journal of continuing educationt iopics ssues · 2013. 4. 25. · figure 3. wide branching,...

Journal ofContinuingEducation

Official publication of the American Medical Technologists

April 2013Volume 15 ■ Number 2

TopiCs TopiCs issuEsissuEs

✒ Quality Control: Part 3

✒ Case Twenty Five: A Disseminated Fungal Infection

✒ The Role of Animal Models in Determining the Genetic Bases of Osteosarcoma

✒ Safely Storing Laboratory Chemicals

Inside This IssueInside This Issue

&&

For additional Information:Contact AMT: 10700 West Higgins Road, Suite 150Rosemont, IL 60018Phone 847/823-5169 • Fax: 847/823-0458E-mail: [email protected] Website: www.americanmedtech.org

Omni William Penn Hotel

AMT 75th Educational Program & National Meeting • July 8–12, 2013

PITTSBURGH, PENNSYLVANIA

AMT Convention room rate:

$ 119.00 plus 14% taxes (rate valid from

July 2 to July 15, 2013)

Omni William Penn Hotel530 William Penn Place, Pittsburgh, PA 15219Phone (412) 281-7100Website: http://www.omnihotels.com/

FindAHotel/PittsburghWilliamPenn.aspx

o know Pittsburgh, you have to see it for yourself. Come and see a city that has had a remarkable environmental renaissance, a top-10 city for certified green building space, a city ripe with natural and cultural amenities. • Pittsburgh International Airport is nation’s third largest airport.

• In a statement to the New York Times, Prince Phillip said that Pittsburgh is the only city he has ever seen that has an entrance. The first glimpse of the Golden Triangle upon exiting the Fort Pitt Tunnel, on the way in from the airport, is absolutely incredible!

• There are 50 museums located in and around the Pittsburgh area.• Pittsburgh, the “City of Champions,” is a sports fan’s paradise with a new football

stadium and new baseball park.• Located just a short drive from Pittsburgh is one of the most famous private

homes ever built, Fallingwater, Frank Lloyd Wright’s architectural masterpiece.

FEATURES:• Downtown location – many restaurants and shopping within walking distance• Located just 18 miles from airport• Self parking $13.75 weekdays, $5.00 weekends• Directly across the street from the “T,” Pittsburgh’s free underground rail system• Each guestroom has coffeemaker, hair dryer, iron and ironing board• When a guest joins the free Omni loyalty program, he/she will receive many

complimentary benefits during his/her stay, such as complimentary Wi-Fi service, morning beverage delivery, pressing of clothing items, bottled water, shoe shine, newspaper.• AMT attendees will receive 15% discount in all hotel restaurants.

Contents34 AMTIE President’s Report

36 Cases in Clinical Microbiology

38 Article 398 Quality Control: Part 3

David Plaut and Deena Davis

41 Questions for Article 398

42 Crossword Puzzle

44 Article 399 Case Twenty Five: A Disseminated Fungal Infection

Irmeen Siddiqui, Carlos Teran, Katie Ruger and Joel Mortensen


50 Article 400 The Role of Animal Models in Determining the Genetic Bases

of Osteosarcoma Haider Saddam Qasim

Center Pullout SeCtion

Pittsburgh Convention Preliminary Program


66 Article 401 Safely Storing Laboratory Chemicals

Karen Appold


70 AMT Directory

72 Abstracts From the Current Literature

Journal of Continuing Education Topics & Issues (ISSN 1522-8606) is published in January, April, and August under the sponsorship of the American Medical Technologists, 10700 W. Higgins Rd., Suite 150, Rosemont, Illinois 60018. Copyright 2013 by American Medical Technologists. Subscriptions include three issues of Journal of CE Topics & Issues and four issues of AMT Events: $50.00/year + $10 postage for foreign countries. Members may not deduct subscription price from dues.Postmaster: Please send change of address to AMT, 10700 W. Higgins Rd., Suite 150, Rosemont, Illinois 60018.Moving? Be sure AMT publications move with you. Send your new address and old mailing label from an AMT publication to AMT six weeks before you move.

Cover photo: Hydrocortisone photomicrograph, © Eric Clark, National High Magnetic Field Laboratory, Florida State University, Tallahassee.

EditorGerard P. Boe, PhD

Associate EditorDiane Powell

Business OfficeAmerican Medical Technologists10700 W. Higgins Rd., Suite 150Rosemont, IL 60018847-823-5169e-mail address: [email protected] Site: http://www.americanmedtech.org

Journal ofContinuingEducationApril 2013Volume 15 ■ Number 2

TopiCs issuEs&

34 April 2013 • Continuing Education Topics & Issues

Amer ican Medical Technologists Inst i tute for Excel lence (AMTIE)

PRESIDENT’S REPORTAMTIE’s New Direction

Recently I received a copy of AMT's new Annual Report, entitled “American Medical Technologists October 2011- September 2012, A Year of Progress.” It is an exciting overview of the growth and im-provements of the American Medical Technologists organization and included information about the American Medical Technologists Insti-tute for Excellence (AMTIE). AMT has grown in membership not only in the US but also to include members in 56 countries. Because of this growth and the need to provide educational opportunities to its mem-bers, the role of AMTIE has changed.

AMTIE is a 501(3)(c) [tax exempt] educational support corporation charged with the ad-ministration of scholarships and grants associated with enhancing educational and competency opportunities for current and future allied health professionals. AMTIE solicits contributions from its members and is in the process of getting its strategic initiatives underway to solicit donations from businesses, philanthropic organizations, and non-AMT members who have expressed the desire to support our worthy cause. In 2012, AMTIE gave seven scholarships, totaling $6,500, and awarded $1,000 in educational grants/awards.

AMTIE Board of Trustees Election

Two open positions are available in 2013. The June issue of AMT EVENTS will publish candidates' notice of intent and photos. This information must be submitted to Diane Powell at the AMT Home Office immediately. The final deadline to submit your application to the AMT office staff and have it posted on the bulletin board in the AMT Registration area of the conven-tion is Monday, July 8th, by 10am. NO EXCEPTIONS. The voting will take place on Wednes-day, July 10th, and all AMT members can vote for the two candidates of their choice. You do not have to be a delegate to vote in the AMTIE election. All applicants MUST present them-selves to the membership at "Meet the Candidates" on Tuesday, July 9th.

AMT Presents the 75th Educational and National Meeting

The Omni William Penn Hotel, Pittsburgh, PA, is the location for the national convention. The dates are July 8-12, 2013.

• This year the meeting will be a day shorter, but nonetheless filled with educational oppor-tunities for your continuing education to meet recertification requirements.

• The Welcome Reception will be held Monday night with music, dancing, and a lot of fun.

• The Awards Banquet will be on Wednesday night. At this time, we honor outstanding AMT members.

Linda Jones, MT, AMTIE President

Continuing Education Topics & Issues • April 2013 35

• The week is filled with committee meetings, town hall, district meetings, the seating of the delegates with voting for bylaws changes and national board members.

• It is a jam-packed meeting to include the AMT family reunion. There will be lots of hugs from old friends and the opportunity to make new acquaintances.

• … and lastly, but most important — AMTIE is sponsoring the “Boasting Arts and Crafts Show.” We are asking AMT members to donate one of their crafts to be sold at auction. The money from the sale of the items will be donated to AMTIE to help sponsor the scholar-ships and grants awarded by AMTIE. Taffy Durfee, our AMTIE Trustee from Texas, is heading up the sale. Please let her know if you plan to contribute to the Arts and Crafts Show. [email protected] or call 979-776-2571.

Thank you for allowing me to serve you as I am most proud of our organization and profession.

Best regards, Linda Jones, BS, MT


Case Description: An11-year-old male, who previously had a bone marrow transplant (April 2012) secondary to X linked hyper-IgM syndrome, developed skin Graft-versus-Host Disease shortly

after bone marrow transplant and was placed on azithromycin and acyclovir. Due to persistent gastrointestinal symptoms including hematemesis (blood in his vomit), he was readmitted in June 2012. Because of the immunosuppression, history of histoplasmosis lymphadenitis, severe giardiasis and CMV viremia in the past, he was placed on broad-spectrum antibacterial agents (ciprofloxacin, meropenem, metronidazole, and vancomycin), antifungal agents (caspofungin and voriconazole) along with a prophylactic antiviral agent (gancyclovir). During this stay, he started developing respiratory distress and fever. Multiple blood cultures were performed, but all remained negative after 5 days. CT scan of the chest showed the presence of multiple pulmonary nodules and effusion. Bronchoalveolar lavage specimens demonstrating lipid and hemosiderin laden macrophages and respiratory cultures showed few epithelial cells and white blood cells. No organisms were seen. Fungal work-up was also negative including urine antigen testing for Histoplasma Antigen performed by quantitative sandwich enzyme immunoassay at Mira Vista Diagnostics, Indiana, and serum Cryptococcal antigen performed using semiquantitative enzyme immunoassay at ARUP laboratories, Utah.

Endoscopic studies (August 2012) were performed because of continuing gastrointestinal symptoms including diarrhea, nausea, vomiting showed the presence of fungal hyphae on periodic acid-Schiff stained sections of gastric mucosa. The sample was not cultured at that time.

Cases in Clinical Microbiology

The case description on this page and its follow-up discussion presented elsewhere in this issue is the 25th in a series of articles presenting clinical microbiology cases that will appear in this journal. Readers should study the case de-scription below and formulate their own answers to the questions posed. After coming up with a solution to the problem, turn to page 44 in this issue and read the Case Fol-low-up and Discussion. This is followed by Questions for STEP Participants on page 48.

Joel E. Mortensen, PhD, MLT(AMT), Series Editor

Figure 1(a).Gastric mucosa (thick arrow) with fungal hyphae (thin arrow) (X20 Periodic acid-Schiff stain)

Figure 1(b). Broad, ribbon-like aseptate fungal hyphae (X40 Periodic acid-Schiff stain)


Hematoxylin and Eosin stained section of the tissue from the maxillary sinus showed abundant invasive fungi with broad aseptate hyphae.

In the laboratory, maxillary sinus contents were inoculated onto two Sabouraud dextrose with Brain Heart Infusion agar plates (SABHI) for the isolation of fungi. On day five, a wooly-dark mould grew on both SABHI plates. Lactophenol blue tape preparations revealed globose sporangia with rhizoids directly under unbranched short sporangiophore.

The patient was placed on amphotericin B liposomal but was persistently febrile and developed seizures. As part of the search for a focus, a head CT scan was performed, showing the presence of acute sinusitis of the sphenoid, ethmoid and maxillary sinuses with new subperiosteal abscess inferior medial left orbit. CNS infiltration was also considered due to the seizures. The patient was taken to the operating room every few days for surveillance endoscopy and debridement of the advancing sinusitis.

What is the most likely identification of this mold?

Figure 2(a): Head CT scan showing sub periosteal abscess in the inferior medial left orbit (yellow arrow) and sphenoidal sinusitis (red arrow).

Figure 2(b): Brain MRI showing the exenterated left eye without intracra-nial extension.

Figure 3. Wide branching, aseptate fungal hyphae (X40 hematoxylin and eosin stain)

Figure 4(a). Fungal organism on Sab-ouraud dextrose with brain heart infu-sion agar at day 5 and 30°C.

Figure 4(b). Globose sporangia (long arrow) with rhizoids (thick arrow) di-rectly under unbranched sporangio-phores (short arrow). (Katie Ruger, 2012).

See page 44 for case followup and discussion.


Welcome to the third installment of our series on Quality Control. In this installment we will discuss two sets of QC rules that will allow you to monitor and respond quickly to legitimate out of control situations as opposed to the false re-jects that are so frustrating, not to mention time consuming and expensive.

In this session we will take the mean and SD that we discussed in the last installment and use them with a QC chart (also called a Levey- Jennings or L-J chart). We will illustrate two ways to plot the two controls: 1) using one piece of paper or one worksheet in Excel and 2) using a single sheet for both controls. We leave it to you to choose the one you prefer. Our choice is the single sheet as it is easier to see both con-trols concurrently. Figure 1 is a chart for a single control over 30 runs. There is nothing magic about the 30 runs, although most interlaboratory programs report data each month (30 days in a month with one run of QC per day equals 30 runs total). The chart for the second level would be similar. Figure 2 shows both controls on one sheet. We have omitted the SD lines for clar-ity. You would want them on yours. We have plotted the mean and the + and – 2, and 3 SD lim-its on the chart in Fig. 1. We use these limits to aid us in determining whether to label a run “in” or “out.” These are indeed labels as we are never 100% certain whether our label is correct. You may recall in school sometimes hoping the teacher would “grade on the curve.” It is this curve (often called a bell or normal curve) we use to label a run. Figure 3 is a drawing of the curve.

Note the shape of the curve. Although it ap-pears that the curve touches the x-axis (the hori-zontal line), it only comes extremely close to it, but never really touches it. Most of the area (68%) lies within the ±1 SD from the mean (0 in Figure 3), 95% of the area lies within ±2 SD and

99.7% lies within ± 3 SD of the mean. This is true of any of the measures we make repeatedly on the control material (or a patient sample) while the system is stable. Stable in the sense that the instrument is performing as it should and no changes are made in reagent or calibration during a given period. There will always be some variation (measured by the SD) even when the instrument is performing well. This is why there are limits rather than expecting all the data to be on the mean. (You may want to plot a histogram of data from one or more of your controls — say on glucose or hemoglobin — to see how closely your data fit the curve. It works best to use a dot plot with at least 70 points.)

We use the probabilities from the curve to de-termine whether to label a run “in” or “out,” knowing that it is a probability and not a certain-ty. For example when we label a value that is more than 3 SD from the mean "out," we are say-ing we are more than 99% sure that there is a problem somewhere in the system. It could be the control or the reagents or the calibration or the instrument. Until we stop to ponder why the value(s) exceeded our limits, we don't know what is needed to fix the problem.

We use these probabilities with the two or three controls to make our decisions about the run. We will illustrate each of the rules we rec-ommend and discuss in some detail what a rule

David Plaut, Plano, Texas, Consultant, AMT’s Book Re-viewer, and frequent speaker at AMT na-tional conventions and regional meet-ings; Deena Davis, MLS, Point of Care Coordinator for Bozeman Deacon-ess Hospital in Boz-eman, Montana.

Quality Control: Part 3

David Plaut and Deena Davis

Article 398 1 Clock Hour

Figure 1


failure can tell us about the source of the problem, in other words, knowing which rule was broken (the run labeled out). In our next installment, we discuss a bit more about trouble shooting when we have data from an instrument that measures more than one constituent at the "same" time.

The flow chart in Figure 4 is an easy way to im-plement the rules we recommend. These rules are a combination of rules suggested in 1974 by the CAP and in 1981 by Westgard et al. It is worth not-ing that these rules have been validated to detect virtually all of the errors that would cause a profi-ciency failure or lead to a clinician making a wrong

decision (based on laboratory data). If, and only if, you are doing your QC manual-

ly — charting the controls on paper by hand — the first rule or flag to consider is the 1 2SD flag. If there is no control value greater than 2 SDs from the mean, the run is labeled 'in' and the patient data are accepted. If there is a control value outside the 2 SD limits (a value on the 2 SD line is NOT out.), you need to look further. As the flow chart indicates, the next question is whether there is a control outside 3 SD. Figure 5 shows an example of a single control outside 3 SD. If so, the run is put on hold, all patient results should be held and trouble shooting is begun. Trouble shoot-ing does NOT begin by rerunning the control. Re-running the control when one of these rules is bro-ken is simply a waste of money and time. A violation of a rule in the flow chart is not an error that will simply go away, no matter how hard you wish for it to or how many times you rerun the control. Some good old fashioned problem solving is the only answer. If neither control is outside 3 SD, continue along the flow chart to see if a sec-ond control is outside 2 SD on the same side of the mean. Figure 6 illustrates this 2 2 SD failure within a run. If a 2 2SD failure occurs, stop and begin trouble shooting.

Figure 4

Figure 5 Figure 6

Figure 2 Figure 3


If the second control is not outside the same (+ or –) 2 SD limit as the first control, then look at the previous run to see if the control that is out on this run was out on the previous run. If that is the case, stop and trouble shoot. Figure 7 illustrates this 2 2 SD failure across runs. If there are not con-secutive values outside the same 2 SD limits (2 se-quential results out by 2SD on the same side of the mean), on the current run where one control is outside 2 SD, look to see if the other control is also outside the other 2 SD limit. Figure 8 illustrates this. If this rule, the R 4SD rule, is not violated, all that has happened is one control is outside 2 SD which is NOT cause to stop. Report the data, no troubleshooting is necessary.

As we mentioned, we will discuss trouble shooting in the next installment and we will also discuss ways to deal with repeating certain pa-tient samples.

If a run is rejected as ‘out,’ do not include the control data in the ‘good’ file. Do not discard it completely, but do not include it with the data used to calculate the mean and SD. Simply keep it so you may refer back to it in case this error oc-curs again and/or becomes a persistent problem.

If you use three controls, the only rule that changes is the 2 2 SD within a run which becomes the 2 of 3 2 SD rule. This is illustrated in Figure 9.

The reasons we recommend these rules, aside from the fact that they have been recommended by others, is that they are very sensitive to signifi-cant errors, those errors that could lead to harm to

the patient or proficiency failures. They have about one-tenth of the false alarms that the 1 2 SD error has (rejecting a run with a single value out-side 2 SD). They are simple to use manually and they can aid in trouble shooting. We can have greater faith in our analyzers and thus provide our patients with safe and accurate lab results.

References1. CAP Today, 19742. J O Westgard and T Groth Design and evaluation of statistical

control procedures: applications of a computer "quality control simulator" program.Clinical Chemistry 1981; v. 27, p.1536-45.

3. D D Koch, J J Oryall, E F Quam, et al. Selection of medically useful quality-control procedures for individual tests done in a multitest analytical system.Clinical Chemistry 1990; v. 36, p.230-3.

4. J O Westgard, P L Barry, M R Hunt, et al. A multi-rule Shewhart chart for quality control in clinical chemistry.Clinical Chemistry 1981; v. 27, p.493-501.

Figure 7 Figure 8

Figure 9


1. QC rules exist to ________ a run. A. Assign B. Label C. Identify D. Allocate

2. A QC chart uses these statistics: A. Mean and CV B. CV and SD C. Mean and SD D. Mean, SD and CV

3. Runs are called “in” or “out” based on the QC.

A. True B False

4. If a run is out, first rerun the control that was out.

A. True B False

5. When the system in working as best it can, values will be outside 2 SD.

A. 0.3% B. 5.0% C. 16% D. 95%

6. The QC rules suggested in this article date from

A. Landsteiner – early 1920s B. Levey and Jennings – mid 1950s C. CAP and Westgard – 1970-80 D. CLIA – 1990

7. The ‘normal’ or bell or Gaussian curve illustrates

A. Systematic shifts B. Random error C. Both a and b D. Neither a nor b

8. Two values outside the same 2SD limit points to a

A. Random error B. Systematic error C. Either random or systematic error D. No error

9. A single value outside 3 SD points to a A. Random error B. Systematic error C. Either random or systematic error D. No error

10. If QC is done manually, controls outside 2 SD should be rerun.

A. True B False

Questions for STEP ParticipantsAMT strongly encourages you to submit your answers online so that the CE credits can be auto-

matically transferred into your AMTrax account. To do so, go to www.americanmedtech.org, click on the AMT Store on the top navigation bar. Click on STEP Online, then select the article number and purchase the test. Don’t forget to log in to receive the discounted member price of $5 (nonmembers pay $15/test).

If you wish to submit answers manually (only available to AMT members), the fee is $10/test. Please submit a copy of this page with your answers marked, along with a completed order form lo-cated elsewhere in this publication. Don’t forget to include payment.



Crossword Puzzle

Answers on page 68


Case Follow upThe organism was presumptively identified as

Rhizopus species based on the morphology of sporangiophore and sporangia. The Fungus Testing Laboratory, University of Texas Health Science Center later confirmed the identifica-tion as Rhizopus microsporus using growth characteristics and DNA sequencing. Of serval varieties of Rhizopus microsporus, the closest variety that resembled was R. microsporus vari-ety chinensis.

The subperiosteal abscess spread into the orbit and eye tissue and subsequent follow up MRIs and CT showed central nervous system spread of the lesions, as well. The Ophthalmolo-gy service performed a exenteration of the in-volved left eye because of the advancing infiltra-tion. Because of the central nervous system involvement and mental status deterioration, he was intubated and placed on cardiorespiratory support. His clinical status continued to deterio-rate, requiring more respiratory support with higher setting of the ventilator and finally died almost two months after admission.

BackgroundZygomycosis infections have been reported

worldwide and are the third most common inva-sive fungal infection after candidiasis and asper-gillosis3. Rhizopus, together with Mucor, Rhizomucor, Absidia, Apophysomyces,

Cunning hamella, Syncephalastrum and Sakse-naea are placed in the order of Mucorales, class of Zygomycetes1. The incidence of zygomycosis is rising due to multiple factors including the he-matological malignancies, immunosuppression, stem cell and organ transplants1.

Mucorales are ubiquitous moulds, widely dis-tributed in the environment including soil, plants, decaying organic matter4. They are fre-quent contaminants of food and grow well on fruits, cereal and bread3. Airborne spores are thought to be the infectious particles responsi-ble for disease, particularly in immunocompra-mised individuals and this explains the frequent localization in skin, sinuses and lungs. These molds are often found as laboratory contami-nants and may cause nosocomial infection5.

The genus Rhizopus is characterized by rapid production of white fluffy colonies which turn brownish to black with time due to the presence of pigmented sporangiophores and sporangia5. Rhizopus microsporus and Rhizopus oryzae are the most common species involved in zygo-mycosis5. Rhizopus microsporus is a thermo-philic zygomycete and can be isolated from soil or wood products. Different varieties of Rhizo-pus microsporus include Rhizopus microspo-rus var microsporus, Rhizopus microsporus var chinensis, Rhizopus microsporus var oli-gosporus and Rhizopus microsporus var rhizo-podiformis. These can be differentiated on the basis of their sporangiospores morphology and the growth at different temperatures5. Among these known four varieties, only Rhizopus mi-crosporus var microsporus and Rhizopus mi-crosporus var have been reported to cause in-fections in humans3.


Dr. Siddiqui is a Pathology Resident in the Department of Pathology and Laboratory Medicine of the University of Cincinnati. Dr. Teran is a Fellow in the Division of Infectious Diseases at Cincinnati Children’s Hospital. Katie Ruger is Medical Technologist in the Diagnostic Infectious Diseases Testing Laboratory in the Department of Pathology and Laboratory Medicine at Cincinnati Children’s Hospital. Dr. Mortensen is the Director of Diagnostic Infectious Diseases Testing Laboratory in the Department of Pathology and Laboratory Medicine at Cincinnati Children’s Hospital, and Professor in the Department of Pathology and Laboratory Medicine of the University of Cincinnati, Ohio.

EDITOR’S NOTE: BEFORE reading the Case Follow-up and Discussion below, study the Case Description on page 36 of this issue, and formu-late your own answers to the questions posed.

Case Twenty Five: A Disseminated Fungal InfectionIrmeen Siddiqui, Carlos Teran, Katie Ruger and Joel Mortensen

C A S E S I N C L I N I C A L M I C R O B I O L O G Y


Clinical PresentationLocalized and disseminated forms of the dis-

ease need to be differentiated as the two most common initial presentations are rhinocerebral or pulmonary infection which rapidly progresses and involve the other viscera8. Other common forms recognized are orbital, cutaneous, gastro-intestinal and disseminated3. Disseminated rates vary from 3% to more than 50% for patients with hematological malignancies (depending to some extent on the underlying diseases and the site of infection)7, 8, 9. Sinus involvement is the most common presentation in the patients with diabe-tes and intravenous drug abusers and the pulmo-nary infection is the second most common while the reverse is true for hematology patients7, 10. In-fection causes necrosis and hemorrhage, local-ized or disseminated, which can have both clini-cal and radiological similarity to invasive aspergillosis16.

Cerebral infection may complicate sinusitis or can be the only presentation, especially in IV drug abusers; usually the patients present with lethargy, confusion or focal neurological deficits.

Gastrointestinal involvement has been de-scribed in 7% of cases especially in low birth weight premature infants or malnourished indi-viduals which can present with abdominal pain, perforation, peritonitis and dissemination to the liver7.

PathogenesisThere are several factors responsible for the

pathogenicity of different fungi. Macrophages and neutrophils play an important role in pro-tecting against zygomycosis. Prolonged neutro-penia or impaired function of neutrophils and macrophages induced by any drug (corticoste-roids) or underlying disease like diabetes are important risk factors for zygomycosis18. Studies have also pointed an important role for iron in these infections. Iron is essential for the viru-lence and growth of Mucorales so the conditions associated with iron overload like hematologic malignancies or hemodialysis with deferroxam-ine therapy can be predisposing factors for the infection19. Adhesion to endothelial cells and in-ternalization of fungus facilitates the process of angioinvasion causing infarction of surrounding tissues and dissemination into other organs20. Recently, a study has been done which demon-strated a novel host receptor (the glucose-regu-

lated protein 78 [GPR78]), which facilitates the invasion of human endothelial cells, by Rhizo-pus oryzae25. They found that in patients with iron overload and increase blood glucose (diabe-tes), there is increased expression of GPR78 re-ceptor and blood vessel destruction which is also the mechanism to gain access to central nervous system from direct extension and ves-sel invasion from paranasal sinuses25

Lab identificationDirect examination and histopathology:

Demonstration of hyphae in clinical samples is important for the diagnosis of zygomycosis11. Wet mounts of clinical samples can be observed directly by adding a few drops of potassium hy-droxide. Tissue samples can also be processed in the histopathology laboratory and hyphae can be visualized by staining sections by Gomori-meth-enamine-silver (GMS) or periodic acid-Schiff (PAS) staining. The specific morphological char-acteristics suggestive of Mucorales are thin, hya-line, non or pauciseptate walls, ribbon like hy-phae with large diameter and wide branching angles (>=90°)5. Culture is of prime importance for the diagnosis of zygomycosis because differ-entiating it from other mucorales and hyaline hyphomycetes can be difficult based on the mor-phology of hyphae in the tissue alone and also culture is needed for antibiotic susceptibility testing7. Mucorales are not fastidious organisms and can grow on common microbiological agar media, but mycological media like Sabouraud dextrose agar containing antibiotics are pre-ferred because it inhibits the bacterial growth and enables the isolation of some opportunistic fungi which are sensitive to cycloheximide, in-cluded is some selective fungal media5. Culture should be incubated at 37°C for optimal yields but for incubation of a second tube at 25° to 30°C is recommended because some species have an optimal temperature of growth below 37°C6.

Historically, identification of Mucorales fungi to the species level is based mainly on the exam-ination of their macroscopic and asexual micro-scopic characteristics or maximum growth tem-perature6. Genus Rhizopus produces pale to brownish cottony colonies. The sporangio-phores are unbranched; arise singly or in groups with well-developed rhizoids at the base. Rhizo-pus microsporus have sporangiophores arising from stolons and measuring up to 400 microme-ter in length. Sporangia are dark brown, colu-


mellae are conical and sporangiospores are stri-ate and angular to ellipsoidal. Rhizopus microsporus var chinensis have angular spor-angiospores with growth occurring at tempera-ture from 30° to 45°C5, 17.

There are currently no specific antigen detec-tion methods and there are no commercially available molecular detection techniques13. Dif-ferent approaches have been tried. Most studies have used PCR based techniques. A real time PCR assay targeting a conserved region of the Mucorales cytochrome b gene had a sensitivity and specificity of 56 and 100% respectively14. An in situ hybridization technique using Mucora-les-specific DNA probes targeting 18S rRNA sub-unit also showed promising results15. Chromo-somal sequencing of selected genes is becoming more common and probably represents the fu-ture of mould identification. In addition, the use of mass spectral analysis is being applied to the identification of moulds including zygomcetes.

Antifungal and antifungal susceptibility testing

Antifungal susceptibility can be evaluated both invitrio and in vivo. In vitro antifungal susceptibility can be evaluated by microdilution broth technique developed by Clinical and Labo-ratory Standards Institute21 or European Com-mittee on antimicrobial susceptibility testing22 or by commercially available E test23. Amphotericin B is the most active drug against Mucorales. Among the azoles, voriconazole has poor activi-ty whereas posaconazole has better activity with low minimum inhibitory concentration values5. Capsofungin has also shown efficacy in treating zygomycosis infection specially if combined with amphotericin B24.

ConclusionZygomycosis represents a fulminant fungal in-

fection especially in the immunocompramised patients or with diabetes. Rhizopus microspo-rus and Rhizopus oryzae are the most common causative agents of zygomycosis. The disease typically involves the rhino-orbital-cerebral area, lungs, gastrointestinal system, skin and less commonly other organs too. Timely diagno-sis of zygomycosis by culture and histology fol-lowed by surgery and correction of underlying condition is the key for successful treatment. Posaconazole and amphotericin B are usually preferred as a first line of therapy.

References1. Uckay I, Chalandon Y, Sartoretti P, Rohner P, Berney T, Ha-

daya K and van Delden C. Invasive zygomycosis in transplant recipients. Clin Transplant 2007: 21: 577-582.

2. Cheng VCC, Chan JFW, Ngan AHY, To KKW, Leung SY, Tsoi HW, Yam WC, Tai WM, Wong SSY, Tse H, Li IWS, Lau SKP, Woo PCY, Leung AYH, Lie AKW, Liang RHS, Que TL, and Yuen KY. Outbreak of Intestinal infection due to rhizopus microsporus. Journal of Clinical Microbiology. 2009:47: 2834-2843.

3. De Mol P and Meis JFGM. Disseminated Rhizopus microspo-rus infection in a patient on oral corticosteroid treatment: a case report. The Journal of Medicine. 2009:67:25-28.

4. Ribes JA, Vanover-Sams and Baker DJ. Zygomycetes in human disease. Clin. Microbiol. 2000: 13: 236-301.

5. Gracia-Hermoso D, Dannaoui E, Lortholary O and Dromer F. Agent of systemic and subcutaneous mucormycosis and ento-mophthoromycosis. Manual of Clinical Microbiology. 20--Chapter 119: 1880-1901

6. Schwartz P, Lortholary O, Dromer F and Dannui E.Carbon assimilation profiles as a tool for identification of zygomy-cetes. J. Clin. Microbiol. 2007: 45: 1433-1439.

7. Roden MM, Zaoutis TE, Buchanan WL, Knudsen TA, Sark-isova TA, Schaufele RL, Sein M, Sein T, Chiou CC, Chu JH, Kontoyiannis DP and Walsh TJ. Epidemiology and outcome of zygomycosis: a review of 929 reported cases. Clin. Infect. Dis. 2005:41: 634-653.

8. Nosari A, Oreste P, Montillo M, Carrafiello G, Draisci M, Muti G, Molteni A and Morra E. Mucormycosis in hematolog-ic malignancies: an emerging fungal infection. Hematologica. 2005:85: 1068-1071.

9. Pagano L, Offidani M, Fianchi L, Nosari A, Candoni A, Picar-di M, Corvatta L, D'Antonio D, Girmenia C, Martino P and De Favero A. Mucormycosis in hematologic patients. Hematolog-ica. 2004: 89: 2-7-214.

10. Kontoyiannis DP, Lionakis MS, Lewis RE, Chamilos G, Healy M, Perego C, Safdar A, Kantarjian H, Champlin R, Walsh TJ, and Raad II. Zygomycosis in a tertiary care cancer center in the era of Aspergillus-active antifungal therapy: a case control observational study of 27 recent cases. J. Infect. Dis. 2005:191: 1350-1360.

11. Torres-Narbona M, Guinea J, Martinez-Alarcon J, Munoz P, Pelaez T and Bouza E. Workload and clinical significance of the isolation of zygomycetes in a tertiary hospital. Med. Mycol. 2008: 46: 225-230.

12. Ruchel R and Schaffrinski M. Versatile fluorescent staining of fungi in clinical specimens by using the optical brightener Blankophor. J. Clin. Microbiol. 1999: 37: 2694-2696.

13. Dannoui E. Molecular tools for identification of zygomycetes and diagnosis of zygomycosis. Clin. Microbiol. Infect. 2009:15 (suppl.5):66-70.

14. Hata D, Buckwalter SP, Pritt BS, Roberts GD and Wengenack NL. Real-time PCR method for detection of Zygomycetes. J. Clin. Microbiol.2008:46:2353-2358.

15. Hayden RT, Qian X, Procop GW, Roberts GD and Lloyd RV. Insitu hybridization for the identification of filamentous fungi in the tissue section. Diagn. Mol. Pathol. 2002:11:119-126.

16. Sun HY, Aguado JM, Bonnati H, Forrest G, Gupta KL, Safdar N, John GT, Pursell KJ, Munor P, Patel R, Fortun J, Mar-tin-Davila P,Philippe B, Philit F, Tabah A, Terzi N, Chatelet V, Kusne S, Clark N, Blumberg E, Julia MB, Humar A, Houston C, Lass-Florl C, Johnson L, Dubberke ER, Barron MA, Lortholary O and Singh Pulmonary zygomycosis in solid organ transplant recipients in the current era. Am. J. Trans-plant. 2009:9:2166-2171.

17. Schipper MAA and Stalpers JA. A revision of genus Rhizopus.II. The Rhizopus microsporus group. Stud. Mycol. 1984: 25:30-34.


18. Morace G and Borghi Invasive Mold Infections: Virulence and Pathogenesis of Mucorales.International journal of microbiol-ogy. 2012

19. Symeonidis AS. The role of iron and iron chelators in zygo-mycosis. Clinical Microbiology and Infection. 2009: 19: 26-32.

20. Ibrahim AS, Spellberg B, Avanessian V, Fu Y, and Edwards JE. Rhizopus oryzae adheres to, is phagocytosed by, and damages endothelial cells in vitro. Infection and Immunity. 2005:73: 778-783.

21. Clinical and Laboratory Standards Institute (CLSI).Reference method for broth dilution antifungal susceptibility testing of filamentous fungi: Approved standard. Document M-38A. CLSI, Wayne PA

22. Rodriguez-Tudela JL, Donelly JP, Arendrup MC, Arikan S, Barchiesi F, Bille J, Chryssanthou E, Cuenca-Estrella M, Dan-noui E, Denning D, Fegeler W, Gaustad P, Lass-Florl C, Moore C, Richardson M, Schmalreck A, Velegraki A and Ver-weij PE.EUCAST technical note on the method for the deter-mination of broth dilution minimum inhibitory concentrations of antifungal agents for conidia forming moulds. Clin. Micobi-ol. Infect. 2008:14: 982-984.

23. Espinel-Ingroff A. Comparison of three commercial assays and a modified disk diffusion assay with two broth microdilu-tion reference assays for testing zygomycetes, Aspergillus spp., Candida spp., and Cryptococcus neoformans with po-saconazole and amphotericin B. J. Clin. Microbiol. 2006:44:3616-3522.

24. Reed C, Bryant R, Ibrahim AS, Edwards Jr. J, Filler SG, Gold-berg R, and Spellberg B. Combination polyene-capsofungin treatment of rhino-orbital-cerebral mucormycosis. Clin. In-fect. Dis.2008: 47:364-371.

25. Liu M, Spellberg B, Phan QT et al. The endothelial cell recep-tor GRP78 is required for mucormycosis pathogenesis in dia-betic mice. Journal of Clinical Investigation. 2010: 120: 1914-1924.

For more information please visit: www.americanmedtech.org

AMTrax ™

AMTrax is a member service designed to help you keep track of earned continuing education (CE) credit. The system:

Provides an effective means ofvalidating and organizing participationin CE activities

Provides a convenient means fordocumenting certification-relatedactivities for employers and licensingagencies

Helps self-assess progress incomplying with the CertificationContinuation Program (CCP)

Entering your CE data online is simple!Visit www.americanmedtech.org and clickon AMTrax under the CE tab. You must belogged in as a member to take advantageof this service.

AMTrax™

Online Recording System


1. Which of the following genera do not belong to the order Mucorales?

A. Mucor B. Rhizomucor C. Absidia D. Aspergillus

2. Which of the following clinical conditions represents the most common clinical presentation of zygomycosis?

A. Pulmonary and gastrointestinal B. Rhinocerebral and pulmonary C. Rhinocerebral and skin D. Disseminated

3. Which of the following mechanisms is not attributed to the pathogenesis of zygomycosis?

A. Impaired function of neutrophils and macrophages

B. Iron overload C. Lymphatic permeation and spread D. Angioinvasion and destruction

4. Which is considered as the most effective antifungal drug against Mucorales?

A. Amphotericin B B. Voriconazole C. Fluconazole D. Nystatin

5. Which of the following is a characteristic of Rhizopus species?

A. Narrow, septate hyphae with right angle branching

B. Broad based budding yeast C. Budding yeast with a halo around it. D. Broad, aseptate hyphae with wide angle

branching

6. The temperature of incubation is important for the isolation and identification of Zygomyctes. Media should be incubated at 37°C. and a second set at which of the following temperatures?

A. 35°C. B. 42 - 45°C. C. 25 - 30 °C. D. 5 - 10°C.

7. Which of the following represents the most common mode of transmission of Rhizopus infection?

A. Feco-oral B. Airborne C. Sexually transmitted D. Water borne

8. Which of the following statements best describe Rhizopus microsporus and Rhizopus oryzae?

A. the most common species involved in zygomycosis

B. always susceptible to amphotericin B C. cause diseases in children D. All of the above

9. Which of the following terms is not associated with Rhizopus microsporus?

A. Oligosporus B. Chinensis C. Fumigatus D. Rhizopodiformis

10. When stained with hematoxylin and eosin, which of the following statements best describes a Zygomycetes in tissue?

A. Narrow, septate hyphae with right angle branching

B. Broad based budding yeast C. Budding yeast with a halo around it. D. Broad, aseptate hyphae with wide angle

branching





STEP Article Number

Title Member Cost($10 per Article)

To submit STEP answers manually (only available to AMT members), you must complete this form and attach the appropriate quiz(es) with your payment and mail to American Medical Technologists.

The fee for manual submission is $10 per quiz. AMT will record your results in your AMTrax™ account.

NOTE: Florida state licensed MT or MLT members, if you would like AMT to record STEP article to CE broker please fill in your Florida state license number below.

STEP Order Form

Payment Information:

Name_______________________________________________________________________________ Member ID #____________________

Address______________________________________________________________________________FL License # ____________________

City__________________________________________________ State__________________________ Zip____________________________

Total amount of order____________

Total Fees Enclosed ____________

Account Number_________________________________________________________________ Exp Date_____________________________

Billing Address_______________________________________________________________________________________________________

City______________________________________ State ________________________________ Zip_________________________________

Signature________________________________________________________ Daytime #__________________________________________

You can also order Continuing Education Modules online at www.americanmedtech.org

� Visa � Master Card � American Express � Discover � Check (US funds only) payable to:

American Medical Technologists

10700 W. Higgins Road, Suite 150

Rosemont, Illinois 60018

847-823-5169

Rev. 8/12


AbstractOsteosarcoma is a highly malignant tumor of

bone, mostly affecting children and adolescent age groups. This tumor is characterized by com-plex genetic changes, which are supposed to be involved in osteosarcoma tumorigenesis. Sec-ondary osteosarcoma also can develop as se-quences of bone abnormalities such as fibrous dysplasia, Paget’s disease, and osteomyelitis. The peak incidence of osteosarcoma occurs be-tween 1 - 22 years old groups with a smaller peak after age 50. Osteosarcoma typically arises at the growth plate (Metaphysis) of the long bones, and it is mostly high grade and tends to develop pul-monary metastasis. In spite of clinical manage-ment, the patients with metastasis or even with local disease have a poor prognosis. In addition, there is no advance treatment for osteosarcoma, only cytotoxic therapy and limb amputation. This review focuses on the existing data which indicate that osteosarcoma tumors display a broad range of genetic alterations, including loss, deletion, and rearrangements of chromo-somal regions. There has evolved some inactiva-tion of tumor suppressor genes and deregulation of some signalling pathways. Certain studies found that p53 and pRb mutations are most com-monly reported in osteosarcoma patients and also reported in the animal model cell lines.

This review focuses on the importance of ani-mal models because of the difficulty to obtain human samples with different classifications of the disease, and also a limited number of clini-cally related with biological insights. Further-more, human tissue is often taken from chemo-therapy patients who then have the problems of a cellular tumor stroma and tumor necrosis in-duced by the chemotherapy. For these reasons, relevant animal models are essential to provide an investigational tool for the study of osteosar-coma. This review provides the comparison of

human, canine, and murine models in particular, and feline and rabbit in general including inci-dence, risk factors, clinical presentation and di-agnosis, genetic abnormalities, biological behav-iors and prognostic factors. Finally, this review suggests osteosarcoma should be considered as a dedifferentiated disease caused by genetic changes that interrupted osteoblast differentia-tion from mesenchymal stem cells. Thus, under-standing the molecular pathology of osteosarco-ma could lead to the development of better diagnosis and prognostic markers, as well as therapeutic agents for osteosarcoma patients.

IntroductionOsteosarcoma (OS) is the most primary malig-

nant tumor of bone. It occurs most often in ado-lescence and affects the long bones most com-monly. While relatively rare, with approximately 170 cases of bone cancer per annum in Australia (Australasian Association of Cancer Registries 2006), and approximately 600 patients develop-ing OS each year in the United States1, it is still the most common pediatric non-hematological sarcoma of bone and the 5th most common ma-lignancy of adolescence. Over the past two de-cades, even with significant advances in oncolo-gy fields, little has changed in the management of this disease. Treatment still involves the use of high-dose cytotoxic chemotherapy and tumor resection, with 5-year survival rate between 65% - 75% for localized disease and 20% with me-tastasis patients2. Cure rates for patients with metastatic or relapsed disease are poor (<20% survival). With such statistics, studies address-ing the mechanisms associated with the initia-tion and maintenance of OS may provide valu-able insights into cancer biology and may provide new avenues for therapeutic targeting. Furthermore, identifying the oncogenes which induce the aggressive behavior of OS will poten-

Haider Saddam Qasim, MT(AMT), Clinical Laboratory Consultant/Medical Scientist (Anatomical Pathology)B.M.L.Sc., L.I.B.M.S., P.M.I.A.C., F.A.S.C.P., C.L.C (NCA). M.L.S. (NZ)

The Role of Animal Models in Determining the Genetic Bases of OsteosarcomaHaider Saddam Qasim

Article 400 2 Clock Hours


tially assist the development of valuable diagnos-tic indicators and therapeutic agents. Extensive researches over the years have analysed genes in primary human OS and metastatic lung lesions and identified genes involved in metastatic poten-cy3. Studies have indicated that genetic profiling may assist to understand the pathogenesis of OS and whether or not it is related to familial syn-dromes.

There are four known causes of OS: genetic mu-tations, chemical compounds, radiation, and envi-ronmental agents2. Some genes are identified by mutated or over expressed in tumor cells and me-tastasis. However, the contribution of these mutat-ed genes to initiate in OS is still unclear4. This re-view will focus on the genetic profiling of OS, especially that involved with human, canine and murine models. The importance of canine and mu-rine models of OS has been demonstrated by a number of genetic similarities found in humans. The similarities between canine and human OS are in biological behavior. Further, distribution in the tibial and knee joints occurs naturally in both spe-cies4. Comparisons have been established of human and murine models of the genetic muta-tions (deletions or overexpressed genes), and sim-ilarities noted in tumorigenesis, aggressiveness and metastatic behavior4. The treatment of OS is unchanged over the last 30 years, with only surgi-cal resection and adjuvant chemotherapy being the two available treatments5. The successful treatment of OS will depend on the identification of genes responsible to induce the OS. This will allow the development of gene therapy and immu-notherapy against the tumor aggressiveness.5.

Incidence of OS

OS is highly prevalent in children and in young adults ages up to 21 years old6,5 (see table 1). There have been reported cases of OS in elderly individ-uals (56 - 80 years old) in association with Paget’s disease of the bone7. Paget’s disease is a disorder of the normal bone structure, formation and re-modelling process, leading to the development of various complications including bone pain, defor-mity, pathological fracture and arthritis8. There is no evidence of gender preference in OS9. However, overall male predominance in gender predilection with male: female ratio 1.5: 110. In all cases of OS, the sites affected by the primary malignant tumor are the long bones such as the Femur, Tibia and Humerus11. In 10% - 20% of cases, the tumor occurs in head, and neck and most are located in the man-

dible or maxilla and extragnathic bones.12 (See table 1).

OS Animal Models

To investigate the pathogenesis of human OS re-quires an analysis of clinically relevant animal models. The ideal animal model should provide varying degrees of tumor growth at the primary site and of metastatic potential of human OS. Such models should be valuable as tools to investigate factors (chemical, radiation, and genetic factors) that promote or inhibit OS growth and/or metasta-sis. For example, one method is to investigate the differential gene expression patterns resulting in promotion of cell proliferation and inhibition of differentiation, and which genes could potentially promote or inhibit metastasis. Animal models can also be used to test potential new therapeutics such as anti-tumor small molecules or other tar-geted therapies13, 14. The best experimental models for preclinical cancer studies must meet several criteria: (1) determine methods which are used to induce the tumor; (2) clinical and pathological similarity with the corresponding human tumors (see table 1 and 2).

Tibial Injection Model In general, the mouse model aimed to replicate

many factors of human disease behavior, especial-ly in tumor biology, genetic expressions, causes, and response to the therapeutic agents. It reflects some of the most prominent features of the human OS, such as lung metastasis (see table 1 and 2). Moreover, the model may provide some under-standing of the complex process of development and pathogenesis of OS15, and also can be tested for many kinds of therapeutic agents in vivo, at pri-mary sites, to determine the terms of development of metastasis. This model is very helpful to image and monitor the tumor progression in living mice16.

The advantages of the mouse model to under-stand the behaviors of OS are the following17: The OS will develop after inoculation in bone, and within a short period time allow for tumor progres-sion analysis. It has the ability to spread within a short time. This model demonstrates the several marker expressions characteristic of OS in vivo (but not all in vitro), such as alkaline phosphatase (ALP), pro-B 1-collagen (COL), osteopontin (OPN), matrix Gla protein (MGP) and osteocalcin (OSN)18. OS in the mouse model displays the char-acteristics of osteoid matrix and spontaneously provides development of pulmonary metastasis,


which is the most common feature of humans OS. A study explaining the principal injection of

mouse by tumor cells, must follow different crite-ria: (1) cell lines must be extracted and prepared; the cancer cells are to be grown in culture and har-vested when sub confluent; cell suspension needs to be prepared in phosphate buffered saline and placed on ice.

OS cells were obtained from the primary culture of a spontaneous OS which arose from the distal femur of an 894-day-old female Balb/c mouse.19. (2) Mouse preparation usually uses inhaled isoflu-rane to anesthetize the mouse. Other methods can

use anesthetic injection to achieve the same ef-fect. The efficiency of anesthesia is assessed by using toe pinch. There should be no reflex when the toe is pinched. Properly position the anesthe-tized mouse, which was already shaved. Then, the cells must be injected into the long bone. The ab-domen is readied with a betadine solution. The ab-domen and surgical site are covered in a sterile fashion20. Recently, the development of a novel bioluminescence in vivo imaging technique (IVIS) allowed to monitor in real-time the labeled sub-stances in experimental animals. This technology was successfully used to study tumor progression

Table 1. Genetic Abnormalities Associated with Osteosarcoma in Human and Animal Models

Osteosarcoma Human Dog Tibial injected mouse model

Double conditional knockout mouse model Cat Rabbit

Genetic abnormalities including inherited genes

P53, Rb, MDM2, RECQ helicases, Telomere maintenance (96, 97). Inherited diseases: Li-Fraumeni (p53), Rothmund-Thomos (RTS (RecQL4)), Werner Syndromes (WRN (RecQ2)), Bloom Syndrome (BLM (RecQL3)) (53)

P53, Rb, MDM2, RECQ helicases, Telomere maintenance (2)

Conditional knock out p53 and Rb. (4)

Inactivate p53 (46) Not identified

Sporadic genetic mutation

TWIST, MYC, CDKN2A, CDKN2B, BUB3, FGFR2, PRIM1, MDM2, TSPAN31, GLI1, Rb1, TOPS3A, MAPK7, COPS3, PMP22, TP53, DCC, CHK2 (53)

Not reported Not reported Not reported Not reported Not reported

Frequency of p53 mutation 15%-30% (78) 16%-29% (2) Not applicable

100% completely floxed p53, and 30%-77.8% of heterozygous floxed p53 (4)

3.4% (46) Not reported

p53 mutation pathwayRearrangement and deletion gene reported (98)

Not reported Deleted Knock out Alteration (46) Not reported

Rb gene Disrupted Disrupted Normal Not applicable Not reported Not reported

RECQ helicases pathway

Mutation been identified with patients of Rothmund-Thomason syndrome and correlated with OS risk (99)

Not been evaluated Not reported Not reported Not reported Not reported

PTEN gene pathwayMutation in other human cancers but not examined in human OS (2)

Overexpression reported in canine OS cell lines (37)

Not reported Not reported Not reported Not reported

HER-2/erbB2 oncogeneOver expressed (100)

Over expressed in cell lines and only expressed in dog tissue samples (101)

Not reported but it would same as double conditional model

Not reported Not reported Not reported

IGF and IGF-1

Over expression and anti-apoptotic , and play a role of OS proliferation and invasiveness (102)

Over expression and anti-apoptotic , and play a role of OS proliferation and invasiveness (102)


HGF and its receptor c-Met Over expression (103)Over expression in cell lines (104)


Other oncogenesc-myc, c-fos, MET, SAS, GLI are all over expressed (105)

cH-ras, c-sis, c-myc are all over expressed (2)


Metastasis related genesEzrin (106), Annexin-2 (3), CXCR4 (107), Fas-ligand (95), COX-2 (108)

Ezrin (106), COX-2 (108)Do not use this model to monitor the metastases activities

Not reported Not reported Not reported

continued after center spread


and verify the advantages of new therapies.21, 22. A Luciferase labeled metastatic OS model has been established in vivo. Highly and poorly metastatic cell lines were originally established from an OS, spontaneously arising in a Balb/c mouse19. Meta-static cells were selected and engineered in vivo to express the reporter gene (Luciferase). Upon or-thotopic injection, the IVIS technique allows mon-itoring tumor growth and metastases. IVIS analy-sis and gene expression showed that the metastatic OS cells phenotype was maintained after propagation in culture and gene marking.16 (see table 1).

Transgenic Mouse Model A transgenic mouse contains additional or artifi-

cially introduced genetic material into other cells. There are two main methods to produce transgen-ic mouse model. The first method is pronuclear microinjection (which is isolated). After fertiliza-tion, use a fine needle to inject the foreign DNA into the large male pronucleus derived from sperm, then is implanted in a foster mother and develops into an embryo. Then the DNA tends to integrate to a random position in the genome. Therefore, the resulting mouse may be partially transgenic.23. The second method to producing the

Table 2. Clinical and Pathological Features of Osteosarcoma in Human and Animal Models

Osteosarcoma Human DogTibial injected mouse

modelDouble conditional

knockout mouse modelCat Rabbit

Grade High grade primary (2) High grade primary (109) Variable grade High grade DKO model (4) High grade (43) High grade induced by radiation (42)

Incidence1-3 cases/ mill worldwide (110)

> 8000 cases/ year registered (USA only) (109, 111)

Artificial induces OS Not recorded Uncommon, 3.1-4.9 cases/100,000 (44, 45)

Rare, but few cases have been reported (42)

Gender (1.5male:1 female) (2) Equal (2) Not reported No significance (4) Not reported Not reported

Age75% of cases between 10-25 years of age. 10%over 50s with secondary OS (2)

Occur commonly middle-age to older ages, with median 7 years, however, many cases reported OS between 18-24 months (30)

Best to inject as young as possible (4-5 weeks age)

OS can induced artificially by chemical injection with 18 months old mice and 21 days OS detected after injection (4)

Average age between 8-10 years (27, 48)

Median age reported 6 years old (42)

Location in boneMetaphysis of the long bone (2)

Metaphysis of long bone (2) Not reported Metaphysis of long bone (after injection)

Metaphysis of long bone (46)

Metaphysis of long bone (42)

Distribution50% knee joint (10), 25% femur, 15% tibia, 10% humerus

75% occur in appendicular skeleton (14-15), forelimbs predominantly. Distal radius 18%, proximal humerus 7% (112)

Only in tibia Jaw, head, followed by the hind leg and hind hip, and ribs and vertebra (4)

Appendicular, axial & extraskeletal sites (O6), and subcutaneous OS (47)

Distal scapula to the diaphysis of humerus (42)

Location of metastases Lung (2) Lung (2) Lung Lung (4) Adjacent soft tissue (48)

Sp read th rough adjacent bone and there is no evidence of metastasis to the lung (42)

Aetiology Unknown Unknown Injected OS cell lines Unknown Unknown Unknown

Experimental induced OS

Chemical agents: Beryllium, Viruses: FBJ-virus, Irradiation induces: secondary OS, Electrical burn, and Trauma (96)

Mutagenic effects of ionizing radiation (113) Multiple minor trauma (114), and Genetic alterations (87)

Injected OS cell lines from double conditional knockout model cell lines

Genetic manipulation, double conditional knockout of p53, pRb genes Chemical agents: Radiation agents (4)

Not reported Radiation (42)

Clinical featuresIntermittent pain. Swelling at involved site (2)

Acute to chronic lameness, swelling of the affected limb (2)

Same Same (24) Same (48) Same (42)

Radiological Diagnosis

Aggressive, metaphyseal lesions of the long bone, and the tumour ranged from lytic to blastic OS (115)

Aggressive, metaphyseal lesions of the long bone, and the tumour ranged from lytic to blastic (2)

Using luciferare imaging and micro CT

Aggressive destructive growth toward the joint with periosteal new bone formation at the site of injection (24)

Aggressive, destruct the cortex of the bone and involve surrounding soft tissues (48)

Destructive lesion at the bone from distal scapu la to the diaphysis of the h u m e r u s ( n o surrounding tissue involved) (42, 116)

HistologyMalignant spindle cells, production of osteoid matrix by tumour cells (117)

Malignant spindle cells, production of osteoid matrix by the tumour cells (2)

similar with double conditional model

Osteoid, fibroblastic tumour, malignant spindle cells, abnormal mitotic, multi-nucleated giant cells (4)

Malignant mesenchymal spindle cel ls, which produced osteoid (48)

Round to polyhedral neoplastic cells, round to oval nuclei, high c e l l u l a r , pleomorphism, large number of multi-nucleated giant (116)

continued from page 52


transgenic mouse is by introducing the DNA into embryonic stem cells; then the embryonic stem cells need a host embryo to develop and are isolat-ed from a mouse with different color fur. The im-planted embryonic stem cells will colonize a host embryo, and the resultant mouse — in the first generation being chimera (mixture brown and black colors) — the resulting sperms will carry the features of the original injected DNA. Then when these transgenic sperms fertilize a normal egg, a transgenic mouse will produce with the same for-eign DNA in whole body cells23.

The most modern model was a genetically engi-neered technique performed on the mouse based on osteoblast-restricted deletion of p53 and pRb. As a result, the OS development is dependent on loss of p53 and Rb, revealing a dominance of p53 mutation in the development of OS. This model is a crucial to addressing the molecular genetics of OS comparable with human, and valuable for de-veloping therapeutic agents.4 (see table 2). A study was done in Koch Institute for Integrative Cancer Research in MIT to successfully generate a double conditional knockout model (Osx-Cre+p53fl/flpRbfl/fl double conditional knockout mice). The combined deletion of both genes (p35 and pRb) gave completely penetrant OS model and developed many defining charac-ters similar to human OS, such as gene expres-sion signatures, histological characters and meta-static potential1. To generate the double conditional knockout model, the same study used mice that carried three alleles, which are the con-ditional allele of pRb, p53 and Osx1-GFP-Cre transgene24. Cre recombinase expression is con-trolled by Osterix1 (Osx1) promoter (Osx is a gene which differentially expresses at the earlier stages of osteoblast)25. Crossing Rb+/c, Cre+, p53+/c, Cre+ or Rb+/cp53+/c, Cre+ males with Rbc/c, p53c/c or Rbc/c, p53c/c females will gener-ate animals with all possibilities of combination between p53 and pRb with or without Cre. It was confirmed that Cre only expressed in specific os-teoblast and not other mesenchymal cells, and also confirmed that Cre catalytic activity only to the conditional pRb and p53 alleles in bone. The breeding between fluxed p53 and pRb with pres-ence of Cre expression will develop a genotype highly predisposed to develop OS with short la-tency and complete penetrance, and the metasta-ses are commonly seen in lung and liver24. The double conditional knockout model (DKO) dis-played typical characters of human OS such as

histological features which showed destruction of the bone cortex, ossified spicules in the tumor which will be arised outside the periosteum (4). In addition, DKO model provides similarities in X-ray analysis with human OS by revealed sun-burst pattern which indicate osteoid tissue (osse-ous before calcification stage)24.

Similarities Between Double Conditional Knockout Model (fluxed p53 and pRb) and Tibial Injected Model

A study24 addressed a number of similarity char-acters between DKO model and tibial injected model by doing the following: (1) Harvested the OS tumor from the DKO mouse model. (2) Me-chanically disaggregated tumor cells. (3) Placed those cells into the tissue culture. (4) After the tumor cells were grown in tissue culture, the cultured cells were injected into the nude immuno-compromised mouse intravenously and subcutaneously. (5) After 50-100 days, the tumor arised. (6) A biopsy was taken from the arised le-sion. H & E staining, Sirius Red stain, and Alizarin Red stain indicated that the arised lesion tumor closely resembled the parental OS which was poorly differentiated to undifferentiated. Further-more, the subcutaneous injected tumor was highly invasive and metastasized into the lung and liver24.

Canine ModelThe clinicopathologic features of the canine OS

closely approximates the morphology and biology behaviors of human OSs26 (see table 1) . Further-more, the clinical staging system of OS in canine is highly similar in human OS, and because of the short life span of the dog, it is very easy to mani-fest the neoplasm within a short period. On the other hand, canine model clearly demonstrates that therapeutic results can be made within a shorter period time than human. For example, one year in OS survival in the dog is roughly equivalent to 4 - 5 years in human27, 28 (Table 1) .

The epidemiological studies in the USA (Ala-bama and San Francisco universities) show the rate incidence of bone tumor is 7.9/100,000 dogs, whereas the incidence of bone tumors is 1.1/100,000 human29.

The fibrosarcomatous subtype is associated with a relatively favorable prognosis in both spe-cies30. The best comparison in age incidence be-tween human and dog OS, formulated by Lebeau, is 12 years old in human is equivalent to 1 year in dog; that is mean. With the OS diagnosed in a 24


old human, it will be equivalent to 2 years of OS diagnosed in dog28.

Age and site predilection are different. Most of the affected dogs are older animals, whereas mainly adolescents are affected in human. This feature represents the most difference between dog and human of age incidence29. The dog limb bones and the knee region in the human are the sites involved. The varying localizations in man and dog, cat, cow, horse, and rodent might be as-sociated with the stress of weight burden (radius, humerus, tibia and femur)30. Males have slightly more incidence than females in man and dog31, 32. 77% of canine OS arises in the long bones, and only 23% involve flat bones33. The majority sites of OS affected the dog forelimbs specifically in the prox-imal humerus and distal radius. Human OSs are arising in irradiated bone, in bone affected by Pag-et’s disease, and in skeletal tumors (multiple en-chondromas, multiple osteochondromas, fibrous dysplasia, and chondorsarcomas)34, 35. The majority of clinical findings in canine OS are lameness and swelling, and have been observed within six months. The affected areas include a hard fusi-form and painless swelling, with a severe case lymphedema at the distal side of tumor. The clini-cal pulmonary symptoms are not initially meta-static, but commonly develop the hypertrophic pulmonary osteoarthropathy at the late stage of OS metastasis33 (see table 1). The histological fea-tures of canine OS are highly identical with human OS, which is character finding of osteoid by the sarcoma cells and varying amount of chondroid and collagen33, 36. A study of 144 untreated dogs with OS indicated 49 dogs diagnose with metasta-sis, and 44 involve the lungs. The measurement of metastasis correlated with the tumor diameter, volume, and extension from the original site30 (see table 1). Canine OS (OS) cell lines arise from mu-tations that directly inactivate the tumor suppres-sor genes p53 and retinoblastoma. Another gene such as PTEN gene (phosphatase and tensin ho-molog consider as a tumor suppressor gene through the action of its phosphatase protein product) is mutated in many human cancers37. PTEN is mutated or down regulated in a high per-centage of canine OS tumor cells37 (see table 2). Mutations of the p53 gene in canine OS were screened for mutations of the complete p53 gene using PCR. The mutated gene was analyzed by sin-gle-strand conformational polymorphism. The clinical outcome of dogs with p53-mutated OS was compared to dog OS without a mutation. After

complete surgical excision of the primary tumor, which is demonstrated by the p53 mutation com-mon in canine OS, there may be may prognostic value that suggests the mutations of the p53 gene might be influenced by survival rate and it should be considered when evaluating the canine OS38. Some case reports diagnosed the metastasis of ap-pendicular OS. The appendicular OS metastasis extended to the anterior limbs. Some clinical find-ings showed detectable metastases to the eye be-fore systemic organ metastases39 (see table 1). However, there are no specific oncogenes that contribute to the pathogenesis of canine OS (OS). The over expresses the sis oncogene that encodes the platelet-derived growth factor (PDGF)-B. All of canine OS cell lines tested contained PDGF re-ceptors. These cell lines are capable of responding to PDGF given the importance of PDGF in pro-moting cell proliferation, migration, and cell sur-vival. Therefore, the activation of the sis oncogene potentially contributes to the pathogenesis of a subset of canine OS040 (see table 2). The study of isolated and characterized cancer stem cells from canine OS suggests that cancer is a stem cell dis-ease37. One of human OS cell line and three canine cell lines were cultivated in non-adherent media by used serum-starved and semi-solid medias. The grower colonies from all cell lines were identified under these conditions and were characterized by using molecular and cytochemical techniques for embryonic stem cell markers. Similarities be-tween human and canine OS cell lines support the dog as a model for human disease.41. A compara-tive study of OS may lead to new etiologic and pathogenetic concepts.

Advantages of Canine Model For OS OS in canine28 is more common than human.

The clinical and diagnosis of canine OS is very easily recognized. Both humans and dogs share the same environment and similar causative agents; that should help to determine the envi-ronmental factors that might induce OS. The large size of dogs allows enough sample collec-tion of blood and tissue samples. However, this characteristic does not present in the murine model. Unlike in dogs and humans, the murine model has no osteonal contexture, for example, a harversian canal; that is mean. The development of OS in dog osteonal bones is more closely aligned to human OS. Close similarity in the clin-ical, radiological and histological features of dog OS with human OS may give invaluable informa-


tion about the etiological, epidemiological, im-munological, and therapeutic features of human OS.

Rabbit Model Osteosarcoma occurring in domestic rabbit is

rare, but a few cases have been reported42, and it is highly induced by radiation. The median age reported is 6 years (see table 1). The clinical fea-tures of the rabbit are swelling at the site of le-sion, with fever and lameness, and increase in the ALP42. Many cases reported marked proliferative and destructive lesions of bone at the diaphysis of the humerus and involve joint space (see the table 1). Histological finding are similar among human, dog and rabbit which are round-to-poly-hedral neoplastic cells that resembled histocytes; round-oval-nuclei, highly mitosis, pleomorphism; atypical nuclei and cytoplasm are predominant; and a large number of muti-nucleated giant cells (see table 1). Radiological diagnosis demon-strates the destructive lesion at the bone from distal scapula to the diaphysis of the humerus and no surrounding tissue involved (see table 1). However, no classification of osteosarcoma has been established. Furthermore, there are no re-ports of osteosarcoma metastasis to the lung in rabbit42. In addition, it is seldom that osteosarco-ma involves the joint spaces or surrounding soft tissues42. Finally, there are no reports referring to the genetic involvement in rabbit osteosarcoma42.

Feline Model Feline osteosarcoma is relatively infrequent,

but it is the most common malignancy of the bone tumors in the cat43. The incidence has been reported of 3.1-4.9 per 100.000 cases44, 45. The av-erage age occurrence is between 8-10 years27 (see table 1). There is no report that recognizes the gender preference of osteosarcoma in feline model46. Feline osteosarcoma, located in the ap-pendicular, axial of the long bones and extraskel-etal sites, accounted for 40% of all feline osteo-sarcoma47. Also, the extraskeletal osteosarcoma can be induced by injection sites. However, it has a poorer prognosis than the skeletal osteo-sarcoma46, 47. Low metastasis rate of only 5-10% is reported in adjacent soft tissues46 (see table 1). The histological characteristics of feline skeletal osteosarcoma are not significantly different from the canine osteosarcoma model46, 48. The X-ray diagnos revealed the aggressiveness, de-struction in the cortex of the bone and involved

surrounding soft tissue46. Genetically, the muta-tion of p53 in feline cases of osteosarcoma has been identified (see table 2). Data confirmed that osteosarcoma in the feline model is caused by the mutation in the coding regions of p53 gene46,

49. A study used the IHC (Immunohistochemistry) analysis of p53 in feline neoplasia (46). The IHC staining analysis demonstrate p53 mutation in four cases of osteosarcoma out of 77 cases from different tumors, and also found the p53 inacti-vate mutation in lymphoma, mammary carcino-ma and osteosarcoma, and the p53 mutated over-expressed in squamous cell carcinoma46. This study suggests the mechanism of tumorigenesis may be similar in both human and feline.e. Fur-thermore, all the 77 feline tumour samples demonstrate the immunostaning only appear at the nucleus and no signals at the cytoplasm, which is provide strong evidence to support p53 abnormalities in the pathogenesis of different types feline neoplasia46.

Summary of animal modelsMouse, dog, cat and rabbit are considered as

mammalian animal models, serving in use for research and investigation of human disease, for the purposes of better understanding of the diseases without added any risk to humans during the process. OS is more common in the canine model than in the human model. Unlike the murine model, the OS is spontaneously induced in the canine model without genetic manipulation. Murine and canine models share many similarities in clinical and diagnostic features, histologically and in aggressiveness, as well as some oncogenes overexpressed in OS. The genetic engineering techniques had been performed on osteoblast - knockout of p53 and pRb with using Osx1 - Cre, which is called double conditional knockout (DKO). The transgenic model has early developed OS with high penetrance and short latency. In contrast, the interrupted overexpression of p35 and pRb pathways results the canine OS tumorigenesis. The osteosarcoma in the feline model reported by mutation in the coding regions of p53 gene. However, there are no data reported on genes expressed in rabbit osteosarcoma. Therefore, the animal models replicate many facts of human OS mechanism behavior, metastasis and pathogenesis; in addition, these models are helpful to monitor the progression of disease.


The Genetic Basis Of Osteosarcoma (see table 1)

No specific translocations or genetic abnormali-ties in humans have been identified to cause OS;14, 50-54 However, approximately 70% of OS tumor cells display the contribution of some genetic vari-ation, which leads to cytogenic abnormalities54, 55.

RetinoblastomaAll OS cell line results indicate that they are de-

ficient of Rb pathways56; retinoblastoma is a tumor suppressor gene. Rb family consists of Rb, P107, and P130, and they produce proteins which regulate cell proliferation and differentiation57-59. Normally, the pRb gene regulates the cell cycle by controlling the activities of a number of E2F fam-ily. A group of activator and suppressor coding genes of transcriptional factors in high eukaryotic cells participate in cell cycle regulation and DNA synthesis60. Many reports recognize the absence of Rb pathways, the Rb mRNA levels will increase to compensate the loss or inactivation of Rb pro-tein.61, 62. The patients exhibitory pRb mutations have approximate 500 folds higher incidence of OS than the general population. Furthermore, the pRb mutations have been detected in 70% of all adolescent diagnosed with primary OS63-72. More-over, the pRb protein interacts with Runx2, which results the complex transcriptional activities in-volved the late stage of osteoblast marker osteo-calcin73 Thus, the loss of pRb can suppress the terminal stage of osteogenic differentiation in vitro74. However, a heterozygous Rb mutation pro-duces a different type of tumour, but no OS73.

p53P53 is a tumor suppressor protein which is en-

coded via TP53 gene. This gene plays a major role in osteoblast differentiation and bone devel-opment. P53 rearrangement and/or deletion of both alleles are common in human OS. P53 is ac-tivated in response of various stimuli which in-cluding DNA damage or promote nucleotide im-balance. A study by A.J. Levine was found the P21 and mdm2 inductive are considered to be one of defective properties of canine OS cell lines.75 In another study was found the loss of heterozygosity at the Rb1 locus is present in ap-proximately 60% to 70% of OS tumors67. An ab-sence of the p53 oncogene has been found in the early and metastatic stages of OS76. Other reports identified the chromosomal location of 17p13, as abnormal in OS, and that is where p53 is locat-

ed55, 77. However, p53 mutation status is not asso-ciated with the stage of OS tumor and/or metas-tasis78. Many studies demonstrate the p53 has ability to modulate the differentiation of osteo-blasts, as well as play an important role in the bone development. Thus, the p53 deficient mice has accelerated osteoblast differentiation and in-creased bone density79. The hyperactivation of p53 is induced by deleting mdm2 (p53 inhibitor), which leads to suppressed osteoblast differentia-tion by inhibited Runx2 (a bone specific tran-scriptional factor80.

The deletion of p53 from mesenchymal stem cells or from osteoblast precursors in vitro, that promotes transcriptional changes associated with early stages of osteogenesis but stops or im-pair at the end stage of differentiation to mature osteocyte76. In addition, the reduced levels of p53 mRNA are possibly resulted from inhibition of transcription or alteration of mRNA degradation, which is lead to compromised p53 activities in tu-mour cells56, 81. Many researches indicate the p53 is overexpressed in 84% from tumour cells de-rived from appendicular bones location, and in 56% from tumour cells derived from axial bones location75, 81.

The Relation Mechanism Between p53 and pRb Genes

Through studies performed by Nigg and Weinberg, it was found that any mutation or inac-tivation of Rb protein or response of growth in-hibitory signals, the cyclin-dependant kinases (cdk) 4 and 6 inhibitors block the pRb pathway by decrease the inhibitory activities of pRb or de-regulate cyclin D60, 2. On the other hand, a study found that the p53 activity induced during any cell stress or DNA damage; by transcript mdm2 and cki p21 (is a protein encoded by CDK inhibi-tors and mdm2 is a negative regulator of p53 pro-tein stability), both block of Rb phosphorylation61,

62, 74, 83. In addition, the p53 participate in pro-grammed apoptosis program of the cell in re-sponse to DNA damage, hypoxia, or chemothera-peutic agents84, 85 and because of the tumour cells have activated oncogenes, the p53 and pRb path-ways are already inactivated, these features give more resistance against apoptosis and give a chance to accumulate additional mutations which contribute to tumour progression and ag-gressiveness86. A study used IHC techniques to demonstrate the p53 is overexpressed in high percentage in canine OS cell lines87, 88, and conse-


quently there are missing specific fragments in the pRb, for example missing 3.7kb Rb specific sites which are presented in the control bone.

Inactivation p53 and pRb in murine OS cell lines

The mutation of pRb and p53 tumour suppres-sors is associated with OS development, using the transgenic mouse strain with inactivated p53 and pRb in osteoblast precursors. The result of double conditional knockout mouse (DKO) of p53 and Rb has early developed onset of OS with complete penetrance24, DKO (pRb c/c; p53c/c) with the Osx1-Cre transgene (Cre recombinase is an enzyme that delete a gene between the two targeted sequences called loxP. Osx1 is a gene which only expressed in earlier stages of osteo-blast development), produces a transgenic mice with complete OS penetrance and short latency, whereas die as early as 4 months age89. DKO OS cell line in mice share many characters of human OS such as90; predisposition to develop the tu-mour within femur, knee joint and lower femur and upper tibia, when are considered to be active bone growth and repair sites. Cellular composi-tion is similar to human and both species and share high incidence of pulmonary metastases is a major cause of death. Both sporadic and famil-iar OS predispose the inactivation of p53 and pRb play a key role in development of OS and other tumours such as neuroendocrine tumours, and hibernomas53, 91

Inactivation p53 and pRb in canine OS cell lines

Canine model of OS has been used to study the cancer biology and treatment; however the pathogenesis of canine OS is still unknown92, the similarity characteristics between human and ca-nine OS are the same aggressiveness93, highly metastatic tumour, similar histology94, difficult to manage clinically, because the aggressive biolog-ical behaviour of this tumour, and same location tumour distributed of the long bones. Further-more, both species are associated with abnor-malities of p53 and pRb genes in the OS. On the other hand, there are different characters be-tween human and Canine OS including that it is a common tumour in dogs but rare with human95. OS generally occur in older age in dogs whereas

OS occur in adolescent age group in human95.

The Comparison Between p53 and pRb of Mu-rine and Canine OS Cell Lines

All the results obtained from the canine OS cell lines underscored the inactivation of p53 and pRb in canine OS tumorigenesis92. The p53 mRNA is ei-ther overexpressed or not expressed in canine cell lines OS2. The pRb mRNA is overexpressed with high levels in all canine cell lines OS. P53 and Rb proteins are overexpressed in high percentage of canine OS2, whereas in all DKO mouse OS cell lines, the p53 and pRb genes are absolutely knock-out24.

Common Methods are Used for Osteosarcoma Research

A number of developing technologies continue to provide us a comprehensive assessment to un-derstand more about the tumor behaviors. Re-cently, massive studies have been done of the mo-lecular genomic sequences of osteosarcoma by using techniques of genomic array, FISH, and ISH. To understand the mRNA expression of osteosar-coma and also in many tumors, the RT-PCR is the best way for these purposes. Western blot (WB) experiments have been used for more under-standing of protein expression which resulted from the gene or oncogene expression. Flow Cy-tometry (FC) is a very widely used method to un-derstand the tumor cell surface expression and to determine which CDs or markers would differen-tiate a tumor from others. It is a very valuable method to understand the tumor cell expressions in metastasis stages in vitro. Remarkably, Immu-nohistochemistry (IHC) is the best method to lo-calize the proteins expression in the tumor tissue sample and it will provide important information of OS tumorogenesis and aggressiveness. Finally, the PCNA (Proliferating Cell Nuclear Antigen) is a valuable method to measure the invasiveness and aggressiveness of malignancies including all kinds of osteosarcoma.

Summary of Genetic Bases of OS

Genes expression profiles will provide valuable information about cancer biology and behaviors. It can be used for developing biomarkers to identify consequent genes and be involved in a specific function of tumor molecular biology. On the other


hand, the initial mutations have been linked to ac-tivate oncogenes or inactivate the tumour sup-pressor genes, which can trigger certain subse-quent events leading to cellular transformation and tumor formation. Finally, the identification of these genes might increase the understanding of how osteosarcoma develops and progresses to malignancy as well as the tumorigenesis and me-tastasis behaviors. Conclusions

This review describes the importance of submis-sive animal models of human disease which will offer many opportunities to advance our knowl-edge and understanding of cancer biology and ge-netics of human cancer. The modern genetic engi-neered mouse (DKO) model provides many similarities to human OS characteristics. On the other side, a number of studies demonstrate the similarity between the tibial injected model and DKO model. In comparative oncology, canine model of OS has great potential with characteris-tics of human OS, especially in histology, clinical relevant, biological advances, pre- and post- clini-cal treatment evaluations. Interestingly, many genes are expressed and/or overexpressed in OS common among human, mouse, and canine mod-els. Therefore, the study of genes expression and the mechanisms are involved to initiate OS are very important to understanding the tumorigene-sis, invasiveness and aggressiveness of this tumor. In other words, the development and utilization of murine and canine models faithfully reflect the OS behaviors in humans, which is considered a signif-icant promise to enhance our understanding of OS genetic basis. Finally, the advance in diagnostic methods and therapeutic agents will be enhanced by identifying the genes that contribute to OS ag-gressiveness, and identifying the genes involved in primary OS and other genes that contribute to po-tential metastasis.

References1. Janeway KA, Walkley CR. Modeling human osteosarcoma in

the mouse: From bedside to bench. Bone. Nov;47(5):859-65.2. Mueller F, Fuchs B, Kaser-Hotz B. Comparative biology of

human and canine osteosarcoma. Anticancer Res. 2007 Jan-Feb;27(1A):155-64.

3. Gillette JM, Chan DC, Nielsen-Preiss SM. Annexin 2 expres-sion is reduced in human osteosarcoma metastases. J Cell Bio-chem. 2004 Jul 1;92(4):820-32.

4. Walkley CR, Qudsi R, Sankaran VG, Perry JA, Gostissa M, Roth SI, et al. Conditional mouse osteosarcoma, dependent on

p53 loss and potentiated by loss of Rb, mimics the human dis-ease. Genes Dev. 2008;22(12):1662-76.

5. Caudill JS, Arndt CA. Diagnosis and management of bone ma-lignancy in adolescence. Adolesc Med State Art Rev. 2007;18(1):62-78, ix.

6. Estrada-Villasenor E, Delgado Cedillo EA, Rico Martinez G. [Frequency of bone neoplasms in children]. Acta Ortop Mex. 2008 Jul-Aug;22(4):238-42.

7. Longhi A, Errani C, Gonzales-Arabio D, Ferrari C, Mercuri M. Osteosarcoma in patients older than 65 years. J Clin Oncol. 2008 Nov 20;26(33):5368-73.

8. van Staa TP, Selby P, Leufkens HG, Lyles K, Sprafka JM, Coo-per C. Incidence and natural history of Paget’s disease of bone in England and Wales. J Bone Miner Res. 2002 Mar;17(3):465-71.

9. Jasnau S, Meyer U, Potratz J, Jundt G, Kevric M, Joos UK, et al. Craniofacial osteosarcoma Experience of the cooperative German-Austrian-Swiss osteosarcoma study group. Oral Oncol. 2008;44(3):286-94.

10. Sim FH, Edmonson JH, Wold LE. Soft-tissue sarcomas. Future perspectives. Clin Orthop Relat Res. 1993 Apr(289):106-12.

11. Baena-Ocampo Ldel C, Ramirez-Perez E, Linares-Gonzalez LM, Delgado-Chavez R. Epidemiology of bone tumors in Mex-ico City: retrospective clinicopathologic study of 566 patients at a referral institution. Ann Diagn Pathol. 2009 Feb;13(1):16-21.

12. Fernandes R, Nikitakis NG, Pazoki A, Ord RA. Osteogenic sar-coma of the jaw: a 10-year experience. J Oral Maxillofac Surg. 2007 Jul;65(7):1286-91.

13. Luo X, Chen J, Song WX, Tang N, Luo J, Deng ZL, et al. Os-teogenic BMPs promote tumor growth of human osteosarco-mas that harbor differentiation defects. Lab Invest. 2008 Dec;88(12):1264-77.

14. Haydon RC, Luu HH, He TC. Osteosarcoma and osteoblastic differentiation: a new perspective on oncogenesis. Clin Orthop Relat Res. 2007 Jan;454:237-46.

15. Hann B, Balmain A. Building ’validated’ mouse models of human cancer. Curr Opin Cell Biol. 2001 Dec;13(6):778-84.

16. Miretti S, Roato I, Taulli R, Ponzetto C, Cilli M, Olivero M, et al. A mouse model of pulmonary metastasis from spontaneous osteosarcoma monitored in vivo by Luciferase imaging. PLoS One. 2008;3(3):e1828.

17. Ek ET, Dass CR, Choong PF. Commonly used mouse models of osteosarcoma. Crit Rev Oncol Hematol. 2006 Oct;60(1):1-8.

18. Zhou H, Choong P, McCarthy R, Chou ST, Martin TJ, Ng KW. In situ hybridization to show sequential expression of osteo-blast gene markers during bone formation in vivo. J Bone Miner Res. 1994 Sep;9(9):1489-99.

19. Schmidt J, Strauss GP, Schon A, Luz A, Murray AB, Melchiori A, et al. Establishment and characterization of osteogenic cell lines from a spontaneous murine osteosarcoma. Differentiation. 1988 Dec;39(3):151-60.

20. Tseng W, Leong X, Engleman E. Orthotopic mouse model of colorectal cancer. J Vis Exp. 2007(10):484.

21. Gomes CM, Welling M, Que I, Henriquez NV, van der Pluijm G, Romeo S, et al. Functional imaging of multidrug resistance in an orthotopic model of osteosarcoma using 99mTc-sestami-bi. Eur J Nucl Med Mol Imaging. 2007 Nov;34(11):1793-803.

22. Deroose CM, De A, Loening AM, Chow PL, Ray P, Chatziio-annou AF, et al. Multimodality imaging of tumor xenografts and metastases in mice with combined small-animal PET, small-animal CT, and bioluminescence imaging. J Nucl Med. 2007 Feb;48(2):295-303.

23. Davey RA, MacLean HE. Current and future approaches using genetically modified mice in endocrine research. Am J Physiol


Endocrinol Metab. 2006 Sep;291(3):E429-38.24. Berman SD, Calo E, Landman AS, Danielian PS, Miller ES,

West JC, et al. Metastatic osteosarcoma induced by inactivation of Rb and p53 in the osteoblast lineage. Proc Natl Acad Sci U S A. 2008 Aug 19;105(33):11851-6.

25. Cao Y, Jia SF, Chakravarty G, de Crombrugghe B, Kleinerman ES. The osterix transcription factor down-regulates interleu-kin-1 alpha expression in mouse osteosarcoma cells. Mol Can-cer Res. 2008 Jan;6(1):119-26.

26. Nielsen SW. Comparative pathology of bone tumors in animals, with particular emphasis on the dog. Recent Results Cancer Res. 1976(54):3-16.

27. Dorn CR, Taylor DO, Schneider R, Hibbard HH, Klauber MR. Survey of animal neoplasms in Alameda and Contra Costa Counties, California. II. Cancer morbidity in dogs and cats from Alameda County. J Natl Cancer Inst. 1968 Feb;40(2):307-18.

28. Brodey RS. The use of naturally occurring cancer in domestic animals for research into human cancer: general considerations and a review of canine skeletal osteosarcoma. Yale J Biol Med. 1979 Jul-Aug;52(4):345-61.

29. Dorn CR. Comparative oncology: dogs, cats, and man. Perspect Biol Med. 1972 Summer;15(4):507-19.

30. Misdorp W, Hart AA. Some prognostic and epidemiologic fac-tors in canine osteosarcoma. J Natl Cancer Inst. 1979 Mar;62(3):537-45.

31. Tjalma RA. Canine bone sarcoma: estimation of relative risk as a function of body size. J Natl Cancer Inst. 1966 Jun;36(6):1137-50.

32. Fraumeni JF, Jr. Stature and malignant tumors of bone in child-hood and adolescence. Cancer. 1967 Jun;20(6):967-73.

33. Brodey RS, Riser WH. Canine osteosarcoma. A clinicopatho-logic study of 194 cases. Clin Orthop Relat Res. 1969 Jan-Feb;62:54-64.

34. Dahlin DC. Giant-cell-bearing lesions of bone of the hands. Hand Clin. 1987 May;3(2):291-7.

35. Reiman HM, Dahlin DC. Cartilage- and bone-forming tumors of the soft tissues. Semin Diagn Pathol. 1986 Nov;3(4):288-305.

36. Ling GV, Morgan JP, Pool RR. Primary bone rumors in the dog: a combined clinical, radiographic, and histologic approach to early diagnosis. J Am Vet Med Assoc. 1974 Jul 1;165(1):55-67.

37. Levine RA, Forest T, Smith C. Tumor suppressor PTEN is mu-tated in canine osteosarcoma cell lines and tumors. Vet Pathol. 2002 May;39(3):372-8.42. Kondo H, Ishikawa M, Maeda H, Onuma M, Masuda M, Shibuya H, et al. Spontaneous osteosar-coma in a rabbit ( Oryctolagus cuniculus). Vet Pathol. 2007 Sep;44(5):691-4.

43. Negrin A, Bernardini M, Diana A, Castagnaro M. Giant cell osteosarcoma in the calvarium of a cat. Vet Pathol. 2006 Mar;43(2):179-82.

44. Bitetto WV, Patnaik AK, Schrader SC, 38. Kirpensteijn J, Kik M, Teske E, Rutteman GR. TP53 gene mu-

tations in canine osteosarcoma. Vet Surg. 2008 Jul;37(5):454-60.

39. Yoshikawa H, Nakamoto Y, Ozawa T, Dickinson RM. A dog with osteosarcoma which metastasized to the eye months be-fore metastasis to other organs. J Vet Med Sci. 2008 Aug;70(8):825-8.

40. Levine RA. Overexpression of the sis oncogene in a canine os-teosarcoma cell line. Vet Pathol. 2002 May;39(3):411-2.

41. Wilson H, Huelsmeyer M, Chun R, Young KM, Friedrichs K, Argyle DJ. Isolation and characterisation of cancer stem cells from canine osteosarcoma. Vet J. 2008 Jan;175(1):69-75.

42. Kondo H, Ishikawa M, Maeda H, Onuma M, Masuda M, Shibuya H, et al. Spontaneous osteosarcoma in a rabbit ( Oryc-

tolagus cuniculus). Vet Pathol. 2007 Sep;44(5):691-4.43. Negrin A, Bernardini M, Diana A, Castagnaro M. Giant cell

osteosarcoma in the calvarium of a cat. Vet Pathol. 2006 Mar;43(2):179-82.

44. Bitetto WV, Patnaik AK, Schrader SC, Mooney SC. Osteosarco-ma in cats: 22 cases (1974-1984). J Am Vet Med Assoc. 1987 Jan 1;190(1):91-3.

45. Liu SK, Dorfman HD, Patnaik AK. Primary and secondary bone tumours in the cat. J Small Anim Pract. 1974 Mar;15(3):141-56.

46. Nasir L, Krasner H, Argyle DJ, Williams A. Immunocytochem-ical analysis of the tumour suppressor protein (p53) in feline neoplasia. Cancer Lett. 2000 Jul 3;155(1):1-7.

47. Dhaliwal RS, Johnson TO, Kitchell BE. Primary extraskeletal hepatic osteosarcoma in a cat. J Am Vet Med Assoc. 2003 Feb 1;222(3):340-2, 16.

48. Dimopoulou M, Kirpensteijn J, Moens H, Kik M. Histologic prognosticators in feline osteosarcoma: a comparison with phe-notypically similar canine osteosarcoma. Vet Surg. 2008 Jul;37(5):466-71.

49. Mayr B, Reifinger M, Loupal G. Polymorphisms in feline tu-mour suppressor gene p53. Mutations in an osteosarcoma and a mammary carcinoma. Vet J. 1998 Jan;155(1):103-6.

50. Cormier JN, Pollock RE. Soft tissue sarcomas. CA Cancer J Clin. 2004 Mar-Apr;54(2):94-109.

51. Hansen MF. Genetic and molecular aspects of osteosarcoma. J Musculoskelet Neuronal Interact. 2002 Dec;2(6):554-60.

52. Hayden JB, Hoang BH. Osteosarcoma: basic science and clini-cal implications. Orthop Clin North Am. 2006 Jan;37(1):1-7.

53. Kansara M, Thomas DM. Molecular pathogenesis of osteosar-coma. DNA Cell Biol. 2007 Jan;26(1):1-18.

54. Marina N, Gebhardt M, Teot L, Gorlick R. Biology and thera-peutic advances for pediatric osteosarcoma. Oncologist. 2004;9(4):422-41.

55. Sandberg AA, Bridge JA. Updates on the cytogenetics and mo-lecular genetics of bone and soft tissue tumors: osteosarcoma and related tumors. Cancer Genet Cytogenet. 2003 Aug;145(1):1-30.

56. Zhu L, Zhu L, Xie E, Chang LS. Differential roles of two tan-dem E2F sites in repression of the human p107 promoter by retinoblastoma and p107 proteins. Mol Cell Biol. 1995 Jul;15(7):3552-62.

57. Cobrinik D, Lee MH, Hannon G, Mulligan G, Bronson RT, Dyson N, et al. Shared role of the pRB-related p130 and p107 proteins in limb development. Genes Dev. 1996 Jul 1;10(13):1633-44.

58. Hamel PA, Phillips RA, Muncaster M, Gallie BL. Speculations on the roles of RB1 in tissue-specific differentiation, tumor ini-tiation, and tumor progression. Faseb J. 1993 Jul;7(10):846-54.

59. Ikeda MA, Jakoi L, Nevins JR. A unique role for the Rb protein in controlling E2F accumulation during cell growth and differ-entiation. Proc Natl Acad Sci U S A. 1996 Apr 16;93(8):3215-20.

60. Weinberg RA. The retinoblastoma protein and cell cycle con-trol. Cell. 1995 May 5;81(3):323-30.

61. Aagaard L, Lukas J, Bartkova J, Kjerulff AA, Strauss M, Bartek J. Aberrations of p16Ink4 and retinoblastoma tumour-suppres-sor genes occur in distinct sub-sets of human cancer cell lines. Int J Cancer. 1995 Mar 29;61(1):115-20.

62. Bartkova J, Lukas J, Guldberg P, Alsner J, Kirkin AF, Zeuthen J, et al. The p16-cyclin D/Cdk4-pRb pathway as a functional unit frequently altered in melanoma pathogenesis. Cancer Res. 1996 Dec 1;56(23):5475-83.


63. Castresana JS, Rubio MP, Gomez L, Kreicbergs A, Zetterberg A, Barrios C. Detection of TP53 gene mutations in human sar-comas. Eur J Cancer. 1995;31A(5):735-8.

64. Clarke AR, Maandag ER, van Roon M, van der Lugt NM, van der Valk M, Hooper ML, et al. Requirement for a functional Rb-1 gene in murine development. Nature. 1992 Sep 24;359(6393):328-30.

65. Alonso J, Garcia-Miguel P, Abelairas J, Mendiola M, Pestana A. A microsatellite fluorescent method for linkage analysis in familial retinoblastoma and deletion detection at the RB1 locus in retinoblastoma and osteosarcoma. Diagn Mol Pathol. 2001 Mar;10(1):9-14.

66. Araki N, Uchida A, Kimura T, Yoshikawa H, Aoki Y, Ueda T, et al. Involvement of the retinoblastoma gene in primary osteosar-comas and other bone and soft-tissue tumors. Clin Orthop Relat Res. 1991 Sep(270):271-7.

67. Belchis DA, Meece CA, Benko FA, Rogan PK, Williams RA, Gocke CD. Loss of heterozygosity and microsatellite instability at the retinoblastoma locus in osteosarcomas. Diagn Mol Pathol. 1996 Sep;5(3):214-9.

68. Benassi MS, Molendini L, Gamberi G, Ragazzini P, Sollazzo MR, Merli M, et al. Alteration of pRb/p16/cdk4 regulation in human osteosarcoma. Int J Cancer. 1999 Oct 22;84(5):489-93.

69. Miller CW, Aslo A, Won A, Tan M, Lampkin B, Koeffler HP. Alterations of the p53, Rb and MDM2 genes in osteosarcoma. J Cancer Res Clin Oncol. 1996;122(9):559-65.

70. Scholz RB, Kabisch H, Weber B, Roser K, Delling G, Winkler K. Studies of the RB1 gene and the p53 gene in human osteo-sarcomas. Pediatr Hematol Oncol. 1992 Apr-Jun;9(2):125-37.

71. Toguchida J, Ishizaki K, Sasaki MS, Ikenaga M, Sugimoto M, Kotoura Y, et al. Chromosomal reorganization for the expres-sion of recessive mutation of retinoblastoma susceptibility gene in the development of osteosarcoma. Cancer Res. 1988 Jul 15;48(14):3939-43.

72. Wadayama B, Toguchida J, Shimizu T, Ishizaki K, Sasaki MS, Kotoura Y, et al. Mutation spectrum of the retinoblastoma gene in osteosarcomas. Cancer Res. 1994 Jun 1;54(11):3042-8.

73. Horowitz JM, Park SH, Bogenmann E, Cheng JC, Yandell DW, Kaye FJ, et al. Frequent inactivation of the retinoblastoma an-ti-oncogene is restricted to a subset of human tumor cells. Proc Natl Acad Sci U S A. 1990 Apr;87(7):2775-9.

74. He J, Olson JJ, James CD. Lack of p16INK4 or retinoblastoma protein (pRb), or amplification-associated overexpression of cdk4 is observed in distinct subsets of malignant glial tumors and cell lines. Cancer Res. 1995 Nov 1;55(21):4833-6.

75. Levine AJ, Chang A, Dittmer D, Notterman DA, Silver A, Thorn K, et al. The p53 tumor suppressor gene. J Lab Clin Med. 1994 Jun;123(6):817-23.

76. Tataria M, Quarto N, Longaker MT, Sylvester KG. Absence of the p53 tumor suppressor gene promotes osteogenesis in mes-enchymal stem cells. J Pediatr Surg. 2006 Apr;41(4):624-32; discussion -32.

77. Chandar N, Billig B, McMaster J, Novak J. Inactivation of p53 gene in human and murine osteosarcoma cells. Br J Cancer. 1992 Feb;65(2):208-14.

78. Gokgoz N, Wunder JS, Mousses S, Eskandarian S, Bell RS, Andrulis IL. Comparison of p53 mutations in patients with lo-calized osteosarcoma and metastatic osteosarcoma. Cancer. 2001 Oct 15;92(8):2181-9.

79. Wang X, Kua HY, Hu Y, Guo K, Zeng Q, Wu Q, et al. p53 func-tions as a negative regulator of osteoblastogenesis, osteo-blast-dependent osteoclastogenesis, and bone remodeling. J Cell Biol. 2006 Jan 2;172(1):115-25.

80. Lengner CJ, Steinman HA, Gagnon J, Smith TW, Henderson JE, Kream BE, et al. Osteoblast differentiation and skeletal de-velopment are regulated by Mdm2-p53 signaling. J Cell Biol. 2006 Mar 13;172(6):909-21.

81. Sagartz JE, Bodley WL, Gamblin RM, Couto CG, Tierney LA, Capen CC. p53 tumor suppressor protein overexpression in os-teogenic tumors of dogs. Vet Pathol. 1996 Mar;33(2):213-21.

82. Nigg EA. Cyclin-dependent protein kinases: key regulators of the eukaryotic cell cycle. Bioessays. 1995 Jun;17(6):471-80.

83. Jiang W, Zhang YJ, Kahn SM, Hollstein MC, Santella RM, Lu SH, et al. Altered expression of the cyclin D1 and retinoblasto-ma genes in human esophageal cancer. Proc Natl Acad Sci U S A. 1993 Oct 1;90(19):9026-30.

84. Graeber TG, Osmanian C, Jacks T, Housman DE, Koch CJ, Lowe SW, et al. Hypoxia-mediated selection of cells with di-minished apoptotic potential in solid tumours. Nature. 1996 Jan 4;379(6560):88-91.

85. Kaufmann WK. Cell cycle checkpoints and DNA repair pre-serve the stability of the human genome. Cancer Metastasis Rev. 1995 Mar;14(1):31-41.

86. Lowe SW, Ruley HE, Jacks T, Housman DE. p53-dependent apoptosis modulates the cytotoxicity of anticancer agents. Cell. 1993 Sep 24;74(6):957-67.

87. Johnson AS, Couto CG, Weghorst CM. Mutation of the p53 tumor suppressor gene in spontaneously occurring osteosarco-mas of the dog. Carcinogenesis. 1998 Jan;19(1):213-7.

88. van Leeuwen IS, Cornelisse CJ, Misdorp W, Goedegebuure SA, Kirpensteijn J, Rutteman GR. P53 gene mutations in osteo-sarcomas in the dog. Cancer Lett. 1997 Jan 1;111(1-2):173-8.

89. Rodda SJ, McMahon AP. Distinct roles for Hedgehog and ca-nonical Wnt signaling in specification, differentiation and maintenance of osteoblast progenitors. Development. 2006 Aug;133(16):3231-44.

90. Dannenberg JH, Schuijff L, Dekker M, van der Valk M, te Riele H. Tissue-specific tumor suppressor activity of retinoblastoma gene homologs p107 and p130. Genes Dev. 2004 Dec 1;18(23):2952-62.

91. Clark JC, Dass CR, Choong PF. A review of clinical and molec-ular prognostic factors in osteosarcoma. J Cancer Res Clin Oncol. 2008 Mar;134(3):281-97.

92. Levine RA, Fleischli MA. Inactivation of p53 and retinoblasto-ma family pathways in canine osteosarcoma cell lines. Vet Pathol. 2000 Jan;37(1):54-61.

93. Mertens WC, Bramwell V. Osteosarcoma and other tumors of bone. Curr Opin Oncol. 1994 Jul;6(4):384-90.

94. Theilen GH, Leighton R, Pool R, Park RD. Treatment of canine osteosarcoma for limb preservation using osteotomy, adjuvant radiotherapy and chemotherapy (a case report). Vet Med Small Anim Clin. 1977 Feb;72(2):179-83.

95. Ru G, Terracini B, Glickman LT. Host related risk factors for canine osteosarcoma. Vet J. 1998 Jul;156(1):31-9.

96. Fuchs B, Pritchard DJ. Etiology of osteosarcoma. Clin Orthop Relat Res. 2002 Apr(397):40-52.

97. Wang LL. Biology of osteogenic sarcoma. Cancer J. 2005 Jul-Aug;11(4):294-305.

98. Mendoza S, Konishi T, Dernell WS, Withrow SJ, Miller CW. Status of the p53, Rb and MDM2 genes in canine osteosarco-ma. Anticancer Res. 1998 Nov-Dec;18(6A):4449-53.

99. Wang LL, Gannavarapu A, Kozinetz CA, Levy ML, Lewis RA, Chintagumpala MM, et al. Association between osteosarcoma and deleterious mutations in the RECQL4 gene in Roth-mund-Thomson syndrome. J Natl Cancer Inst. 2003 May 7;95(9):669-74.


100. Zhou H, Randall RL, Brothman AR, Maxwell T, Coffin CM, Goldsby RE. Her-2/neu expression in osteosarcoma increases risk of lung metastasis and can be associated with gene amplifi-cation. J Pediatr Hematol Oncol. 2003 Jan;25(1):27-32.

101. Flint AF, U’Ren L, Legare ME, Withrow SJ, Dernell W, Hanneman WH. Overexpression of the erbB-2 proto-oncogene in canine osteosarcoma cell lines and tumors. Vet Pathol. 2004 May;41(3):291-6.

102. Kappel CC, Velez-Yanguas MC, Hirschfeld S, Helman LJ. Human osteosarcoma cell lines are dependent on insulin-like growth factor I for in vitro growth. Cancer Res. 1994 May 15;54(10):2803-7.

103. MacEwen EG, Kutzke J, Carew J, Pastor J, Schmidt JA, Tsan R, et al. c-Met tyrosine kinase receptor expression and function in human and canine osteosarcoma cells. Clin Exp Metastasis. 2003;20(5):421-30.

104. Coltella N, Manara MC, Cerisano V, Trusolino L, Di Renzo MF, Scotlandi K, et al. Role of the MET/HGF receptor in pro-liferation and invasive behavior of osteosarcoma. Faseb J. 2003 Jun;17(9):1162-4.

105. Ladanyi M, Park CK, Lewis R, Jhanwar SC, Healey JH, Huvos AG. Sporadic amplification of the MYC gene in human osteo-sarcomas. Diagn Mol Pathol. 1993 Sep;2(3):163-7.

106. Khanna C, Wan X, Bose S, Cassaday R, Olomu O, Mendoza A, et al. The membrane-cytoskeleton linker ezrin is necessary for osteosarcoma metastasis. Nat Med. 2004 Feb;10(2):182-6.

107. Laverdiere C, Hoang BH, Yang R, Sowers R, Qin J, Meyers PA, et al. Messenger RNA expression levels of CXCR4 correlate with metastatic behavior and outcome in patients with osteosar-coma. Clin Cancer Res. 2005 Apr 1;11(7):2561-7.

108. Dickens DS, Kozielski R, Khan J, Forus A, Cripe TP. Cycloox-ygenase-2 expression in pediatric sarcomas. Pediatr Dev Pathol. 2002 Jul-Aug;5(4):356-64.

109. Priester WA, McKay FW. The occurrence of tumors in domes-tic animals. Natl Cancer Inst Monogr. 1980 Nov(54):1-210.

110. Goorin AM, Abelson HT, Frei E, 3rd. Osteosarcoma: fifteen years later. N Engl J Med. 1985 Dec 26;313(26):1637-43.

111. Withrow SJ, Powers BE, Straw RC, Wilkins RM. Comparative aspects of osteosarcoma. Dog versus man. Clin Orthop Relat Res. 1991 Sep(270):159-68.

112. Knecht CD, Priester WA. Musculoskeletal tumors in dogs. J Am Vet Med Assoc. 1978 Jan 1;172(1):72-4.

113. Gillette SM, Gillette EL, Powers BE, Withrow SJ. Radia-tion-induced osteosarcoma in dogs after external beam or intra-operative radiation therapy. Cancer Res. 1990 Jan 1;50(1):54-7.

114. Bennett D, Campbell JR, Brown P. Osteosarcoma associated with healed fractures. J Small Anim Pract. 1979 Jan;20(1):13-8.

115. Wrigley RH. Malignant versus nonmalignant bone disease. Vet Clin North Am Small Anim Pract. 2000 Mar;30(2):315-47, vi-vii.

116. Renfrew H, Rest JR, Holden AR. Extraskeletal fibroblastic os-teosarcoma in a rabbit (Oryctolagus cuniculus). J Small Anim Pract. 2001 Sep;42(9):456-8.

117. Powers BE, LaRue SM, Withrow SJ, Straw RC, Richter SL. Jamshidi needle biopsy for diagnosis of bone lesions in small animals. J Am Vet Med Assoc. 1988 Jul 15;193(2):205-10.

LABORATORY TRAINING FROM THE EXPERTS The Association of Public Health Laboratories (APHL) sponsors educational programs on critical issues in laboratory science, some of which are in conjunction with other professional laboratory organizations such as the Clinical and Laboratory Standards Institute (CLSI) and the American Society for Clinical Laboratory Science (ASCLS). Below are only a few of the courses available. All courses provide P.A.C.E. ® credits. For a complete listing of upcoming, pre-recorded, and on demand programs, visit www.laboratorytraining.org.

MARK YOUR CALENDARS (for these upcoming webinars) May 7 On Slippery Slopes: Principles, Problems and Cases in Lab Ethics May 14 The Impact of Global Warming on Vector-Borne Diseases June 4 Determining Clinical Relevance of Fungal Isolates June 20 Transport Systems and CLSI Document M40: It’s Not Just Swabs (cosponsor CLSI) June 27 CLSI’s Guide to Planning Laboratory Operations during a Disaster (cosponsor CLSI) Above are just a few of the upcoming webinars. Registration fee for webinars includes the live webinar for ALL of your staff PLUS unlimited access to the archived webinar (and CEUs) for your staff for 6 months! What a bargain! Don’t worry if you can’t participate on that day; all are available as on-demand courses AFTER the course—same great deal of unlimited access (and CEUs). AND MANY MORE! Log on to www.laboratorytraining.org and see why thousands of your colleagues attend APHL-sponsored courses each year!

REGISTRATION NOW OPEN: 2013 APHL Annual Meeting & 7th Government Environmental Laboratory Conference June 2-5, 2013 Raleigh, NC www.aphl.org/2013annualmeeting Pre-Conference Workshops (June 2): “FBI’s Joint Criminal & Epidemiological Investigation” & “Informatics & Meaningful Use” www.aphl.org/2013annualmeeting

www.aphl.org


1. Treatment for Osteosarcoma has not changed in the last 30 years.

A. True B. False

2. Why have researchers selected the tibial injected model of OS in mouse?

A. Because of the similarity in oncogenic behaviors between human & mouse model

B. Because of the similarity in clinical diagnosis between human & mouse models

C. Because the similarity in tumorogenesis & metastasis behavior between human & mouse model

D. Because the similarity in environmental factors between human & mouse models

3. Bioluminescence, an in vivo imaging technique, enables real-time monitoring of the labeled substances in experimental animals.

A. True B. False

4. Cure rates for patients with metastatic or relapsed disease are poor, around 50% survival.

A. True B. False

5. Osteosarcoma metastases (tumor invasiveness) by blood vessels

A. True B. False

6. Osteosarcoma has high invasiveness and metastasis into the

A. brain and CNS B. lower limbs C. lung and liver D. adrenal gland & kidney

7. The cellular similarity between double conditional knockout mouse model and tibial injected mouse model of Osteosarcoma is:

A. Both models of Osteosarcoma cells are difficult to culture in vivo

B. Both cells are mechanically disaggregated C. Both cells show the invasiveness to the

kidney tissue in the immune-compromizing node mice

D. Both cells are highly differentiated

8. The genetic engineering techniques had been performed on osteoblast - knockout of p53 and pRb using Osx1 - Cre, which is called double conditional knockout (DKO). The transgenic model has early developed OS with

A. High penetrance and short latency. B. High penetrance and long latency C. Low penetrance and short latency D. Low penetrance and long latency

9. In bone malignancies, the loss of pRb can suppress the terminal stage of:

A. Osteogenic differentiation B. Chondogenic differentiation C. Embryonic Stem cell differentiation D. Hematopoietic cells differentiation

10. p53 gene has unrecognized ability to modulate the differentiation of osteoblasts, and bone development.

A. True B. False




Article 400 2 Clock Hours


HOME STUDY UNITS FOR AMT MEMBERSOffered by Association for Continuing Education, LLC, (ACE)

The self-instructional units listed below have been reviewed and approved for Continuing Education for AMTmembers by the American Medical Technologists Institute for Education (AMTIE).

To participate in home study programs: 1) Order the units desired directly from Association for ContinuingEducation (ACE) using the form below (photocopy accepted); 2) Complete unit and AMTIE post test enclosedwith unit; 3) Send completed post test to ACE, P.O. Box 573, Beaufort, SC 29902 with $6.00 per test for gradingand score reporting. Results of your participation will be recorded in your AMT continuing education file.

ORDER FORMThe following self-instructional units are AMTIE approved for AMT Continuing Education.

Total ___________

Shipping ___________

Amount Enclosed ___________

Return this form with check or money order to:ACEP.O. Box 573, Beaufort, SC [email protected]

PROGRAM CLOCK UNIT QUANTITY TOTALHOURS COST ORDERED COST

Basic Laboratory Techniques#100 Performing a Capillary Puncture 1.5 $ 9.50 _________ _________#101 Venipuncture: The Art of Drawing Blood 3.0 $13.50 _________ _________#102 The Making of a Blood Film 1.5 $ 9.50 _________ _________

Chemistry#206 Quality Control Overview for Clinical Chemistry 3.0 $13.50 _________ _________#207 Laboratory Evaluation of Cardiac Markers 3.0 $13.50 _________ _________#208 Kidney Function Tests 3.0 $13.50 _________ _________#209 Total and Ionized Calcium in Serum 2.0 $11.50 _________ _________

Chemistry#400-1 Intro to Hematopoiesis - booklet and CD w/35 photo images 4.0 $37.50 _________ _________#400-2 Intro to Hematopoiesis - booklet only (no CD) 2.0 $12.50 _________ _________#404 Hematology Indices 2.0 $12.50 _________ _________#409-1 Cerebrospinal Fluid - booklet and CD w/37 photo images 5.0 $44.50 _________ _________#409-2 Cerebrospinal Fluid - booklet only (no CD) 3.0 $13.50 _________ _________#410-1 Reticulocyte Counts - booklet and CD w/35 photo images 4.0 $37.50 _________ _________#410-2 Reticulocyte Counts - booklet only (no CD) 2.0 $12.50 _________ _________#411 Erythrocyte Sedimentation Rates 1.0 $ 8.50 _________ _________#414 Hemoglobin H Disease 2.0 $12.50 _________ _________#415 Iron Metabolism 3.0 $13.50 _________ _________#418 Hemolysis Testing 4.0 $14.50 _________ _________#450 Coagulation Phase of Hemostasis 2.0 $12.50 _________ _________

Immunology#500 Intro to ABO Blood Group System 2.0 $12.50 _________ _________#501 Reading and Grading Agglutination Reactions 1.0 $ 8.50 _________ _________#502 Solving Blood Bank Problems 3.0 $14.50 _________ _________#503 Problems in Antibody Identification 3.0 $14.50 _________ _________#550 Antigens and Antibodies 2.0 $12.50 _________ _________#551 Principles of Antigen-Antibody Reactions Used in the Lab 3.0 $14.50 _________ _________#552 Complement Cascade 2.0 $12.50 _________ _________

Microbology#606 Overview of TB Infection and Disease 2.0 $12.50 _________ _________

Mycology#650 Introduction to Medical Mycology 1.0 $ 8.50 _________ _________

Statistics#762 Descriptive Statistics 2.0 $12.50 _________ _________

Urinalysis#800 Chemical Screening of Urine by Reagent Strip 2.5 $13.00 _________ _________

Name ______________________________________________________

Address ____________________________________________________

City, State, Zip_______________________________________________

___MT ___MLT ___RMA ___RDA

AMT ID #_________________________

$4.75

Continuing Education OpportunitiesAMT strongly advocates lifelong learning as a part of your personal and professionalgrowth. Take a look at the great CE opportunities that AMT has to offer!

STEP Online

STEP Online consists of short scholarly articles on a variety of topics and a 10-question quiz. While the articles and quizzes are available

online, they also appear in the Journal of Continuing Education Topics & Issues. A nominal fee is assessed for the quiz, taken either

online or manually.

Topics

On-Demand Courses

On-demand courses are offered on timely topics and are available 24/7. Each course consists of an audio-presentation synched with

PowerPoint slides. To receive credit, the participant must successfully pass a 10-question quiz. A nominal member fee applies.

Webinars

available 24/7. To receive credit, the participant must successfully pass a 10-question quiz. A nominal member fee applies.

Topics

Prevention

Assistants


When looking to learn about proper chemical storage in your laboratory, consult officials who can advise you. These might include your facili-ty’s safety officer, the local fire marshal, a radia-tion safety officer (if you handle radioactive ma-terial), an Occupational Safety and Health Administration representative and/or an Envi-ronmental Protection Agency official (if you’re disposing of unused chemicals). “Become friends with these people, they can help you,” ad-vised Peggy A. Wenk, HTL(ASCP)SLS, Beau-mont Health System, Beaumont Laboratory, Royal Oak, MI, who recently presented an audio conference on this topic for the National Society for Histotechnology.

When consulting with these individuals, tell them what chemicals you have, where they are located, the room’s layout, where fire extinguish-ers are placed and where chemicals are being stored. “You don’t want safety personnel to be surprised during a fire or evacuation,” Wenk said. “Invite them to come in and to look around. Ask them for advice and to train you on how you can do things better.”

“They can’t cite or fine you if you invited them in,” Wenk continued. “That is much better than them coming in unannounced and finding viola-tions and fining you. That can be very expen-sive.”

Preparing a Chemical InventoryCreate an inventory sheet of all chemicals in

stock prior to a consultant’s visit. Indicate details such as the quantity on hand and its physical state (e.g., solid, liquid or gas). This will save a significant amount of time and should be easy to revise. New employees can access this form to determine what type of gloves to wear, how to

store chemicals, how to dispose of chemicals and so forth.

If you need a safety data sheet, ask your safety officer or google “MSDS” and the chemical’s full name in quotes. Enter vendor information and any other relevant information such as correct chemical, correct percent and concentration. You can also obtain a lot of MSDS’ from the web site “Safety Information Resources, Inc.”, avail-able at http://siri.org/msds/index.php.

On the chemical inventory sheet, be sure to re-cord the chemical’s exact name. This might re-quire writing more than one name, such as sodi-um borate, borax or sodium tetraborate decahydrate.

If you’re listing dye powders, include the digi-tal number from the Biological Stain Commis-sion. “If using pre-made solution, you don’t have to list every single dye,” Wenk noted.

Your inventory sheet should also include where each chemical is stored, how to dispose of it, if any personal protective equipment needs to be worn when handling it, what engineering controls need to be used, if a splash guard is re-quired, if it needs to be used in a hood, and so forth.

Properly Placing ContainersChemicals should be stored at or below coun-

tertop level. They should not be stored on the floor (they can be kicked over and break) or a top shelf (they could fall and break). Don’t store too many chemicals in a cupboard. Having to move around containers to find the correct chemical could lead to a container being dropped.

Don’t store chemicals underneath a sink, ei-ther. Hot water passing through pipes can warm

Karen Appold is a medical writer. Visit www.WriteNowServices.com.

Safely Storing Laboratory Chemicals

Karen Appold



an area, heat up chemicals and cause them to break down. Also be wary of storing too many bot-tles in a hood--this can impair ventilation.

Chemicals also shouldn’t be stored on the coun-tertop. Here, they can be knocked over and ex-posed to light, exposing employees to additional dangers. Wenk advised against storing chemicals alphabetically, because some chemicals aren’t compatible. “Store chemicals according to their hazard,” she said.

You’ll also want to avoid placing chemicals in sunlight or direct light. “Bulbs that illuminate counters make the counters hotter, lighter and brighter,” Wenk said. “Pay attention to light and heat sources. Put chemicals in cupboards away from heat, including the undercounter lights that warm up the bottom shelf in the cupboard.”

Wenk also suggested that laboratorians examine shelves and racks. Make sure they are strong enough to support the weight of chemicals. Don’t store bottles on their sides if they don’t fit upright in a cupboard--they could roll off and break. Don’t store bottles on top of each other, either. Securely anchor shelves, cupboards and racks to a perma-nent structure. Put a bar or lip on the front of a shelf in case of an earthquake or other unsuspect-ing motion occurs. “And don’t think that some-thing couldn’t happen to you,” Wenk said. “People in Washington, DC, were totally taken off guard with the earthquake in 2011.”

Clean out cupboards periodically. “If you don’t use a chemical for one year, throw it out. Same goes if the expiration date occurs, a container rusts or bulges, or the color changes,” Wenk said. “Purchasing large quantities isn’t always wise if you have to toss unused expired chemicals.”

Choosing ContainersAs far as the type of material used to store chem-

icals, plastic is ideal. Metal containers can rust and/or the label might fall off. Spills and splashes could result. Glass can break. But if glass is neces-sary, Wenk advised buying glass coated with poly-ethylene that is impact resistant. Always check for leaks and spills when storing chemicals.

Ideally, chemicals should be stored in their orig-inal containers. “You can pour a chemical into an-other container, but be sure to properly label it,” Wenk said. Check that lids and caps are on tightly. “Chemicals can outgas and they can interact and create new chemicals,” Wenk noted.

Signs and LabelsSome dangers can be avoided by properly iden-

tifying hazardous chemicals and areas. For in-stance, make large signs to indicate the oxidizer cabinet and note where the flammable cabinet is. Post a note on a refrigerator if food shouldn’t be stored in it. Many companies sell safety signs, one of which is “Lab Safety Supply,” available at www.lss.com.

Label chemicals with the dates they arrived and when they expire. “If you make chemicals, you need to say exactly what chemicals are in the solu-tion,” Wenk reminded listeners.

Specific Chemical Storage TipsWenk also provided specific advice for the stor-

age of all types of chemicals, from acids to reac-tive/explosive chemicals. Here’s a partial summa-ry of tips she offered.

• Store acids in a corrosive cabinet that has the word “acid” written across it. Certain acids should not be stored together. For example, nitric acid should be stored separately from organic and inorganic acids.

• Bases/alkalis, the opposite of acids, need to be stored separately in a corrosive-resistant cabinet. They can destroy tissue and/or met-als.

• An oxidizer, a chemical that releases oxygen, will increase a fire hazard. Don’t store them near paper.

• Contact your safety department and/or fire marshal to figure out where flammables/com-bustibles can be stored, and at what quantity.

• Reactive/explosive chemicals, i.e., those that will have a sudden release of pressure, gas or heat, should be stored in places where mini-mal damage would occur.

• Toxic chemicals, those that will act upon life processes, and cause death, temporary inca-pacitation, or permanent harm to humans or animals, should be stored in a cool, well-ven-tilated area, away from heat and light.

• Controlled substances, i.e., drugs or other substances that have a potential for abuse or for causing dependence, should be in a dou-ble locked metal box, bolted to a permanent structure.


Crossword solution from page 42

Final ThoughtsWenk advised laboratorians to walk around

their lab with “chemical eyes looking for chemical risks.” Consult safety experts to determine the risk assessment, create a chemical inventory sheet, im-plement appropriate storage techniques, and iden-tify hazards and plan to minimize them.

This article is based upon an audio conference presented by the National Society for Histotech-nology titled, "Safe Storage of Laboratory Chemi-cals,” given by Peggy A. Wenk, HTL(ASCP)SLS, Beaumont Health System, Beaumont Laborato-ry, Royal Oak, MI.


1. Which personnel can provide information on how to properly store chemicals in your laboratory?

A. Your facility's safety officer B. The local fire marshal C. A radiation safety officer D. All of the above

2. If you invite safety personnel into your lab, they can cite or fine you for a violation.

A. True B. False

3. Chemicals can be stored anywhere in the lab, as long as they are stored properly.

A. True B. False

4. You shouldn't store chemicals underneath a sink because hot water going through pipes can heat up chemicals and cause them to break down.

A. True B. False

5. It's a good idea to store chemicals alphabetically, because they will be easier to find.

A. True B. False

6. Which one is not a good reason to throw away a chemical:

A. It just expired. B. You haven't used it for six months. C. The container is bulging. D. A chemical's color changed.

7. What is the best type of material to store most chemicals in?

A. Metal B. Glass C. Plastic D. Cardboard

8. Certain types of acids should never be stored together.

A. True B. False

9. Which type of chemical should not be stored close to paper?

A. Acid B. Oxidizer C. Bases D. Alkalis

10. What are some guidelines regarding storing toxic chemicals?

A. Store in a cool place B. Store in a well-ventilated area C. Store in a well-lit area D. Only A and B are correct.






AMT, established in 1939, is a national, non-profit certification agency for:

Medical Technologist, MT®

Medical Laboratory Technician, MLT® Registered Medical Assistant, RMARegistered Dental Assistant, RDACertified Medical Laboratory Assistant, CMLARegistered Phlebotomy Technician, RPT Certified Laboratory Consultant, CLCCertified Allied Health Instructor, CAHICertified Medical Administrative Specialist, CMASFor information on qualifications necessary for

each certification, contact: AMT, 10700 Higgins Rd., Suite 150, Rosemont, IL 60018—Phone: 847/823-5169.

AMT OFFICEChristopher A. Damon, J.D., Executive Director

10700 W. Higgins Road, Suite 150 Rosemont, Illinois 60018

Telephone: (847) 823-5169 or (800) 275-1268Fax: (847) 823-0458

E-mail: [email protected] Site: http://www.americanmedtech.org

AmericAn medicAl TechnologisTs AMT NATioNAl officers, 2013 (Terms of office)

President: Mary Burden, MT, 1041 Kings Road, Moore, OK 73160 (2009–2015)Vice-President: Everett Bloodworth, MT, 930 Pine St., Benton, KY 42025 (2011–2014)secretary: Jeff Lavender, MT, 13990 Blue Haven Dr. NE, Kalkaska, MI 49646-9150 (2009-2015)treasurer: Janet Sesser, RMA, 2815 East Windrose Dr., Phoenix, AZ 85032-6554 (2011–2014)

BoArd of direcTors (Terms of office)

Nancy B. Barrow, MT, 715 Beverly Way, Martinsville, VA 24112 (2010–2013)Dr. Paul C. Brown, MT, 2506 South Cobb Loop, Millbrook, AL 36054 (2010–2013)

Heather Herring, MT, RMA, 20404 E. Eaves Way, Farmington, MN 55024 (2012-2015)Jeannie Hobson, RMA, RPT, CMAS, AHI, 2323 E. Robinson, Fresno, CA 93726, (2011-2013)Carole Aston, 716 Ontario St. #3, Oak Park, IL 60302 (Public Member) (Appointed annually)

iMMediATe PAsT-PresideNTRoxann Clifton, MT, 409 E. Mississipi, Sayre, OK 73662 (2011–2014)

JudiciAry couNcillor (Appointed annually)Kim Cheuvront, PhD, 100 Fair Oaks Dr., Fairmont, WV 26554

eXecuTiVe couNcillor (Appointed annually)Edna Anderson, MT, 1397 Redwood St., NW, Salem, OR 97304

disTricT couNcillors (Appointed annually)

AMTie BoArd of TrusTees, 2013 (Terms of office)President: Linda Jones, MT, 4673 Lambsburg Rd., Lambsburg, VA 24351 (2012-2015)Vice-President: Art Contino, RMA, 3117 South Horizon Pl., Oviedo, FL 32765 (2011-2014)secretary Marty Hinkel, MT, 2204 Peggy Dr., Worland, WY 82401 (2010-2013)treasurer: David P. Yocom, Jr., 4121 119th St. SE, Everett, WA 98208-5344 (Public member - appointed annually)executiVe director: Gerard P. Boe, PhD, MT, 7 Sussex Ct., Beaufort, SC 29907 (Appointed annually)trustees: Everett Bloodworth, MT, 930 Pine St., Benton, KY 42025 (AMT Board appointment)(Appointed annually) Norma (Taffy) Durfee, MT, PO Box 432, Iola, TX 77861 (2012-2015) Kay Fergason, MT, 3712 Arava Dr., Green Cove Springs, FL 32043 (Immediate Past President) (2009-2013) Zenaida Maraggun, MT, 1602 Amour Drive, Leesville, LA 71446-5215 (2011-2014)

eastern district(Maine, New Hampshire, Vermont,

Massachusetts, New York, Connecticut, Rhode Island, New Jersey, Pennsylvania, Delaware, District of Columbia, Maryland,

West Virginia, Canada)Janet Crigler, MT, 23 Pheasant Dr.,

Fairmont, WV 26554

Great Lakes district (Michigan, Wisconsin, Illinois, Indiana, Ohio, Iowa, Minnesota, North Dakota,

South Dakota)Clara Boykin, MT

1023 Dayton Ave., St. Paul, MN 55104

southern district (Alabama, Florida, Georgia, South Carolina,

Kentucky, North Carolina, Tennessee, Virginia, Caribbean)

Shannon Newman, MT 249 Willie Craig Rd. Bassett, VA 24055

Western district(Washington, Oregon, Idaho, Montana, Ne-

vada, California, Wyoming, Utah, Colorado, Arizona, New Mexico,

Alaska, Hawaii) Kenneth Hawker, MT

6631 W. Sunrise Oak Dr.West Jordon, UT 84081

central district(Texas, Oklahoma, Arkansas, Louisiana, Nebraska, Kansas, Missouri, Mississippi)

Randall Swopes, MT2691 Whittington

Westlake, LA 70669


direcTory

ExecutiveMary Burden, MT, PresidentEverett Bloodworth, MT, Vice

PresidentJeffrey Lavender, MT, SecretaryJan Sesser, RMA, TreasurerRoxann Clifton, MT, Immediate Past

PresidentEdna Anderson, Executive

CouncillorKim Cheuvront, PhD, Judiciary

CouncillorChristopher A. Damon, J.D.,

Executive Director (ex-officio)

Audit & BudgetJanet Sesser, RMA, ChairEverett Bloodworth, MTMary Burden, MTLuther Ray Dean, MTCynthia Kukenberger, MTMiriam “Mia” Rutkiewicz, RMANicole Weiss-Lopez, RMA, RPTChristopher A. Damon, J.D.,

Executive Director (ex-officio)

Federal Government/LegislativeJohn Sherer, MT, ChairPat Westbrook, MT, Vice ChairMary Burden, MTGeorge Cook, MTBill Dettwyler, MTLinda Jones, MTMary Midkiff, MTBob Newberry, MTBarbara Ware, MTKimberly Cheuvront, PhD (ex-officio)Michael McCarty (ex-officio)

JudiciaryKim Cheuvront, PhD, ChairEverett Bloodworth, MTSheryl Rounsivill, RMA, RPT, CMAS,

AHIMichael McCarty, Legal Counsel

BylawsKimberly Cheuvront, PhD, ChairJoyce Lybrand, MTLinda Raven, RMA, RPT, COLTMichael McCarty (ex-officio)

Examinations, Qualifications and StandardsDavid McCullough, MT, ChairRoxann Clifton, MTRonald Lepoff, MDJanet Sesser, RMAJames Fidler, PhD (ex-officio)

Laboratory Certification Examination Development TeamsGeneral LaboratoryCharles Baker, MT, Team LeaderFred Morley, MTNorma (Taffy) Durfee, MT Clinical ChemistryMary Midkiff, MT, Team LeaderRoxann Clifton, MT

Hematology, Hemostasis, and UrinalysisDr. Georgia McCauley, Team LeaderCarlo Ledesma, MTYvonne Spade, MT Blood Banking, Immunohematology, and ImmunologyJudy Smith, MT, Team LeaderMichele Gillies, MTCynthia Kukenberger, MT MicrobiologyDr. Terry Else, Team LeaderDr. Joel MortensenKathy Sutton, MT Certified Medical Laboratory Assistant (CMLA) Exam TeamBarbara Ware, CLC, ChairAnn BachmanChris Pontious, AHI, COLT, RMA, RPTBob Newberry, MTLinda Jones, MTJames Fidler, Ph.D. (ex-officio)

RMA EQSClinical TeamDeborah Westervelt, COLT, RMA,

Committee Co-Chair, Clinical Team Leader

Arthur Contino, AHI, RMAHeather Herring, MT, RMAJeannie Hobson, RMA, RPT, CMAS,

AHITracy Mixdorf, D.O.James Fidler, PhD (ex-officio) Administrative TeamJill Carlson, RMA, Committee

Co-Chair, Administrative Team Leader

Beverly Christiansen, RMAMarilyn Johnson-Gilliam, RMA, RPTDonna Nelson, RMAPatricia Poitier-Sands, RMAAngela Davis-Woodson, RMAJames Fidler, PhD (ex-officio)

RPT EQSRichard Crowner, MT, RPT, ChairMarty Hinkel, MTAlice Macomber, RMA, AHI, RPTLinda Raven, RMA, RPT, COLTDorothy Roush, MT (Emeritus)Anna Seals, RPTJames Fidler, PhD (ex-officio)

RDA EQSJudith Dry, RDA, ChairVivian KoistinenGlinda Otoki-GarrettLisa TheodoreAlthea WynnJames Fidler, PhD (ex-officio)

CMAS EQS Sharon Paff, RMA, ChairBarbara Garrido, RMAKathleene Hardy, RMADonna Hiatt, RMA, CMASDiana Kendrick, RMAJames Fidler, PhD (ex-officio)

State LegislativeBob Newberry, MT, ChairEverett Bloodworth, MTGeorge Cook, MTAll State Legislative ChairsTerri Breitwieser, MT, AHICarletha Durham, MTWebb Gray, MTLinda Raven, RMA, RPT, COLTDiane Robbins, MTJohn Sherer, MTBarbara Ware, MTPat Westbrook, MTDeborah Westervelt, RMA, COLTTommie Williams, MTMichael McCarty (ex-officio)Kimberly Cheuvront, PhD (ex-officio)

CASMET LiaisonChris Seay, MT

CredentialsJeff Lavender, MT, ChairJeannie Hobson, RMA, RPT, CMAS,

AHIJanet Crigler, MTCecil Hunt, MTMarilyn Johnson-Gilliam, RMAChristopher Seay, MT

Future Planning/MembershipMary Midkiff, MT, ChairTaffy Durfee, MT, Co-ChairEdna Anderson, MTNancy Barrow, MTAlicia Gregorio, MTJosephine Harden, MTDebra Janeczko, RMAJeffrey Lavender, MTLucy Leyva, RPTBrett Merkle, RMAFrancine Oran, RMAAnn Roby, COLTLinda Witte, RMACarol Yankovich, MTChristopher Damon, JD (ex-officio)Kathy Cilia, MT (ex-officio)

ProctoringEverett Bloodworth, MT, ChairAll State Society Proctor ChairsKathy Cilia (ex-officio)

NominatingLouise Isbell, RMAMelissa Martin, RMAKim Meshell, RMA, RPT, AHISueollen Schobert, MLTDave McCullouth, MT

NominatingAlternatesCharles Baker, MTLeann Bart, RMAVernell Boyd, MTJohn Sherer, MTBarbara Ware, MT

ConventionPeggy Oiler, MT, ChairJanet Crigler, MTGloria Culla, MTLynn Dunlop, MLTCarla Filles, RPTHeather Herring, MT, RMAMarty Hinkel, MTAlicia Houston, MLTJerry Hudgins, MTElizabeth Sarchet, MLTLia Kaye Spears, MTDiane Powell, CMP (ex-officio)

Scientific/SpeakersKay Fergason, MT, ChairMichelle Jenkins, MT, Vice ChairJeri Bond, RMA, AHIDon Bouchelle, MTClara Boykin, MTTereyo Cop, MTSujana De Almeida, RMAArlene DeCarli, MTRoxanne Erskine, MTHattie Gallon, MTLt. Calvina Glover, MTSolomon Goldberg, RMAPatty Harris, MTVern Hein, MTPam Kriegel, MTDorothy Marks, MTMelissa Martin, RMANaomi Melvin, MTFred Morley, MTJean Palmer, RMA, AHIPatricia Poitier-Sands, RMADelores “Lola” Rosalis, RMAMimi Roush, MTAlberta Smith, RMA, AHISieglinde Wildie, MLTFelicia Williams, MTDiane Powell, CMP (ex-officio)

MentorBarbara Ware, MT, ChairTera Benefiel, MTNathalie Dixon, MTBetty Geary, MTAlice Macomber, AHI, RPTJuanita Stocke, MTAudrienne Whitley, MTVirgil Marchand, RMA Student ActivitiesJulie Hardcastle, MT, ChairIvette Rivera, RMA, AHI, Vice ChairVanessa Austin, AHI, RMALisa Marie Bromley, RMAJamie Horn, RMA, RPT

Elizabeth Hurd, MTAmber Huskinson, MLTKarina Ibarra, RMA, RPTAlaine Johnson, RMA, AHICynthia Jones, MTKody Karas, RPTGrant Lambert, MTFred Morley, MTShannon Newman, MTRikki Packer, RMAChris Pontious, RMAFaith Robeson, MA StudentOzzie Skinner, MTLeonila Sumarsono, RMA, RPTLynette Thomson, MTNicole Weiss, RMA, RPTKathy Cilia, MT (ex-officio)

PublicationsNancy Gabi, RMA, ChairMaria Chevy-Newham, MTCarole Fecteau, MTMaria Guzman, MTKen Hawker, MTTeresita Hacuman, AHIKim Meshell, RMA, RPT, AHIRobin Miliner, MTDonna Nelson, RMASuellen Schobert, MLTEdith Tefft, MTKaye Tschop, MTDiane Powell, CMP (ex-officio)

Armed ServicesCalvina Jordan, MT, ChairSarah Hanaway, MLTGerald Simi, MLT, AHIRandy Swopes, MTFelicia Williams, MTViviana Vera, RMAChristopher Damon, JD (ex-officio)

CLC Evaluation CommitteeGerard P. Boe, PhD, ChairAnn Bachman, CLCJoel E. Mortensen, PhDAnn M. Steele, PhDDianne B. Zielinski, PhDChristopher A. Damon, J.D.

(ex-officio)

Career Education Advisory CommitteeBradley Moore, ChairJanice DonnellyJudith Dry, RDASandy OckTammy RennerMarylou de-Roma-RagazaJanet Sesser, RMAJohn Smith, Ed.D.Christopher A. Damon, J.D.

(ex-officio)Kathy Cilia (ex-officio)

2013 Committees

ALABAMA Rikki Packer, RMA, Virginia Col, 17634 Dailey Lane, Foley, AL 36535, email: [email protected] Fred Morley, MT, 5700 East El Camino Quinto, Apache Junction, AZ 85119, email: [email protected] Tonda Ellis, CMLA, RPT, 1160 Midway Rd., Monticello, AR 71655, email: [email protected] Sheryl Rounsivill, RMA, RPT, CMAS, AHI, 2078 S. Hayston, Fresno, CA 93702, email: [email protected] PLAInS (kansas/nebraska) Tera Benefiel, MT, 622 S. McPherson Ave., Burrton, KS 67020, email: [email protected] – (See Rocky Mountain)COnnECTICUT – (See Tri-State)DC/DELAWARE/MARYLAnD Robin Miliner, MT, 9695 Halstead Ave., Laurel, MD 20723, email: [email protected] Kay Fergason, MT, 3712 Arava Dr., Green Cove Springs, FL 32043, e-mail: [email protected] Marvin Matthews, MT, 5565 La Fleur Trail, Lithonia, GA 30038, email: [email protected] Minelva B. Manuel, RMA, 98-410 Koauka LP Unit 10F, Aiea, HI 96701 e-mail: [email protected] (See Northwest)ILLInOIS Nancy Gabl, RMA, AHI, 1768 Coach Drive, Naperville, IL 60565, email: [email protected] L.E. Vern Hein, MT, 6060 E. 141st Ave., Crown Point IN 46307-9254IOWA Beverly Christiansen, RMA, 1096 Grouse Ave., Hampton, IA 50441, email: [email protected] – (See Central Plains)kEnTUCkY Christina Huff, RPT, RMA, 597 Morehead Rd., Bowling Green, KY 42101, email: huffcggclinic.comLOUISIAnA Zenaida Maraggun, MT, 1602 Amour Drive, Leesville, LA 71446, email: [email protected]

MAInE/nEW HAMPSHIRE Susan Constable, MT, 257 Heywood Rd., Winslow, ME 04901, email: [email protected] – (See Tri-State)MARYLAnD (See DC/Delaware)MICHIGAn Sieglinde Wildie, MLT, 25 Rural St., Port Huron, MI 48060MInnESOTA Edith Tefft, MT, 317 Frenn Ave., Red Wing, MN 55066, email: [email protected] Cecil Hunt, MT, 4040 Colton Dr., Olive Branch, MS 38654, email: [email protected] Alberta Smith, RMA, AHI, 1997 Ridgeway, Arnold, MO 63010, email: [email protected] (See Northwest)nEBRASkA – (See Central Plains)nEVADA Juanita Stocke, MT, 1812 Cambridge Hills Ct., Reno, NV 89523, email: [email protected] HAMPSHIRE – (See Maine)nEW jERSEY Elizabeth Suarez, RMA, Lincoln Tech Inst, 3 Silvercolt Dr., Colts Neck, NJ 07722, email: [email protected] MExICO Virgil E. Marchand, RMA, Pima Med Inst, 3501 Santa Teresa NW, Albuquerque, NM 87120-3627, email: [email protected] YORk Camille McIntyre, MT, 4401 Matilda Ave., Bronx, NY 10470, email: [email protected] CAROLInA Jerry Johnson, MT, 1296 Reeves Mill Rd., Mt. Airy, NC 27030, email: [email protected] (Washington State/Idaho/Montana) Jo Abraham, RMA, 25032 SE 384th St., Enumclaw, WA 98022, email: [email protected] Christopher Williams, RMA,AHI, 6637 Hubbard Dr., Huber Heights, OH 45424-3534, email: [email protected]

OkLAHOMA Kimberly Digby, MLT, 2013 Brighton Ave., Oklahoma City, OK 73120, email: [email protected] Marilyn Albertsen, MT, 88515 Hwy. 202, Astoria, OR 97103, email: [email protected] John A. Rudnick, MT, 501 Locust St., Greensburg, PA 15601RHODE ISLAnD – (See Tri-State)ROCkY MOUnTAIn (Colorado, Wyoming) Jennifer Dillard, RMA, 16324 E. 107th Pl., Commerce City, CO 80022, email: [email protected] CAROLInA Peggy McCutcheon, MT, 941 McCutchen Rd., Cades, SC 29518, email: [email protected] Jerry T. Hudgins, MT, 221 Windsor Park Ln., Hendersonville, TN 37075, email: [email protected] Taffy K. Durfee, MT, PO Box 432, Iola, TX 77861, email: [email protected] (Connecticut, Massachusetts, Rhode Island) Phyllis Nordby, RMA, Porter & Chester Inst., 166 Davis St., Oakville, CT 06779, Email: [email protected] Michelle Tew, RMA, 1158 Lafayette Drive, Salt Lake City, UT 84116, email: [email protected] Patricia Harris, MT, P.O. Box 262, Independence, VA 24348WASHInGTOn STATE – (See Northwest)WEST VIRGInIA Tonya Brown, MT, 333 Baldwin St., Fairmont, WV 26554, email: [email protected] Julie Lent, MT, 610 Fremont St., Algoma, WI 54201, email: [email protected] – (See Rocky Mountain)CARIBBEAn ASSn. (CASMET) Grant Lambert, AHI, PO Box 2293, Lagoon Rd., St. George’s, Grenada, West Indies, email: [email protected]

72 April 2013 • Continuing Education Topics & Issues72 January 2013 • Continuing Education Topics & Issues

ABSTRACTS FROM THE CURRENT LITERATURE

None of us can read all the medical literature, even that that pertains particularly to medical technology. Presented below are short abstracts from current literature presented with the hope that they will answer some of your questions and lead you to a better understanding of what is happening. You are encouraged to send copies of articles you have found in journals or on the Internet to AMT and we will abstract them for an upcoming issue. We encourage and welcome future contributors from readers of this journal. Please send your abstracts to Editor, Journal of Continuing Education Topics & Issues, 10700 W. Higgins Rd., Suite 150, Rosemont, IL 60018.

The following abstracts were contributed by David Plaut, Plano, TX, who is AMT’s book reviewer and a frequent speaker at AMT annual conventions.

Your hospital librarian or your public librarian can help obtain copies of the full text of these articles.

Iron deficiency and overload. Implications in oxidative stress and cardiovascular health. Nutr Hosp. 2010 May-Jun;25(3): 350-65. Toxqui L. De Piero A. Courtois V. et al.

Although iron is an essential mineral for maintaining good health, excessive amounts are toxic. Nowadays, much interest is focused on the mechanisms and regtulation of iron metabolism by downregulation of the hor-mone hepcidin. Disorders of iron metabolism could lead to iron overload, mainly causing the rare disease he-reditary hemochromatosis, or on the other hand, iron deficiency may cause anemia. Currently, these alter-ations constitute an important problem of public health. Iron overload causes lipid peroxidation and increases cardiovascular risk. Recently, a relationship between iron metabolism and insulin resistance and obesity has been described. In contrast, regarding a possible relationship between iron deficiency anemia and cardio-vascular disease, many aspects remain controversial.

Clustering and risk factors of methicillin-resistant Staphylococcus aureus carriage in two Italian long-term care facilities. Infection. 2009 Jun;37(3): 216-21.Brugnaro P, Fedeli U, Pellizzer G

Methicillin-resistant Staphylococcus aureus (MRSA) is a well-recognized agent of health care-associated infections in long-term care facilities, but few data about the circulation of MRSA in this setting in Italy are available. The aim of the study is to determine the prevalence and risk factors for MRSA carriage in nursing home residents in Vicenza (northeastern Italy). A point prevalence survey was conducted in two long-term care facilities (subdivided into 15 wards) from 12 June 2006 to 6 July 2006. Anterior nasal swabs were ob-tained from residents and laboratory screening for MRSA was performed; full antibiotic susceptibility was as-sessed in MRSA isolates. Macrorestriction analysis of chromosomal DNA was carried out by pulsed field gel electrophoresis (PFGE). For each subject, demographic data, length of stay, dependency, cognitive function, presence of medical devices, comorbidities, current and previous antibiotic treatment, previous hospital ad-mission and presence of infection were assessed on the day of sample collection. Nasal swabs were ob-tained in 551 subjects; overall 43 MRSA carriers were detected. The rate of nasal carriers was very similar in the two institutions, and varied from 0% (0/36) to 18% (7/39) between wards. Only two out of 15 wards were found to have no MRSA carriers; overall, three pairs of colonized roommates were detected. Upon multilevel logistic regression, the risk of MRSA carriage was increased in patients with cancer in those that had under-gone recent hospitalization and it reached an outcome ratio of 4.0 in those with three or more antibiotic treat-ments in the previous year; about 10% of the variability in MRSA carriage could be attributed to differences between wards. Pulsed field gel electrophoresis analysis permitted the definition of six clusters; two of these comprised 78.6% of the studied isolates and were quite similar, with one being more strongly represented among subjects hospitalized in the previous 12 months. All of the MRSA strains were resistant to ciprofloxa-cine; nevertheless, the majority were susceptible to most other non-betalactam antibiotics. The study sug-gests that nursing homes are a significant reservoir for MRSA. Statistical and PFGE analyses indicate a sce-nario where MRSA seems to be endemic and individual risk factors, namely recent hospitalizations and repeated antibiotic treatments, playa major role in the selection of drugresistant organisms. Infection control measures should be coordinated among different health care settings, and the appropriate use of antibiotics has emerged as an important issue for improving the quality of care.

Get Social With AMT

AMT 10700 W. Higgins Road, Suite 150 Rosemont, Illinois 60018

A National Certification Agency and Registry for

Medical TechnologistsMedical Laboratory TechniciansMedical Laboratory AssistantsMedical AssistantsDental AssistantsPhlebotomy TechniciansLaboratory ConsultantsAllied Health InstructorsMedical Administrative Specialists

Non-profit Org.U.S. Postage

PAIDPermit No. 1884

journal of continuing educationt iopics ssues · 2013. 4. 25. · figure 3. wide branching,...

Documents