ==
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== '''Meeting Announcements and Contact Information''' ==
== '''Meeting Announcements and Contact Information''' ==


'''''Next Call: 3/24/13 8:00pm PST/10:00pm CST'''''
'''''Next Call: 8/4/13 8:00pm PST/10:00pm CST'''''
<br />
<br />


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== References ==
== References ==
<references />
<references />
[[Category: Health]]
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