Recently, Lablogatory interviewed R. Marshall Austin, MD, PhD, in regards to the benefits of using both liquid-based cytology and HPV testing to screen for cervical cancer. The interview below has been lightly edited for brevity and clarity.
Hi, Dr. Austin. Thanks for joining us today. Can you tells us a bit about your background?
I consider myself a gynecological pathologist, which includes surgical pathology and cytology. I’ve been involved with cervical screening issues for quite some time. Going back to the 90s, and even before that with CLIA ’88. My PhD is in virology, which is relevant now with the all the HPV issues. I did my subspecialty training in GYN and breast pathology and cytology at the armed forces institute of pathology.
Over your career you’ve authored or co-authored over 80 papers relating to cervical cancer screening. What made you so interested in this field of study?
It was an area that became a hot topic with CLIA ’88. CLIA ’88 was precipitated by a Wall Street Journal expose on Pap smear screening in the United States, which was ironic because the Pap smear has been the most effective cancer screening test in the history of medicine. This drew me in, since it was my subspecialty area of interest. There had been technological advances in the field even though the Pap smear itself hadn’t changed that much since it was introduced during World War II. Computer-assisted screening, liquid-based cytology, HPV testing all really have dramatically changed the field.
What was your initial reaction when the US preventative services task force released their draft document on cervical cancer screening recommendation in September of 2017?
I thought it was a mistake. I wrote a letter why I thought so, and apparently a lot of other people did, too.
How integral was the pathology and medical community’s reaction to this draft document in changing the USPSTF’s recommendations to include co-testing?
I’m sure that the feedback had a cumulative impact. I’ve heard different views on what components were most instrumental.
Wow! That seems really fast, considering how large your data set is.
We have kind of an unusual set up here because I work with two information scientists here at the University of Pittsburg. We automatically have all of our data being taken from our LIS into a cervical screening model which we call the Pittsburgh Cervical Cancer Screening Model and we have over 13 years of data. So we’re in kind of a unique position to very quickly put our data into different types of formats. Agnieszka Onisko, the information scientist on the publication, was able to quickly look at our data and get it into the same format as the paper from Kaiser. Once I saw how different our results actually were, my goal was to get the paper out before the USPSTF report came out. We had our tables and figures by March and I submitted the manuscript to AJCP in early May.
Let’s talk a little bit about the benefits of cotesting, and some of the downsides.
Well, the reason I always tell people, the reason that women get screened is because they don’t want to get cancer and they don’t want to die of cancer. Getting screened isn’t a pleasant experience, necessarily, but women don’t get screened because they’re worried about dysplasia or some other condition. They’re worried about cancer. The other thing that’s always been misunderstood is the limitations of screening. Screening was effectively sold to the American public by the American Cancer Society and the National Cancer Institute, and while it was an effective campaign, it basically left women with the impression that if they get screened, they won’t get cancer. Although cervical cancer screening has been the most effective cancer screening program ever, it’s never been perfect. A paper out of England in 2016 had a sophisticated analysis about the effects of cytology screening on cancer rates in England. It estimated that about 70% of cancer mortality was being eliminated with screening, and could potentially be as high as 83%, which still isn’t 100%. So when women get cancer, they get upset. My general philosophy has always been that we should do as much as we can to minimize cancer in the screened population because that’s what the public wants and expects.
The disadvantages of cotesting is one, it adds costs. Two tests cost more than one, after all. And also, cotesting adds the potential for more red flags that require potential investigation that can increase the number of procedures. Having said that, and having been involved in a number of years especially in cases where litigation is involved the public wants and expects the most protection possible. So, to me, the extra cost is still in line with what the public wants: the maximal possible protection.
A 50 year old female was admitted for acute renal failure on CKD stage IV, present with gross hematuria, anemia (due to blood loss) and hypertension. The patient has a significant history of unresolved cryoglobulinemic vasculitis initially diagnosed in 2016 and has been treated by several rounds of rituximab. Other medical histories include Sjogren’s syndrome, MGUS with monoclonal IgM Kappa, coagulopathy (protein S deficiency, on anticoagulant), hyperviscosity, myalgia, deep vein thrombosis, leg edema with superficial ulcer, pulmonary embolus and membranoproliferative glomerulonephritis (MPGN). Kidney biopsy revealed intraglomerular hyaline thrombi consistent with cryoglobulinemic glomerulopathy, interstitial fibrosis tubular atrophy, arterial sclerosis, suggestive thrombotic microangiopathy. Immunohistochemistry was positive for C3, IgM, Kappa, Lambda and CD68. Bone marrow biopsy shown dyserythropoesis without malignancy. Blood testing shown negative hepatitis panel and undetectable C4.
We observed unusual cryoprecipitate test results from this patient: gelatinous appearance precipitate which accounts for more than 40% of volume was observed in both plasma and serum and cannot be cleared at 37C° after several hours of incubation. Further testing shown incubation at 56°C for 30min cleared up the serum but not the plasma. After checking the test history, we found that there was a similar situation for the patient’s cryoprecipitate test a few months back earlier in 2018, and was reported negative for cryoglobulins due to the heat-insoluble nature of the precipitate. Patient was transfused for anemia. No plasmapheresis was done. Due to the patient’s incomplete response to rituximab, Cytoxan was also added to help improve the symptoms.
Definition: Cryoprecipitates (or cryoproteins) are blood proteins that form precipitates or gels at temperatures lower than 37°C and typically re-dissolve after warming up to 37°C. There are two types:
Cryoglobulin (CG): precipitate from both serum and plasma; either immunoglobulins or a mixture of immunoglobulins and complement components
Cryofibrinogen (CF): precipitate from plasma only; typically composed of a mixture of fibrinogen, fibrin, fibronectin, and fibrin split products
Lab Testing done in our hospital:
Blood are collected in two pre-warmed tubes (one serum, one EDTA plasma) and kept in warm water (37°C) until the serum tube clots.
The plasma and serum are extracted at room temperature, and then stored in refrigerator for 72 hours.
If cryoprotein is present, a precipitate or gel will be seen. An aliquot of the serum is rewarmed at 37°C to verify the cryo-nature.
The precipitate as a percentage of the original serum volume is measured in an ESR tube to determine the cryocrit.
Immunofixation is ordered per pathologist to identify the immunoglobulin compositions of the cryoglobulin.
Strictly speaking, cryoglobulinemia refers to the presence of cryoglobulin (CG) in a patient’s serum, which could be either asymptomatic or present with apparent clinic syndromes (i.e. cryoglobulinemic vasculitis). Cryoglobulinemia can be classified into three types (see table below ), with mixed cryoglobulinemia (type II and type III) representing 80% of the cases.
Type I cryoglobulinemia is frequently asymptomatic, while mixed cryoglobulinemia manifests clinically by a classical triad of purpura, weakness and arthralgias, as well as some other conditions including MPGN, chronic hepatitis, peripheral neuropathy, lymphoma, Raynaud’s, Sjogren’s syndrome, etc.
The presence of heat-insoluble cryoglobulins is rare, and its pathogenesis is poorly understood. On the other side, it may indicate sever clinical consequence as seen in our case and some others as mentioned above.
Essential type II cryoglobulinemia with cryoglobulin-occlusive MPGN and MGUS (Clin Chim Acta. 2009 Aug;406(1-2):170-3):79 y.o. female admitted due to edema and renal failure, cryoglobulin re-dissolved at 56°C, composed of monoclonal IgG-Kappa and polyclonal IgM.
HCV associated thrombotic microangiopathy and cryoglobulin-occlusive MPGN (Am J Med Sci. 2013 Oct;346(4):345-8):57 y.o. female, cryoglobulin re-dissolved at 47°C, composed of monoclonal IgM-Kappa and polyclonal IgG. Symptoms only partially resolved upon treatment of plasmapheresis, corticosteroids and antiviral therapy of peginterferon plus ribavirin.
Essential type I cryoglobulinemia with massive cryoglobulin-occlusive glomerulonephritis (Am J Kidney Dis. 1995 Oct;26(4):654-7):54 y.o. male progressed to ESRD prior to the detection of cryoglobulin. Cryoglobulin with white gelatinous appearance re-dissolved at 54°C, composed of monoclonal IgG-Kappa.
Primary Sjogren’s syndrome with type II cryoglobulinemia and mesangiocapillary glomerulonephritis (Nephrol Dial Transplant. 2000 Jun;15(6):917-8):82 y.o. patient with IgM-MGUS, negative BM, deposition of IgG, IgM and C3 on kidney biopsy, decreased complement levels, negative HCVAb, HBsAb, HBsAg. cryoglobulin re-dissolved at 47°C, composed of monoclonal IgM-Kappa and polyclonal IgG-Kappa.
-Rongrong Huang, PhD is a first year clinical chemistry fellow at Houston Methodist Hospital. Her interests include general clinical chemistry, genetic biochemistry and applications of mass spectrometry in clinical laboratories.
-Xin Yi, PhD, DABCC, FACB, is a board-certified clinical chemist, currently serving as the Co-director of Clinical Chemistry at Houston Methodist Hospital in Houston, TX and an Assistant Professor of Clinical Pathology and Laboratory Medicine at Weill Cornell Medical College.
This month I think it’s important to take a step back from clinical pearls, developing our interpersonal skills, and interdisciplinary dynamics and go back to what I started writing about here on Lablogatory: public health and shaping policy. (Sorry, no Transformers, Simpsons, or Star Trek this time.)
Now, you may or may not have heard in recent news that the United States Preventive Services Task Force (USPSTF) updated their long-standing guidelines for screening women for cervical cancer. Normally I wouldn’t file this away under “exciting must-read,” but I was piqued when I also read that ASCP along with the College of American Pathology (CAP), American Society of Cytopathology (ASC), American Society for Colposcopy and Cervical Pathology (ASCCP), the Society of Gynecologic Oncology, the American College of Physicians (ACP), the American Society for Cytotechnology (ASCT), the American Cancer Society (ACS) the Papanicolaou Society of Cytopathology, as well as the American College of Gynecologists (ACOG) and other professional institutions and individuals voiced concerns over the changes to the USPSTF standard.
This is a topic that can be discussed for days, but I’ll do my best to give you the readers’ digest and present the main contentions regarding this standard of patient care and laboratory methodology.
Woah. What’s going on?
Basically, because of some new research and recommendations, the USPSTF—a body which publishes the standard of care for nearly every conceivable aspect of preventive care in the US—rolled back on the algorithms for screening women for HPV and cervical cancer. It all comes down to the utilization of co-testing (doing both Pap smear cytology and HPV testing for certain age demographics) as a point of contention. Under a banner of addressing keywords like “cost” and “harm,” these new recommendations have left clinicians both in and out of the lab in stirrups—sorry, couldn’t resist that one. But don’t worry, I wouldn’t be able to track these changes or even understand them without some sort of visualization. When it comes to recommendations, standards, and guidelines I’m about as proficient as a broken manual diff counter…
Slow down. Explain co-testing and primary testing? What exactly do the old and new recommendations mean?
Okay. When women undergo routine cervical cancer screening they receive Pap smears (cytologic examinations) every three years. This testing has been the standard for a number of years and is adequately sensitive for women up to the age of 30. Often times, these younger women may have slight intraepithelial changes (LGIL) which are considered low grade and remiss on their own. After that age it has become standard practice to add the additional test (while collecting the Pap specimen) of HPV DNA testing. This adds an increased level of sensitivity/specificity and is called co-testing. The new recommendations depart from this co-testing model, citing that there are increased harms (in the form of false positives) which ultimately lead to waste and unnecessary testing for women after the age of 30. Primary testing would mean only screening with HPV DNA assays after 30. According to the National Cancer Institute, all available literature on the subject of HPV and cervical cancer testing adequately demonstrates that co-testing is the best option. A number of studies were compiled to address the harm vs. benefit of Pap and HPV testing. Together, however, these tests decrease the incidence of cervical cancer. New guidelines were made based off mathematical projections and cost-benefit analyses which try and minimize losses for screening. Dr. J. Kim, a public health researcher at Harvard, was integral in contributing models which projected the cost/benefit of changing HPV guidelines. Essentially, the study projected that, when considering “harm” (i.e. colposcopy/false negative) abandoning co-testing changed the mortality rate from 0.3-0.76 per 1000 women with co-testing, to 0.23-0.29 per 1000 women with primary HPV testing. An impressive and significant statistical advantage. However, the total number of unscreened women with mortality rates was between 1-2%. This study was a microsimulation done from historical data within rates of cytologic detection and retrospective testing data on women, projected for a future hypothetical 5 year interval. Fascinated by this study, I tried to reach out to Dr. Kim to discuss limitations in using models and simulations and public health evidence to change practices, but I’m sure she is busy and could not respond in time.
So, to co-test or not to co-test, that’s the question. Right?
In its simplest sense, yes. The major medical professional societies also publish their most current recommendations for practice standards—and the issue is that the USPSTF took a departure from what most of the professional societies recommend regarding co-testing. Late last year, the CAP, ASC, ASCT, ASCP, and the PSC issued a statement under their independent collaboration called the Cytopathology Education and Technology Consortium (or CETC). In this response to the USPSTF guideline changes, they discussed their concerns. Specifically, their objections center around the fact that without co-testing for screening, cancer prevention might be impacted negatively. The CETC claimed that sensitivity is already maximized with previously recommended co-testing guidelines. They also cite that there is only one FDA approved HPV primary screening test available—and not all labs have it! More so, CETC discussed the need to keep morphological testing continuous for women who have histories of Pap smears, the potential to overwhelm colposcopy services for screening all positive HPV patients, and the honest reality that not all clinicians would be compliant with the way the USPSTF recommends testing. The bottom line from this consortium:
Cytology and high-risk HPV co-testing should be kept as the standard screening for women aged 30-65
Primary HPV screening should only be done with validated, FDA approved testing methodologies
HPV screening methods should continue their current schedule until longitudinal data can offer new evidence for changes
So, what’s the current technological climate for how we test for HPV?
Currently, most clinicians do co-testing. The standard for Pap smears utilizes a physical tool to collect epithelial cells from the cervix at vaginal, ectocervical, and endocervical sites. The swabs are prepped on 1-2 slides, fixed with alcohol or other spray cell-preservatives and sent to labs for cytologic examination. The basic Papanicolaou staining procedure uses hematoxylin for nuclear staining, and two cytoplasmic counterstains. This is essentially a modified H&E stain to clearly visualize morphology. Staining is rarely done manually and some instruments offer stain/prep combination capability. I couldn’t find too much information on this, but I remember there not being too many official FDA approved prep machines for Pap specimens. Cytotechs and pathologists read the slides and issue sign outs on morphology according to the Bethesda system—very heavy read, don’t bother; essentially it has three main categories of normal, benign changes, and abnormal. According to ASC “for squamous lesions, TBS terminology includes atypical squamous cells of undetermined significance (ASCUS), low grade intraepithelial lesion (LGSIL or LSIL), high grade intraepithelial lesion (HGSIL or HSIL) and squamous cell carcinoma. Some laboratories also incorporate other terminologies of dysplasia and/or cervical intraepithelial neoplasia (CIN) into their reports. For glandular lesions, TBS terminology includes atypical glandular cells of undetermined significance (AGUS) and adenocarcinoma.”
As of now FDA approval for HPV primary testing for high-risk strains is limited to the Roche Cobas hrHPV test. I could link you to their website, but you’ll be sold right away. They tout the future of HPV screening is HPV primary testing and to do away with the Pap! Their graphs and figures are impressive (just like their price tag!) and there’s no doubt that sensitivity is something that real-time PCR provides more than cytologic examination. But, as always, more assays will be approved, and advancements will tweak the sensitivity and specificity higher and higher.
Got it. So, technology and lab tests are always advancing, why can’t we make this change?
It’s not so easy to change the method or assay we use to screen or diagnose patients in the lab. If you recall, I talked about how the hospital I’m currently rotating in is leading the region in advancing the new high-sensitivity troponin assays. It’s still a hard sell to many even though the data and projections seem to all point to a green light. But that’s a paradigm shift that involves side-stepping from one immunochemical assay to a more sensitive immunochemical assay. It’s the same stuff, just sharper and with more clinical data to interpret with regards to acute coronary symptoms and clinical risk stratification. Swapping an old car for a new car. This conversation is a bit more complex. The recommendations for cervical cancer screening suggest that we should move away from mostly morphologically-driven, human-based cytology interpretation and move toward PCR-based assays for detection. Literally apples to oranges. We might think we know which one is better right now, but longitudinal studies are the only way to really tease out if this change in practice to improve patient outcomes in the long run.
Where do we go from here?
Ultimately, I think a few things need to happen for this recommendation to become standard practice. First, professional societies in every discipline from gynecology to cytology need to come to an agreement. It remains to be seen whether certain agencies will adopt and recommend the USPSTF guidelines, and statements from groups like CETC reveal a vote of no confidence in this current climate. Ultimately, because of numerous objections (including the ones from ASCP and the CETC) the USPSTF does say that co-testing is still optional between patient and provider, so we’re not really in crisis mode. But what happens when the advancements and the recommendations catch up to our ability to abandon the cytologic contributions of a future useless Dr. Papanicolou? We could probably deal with that when it comes to fruition, but until then we have a real discussion about what “harm” really is. Is colposcopy flat out harm? Or are the false positives that reflex to further testing? Is the current practice a safety-net for populations across socio-economic tiers with varying access to screening in the United States? When compared to other countries, HPV prevention, vaccination, and screening is much more easily facilitated. Is this a contributing factor for our messy guidelines? Will there be more options for FDA approved methodology in the near future? There remain a number of good questions which require examining cross-sections of data and patient outcomes. And, I believe, we may see change soon—but not quite yet.
What are your thoughts? What have you experienced in your lab or clinic? Leave your comments below!
Sawaya, G. 2018. Cervical Cancer Screening—Moving from the Value of Evidence to the Evidence of Value. Journal of the American Medical Association (JAMA), Internal Medicine. doi:10.1001/jamainternmed.2018.4282
USPSTF. 2018. Screening for Cervical Cancer, US Preventive Task Force Recommendation Statement. US Preventive Task Force. Journal of the American Medical Association (JAMA) 2018;320(7):674-686. Doi:10.1001/jama/2018.10897
–Constantine E. Kanakis MSc, MLS (ASCP)CM graduated from Loyola University Chicago with a BS in Molecular Biology and Bioethics and then Rush University with an MS in Medical Laboratory Science. He is currently a medical student actively involved in public health and laboratory medicine, conducting clinicals at Bronx-Care Hospital Center in New York City.
The patient is a 48 year old male who presented after a motorcycle crash where he slide sideways into a ditch. He did not lose consciousness. At presentation he complained of pain across the abdomen, right wrist and left shoulder. He also had shortness of breath and chest wall pain. He denied hitting his head and was helmeted. He underwent exploratory laparotomy with repair of traumatic diaphragmatic hernia and left chest tube placement with post-operative course complicated by significant leukocytosis and bandemia on post-operative day 5 which triggered CT re-imaging of the chest/abdomen/pelvis. This study demonstrated intraperitoneal free air that prompted repeat exploratory laparotomy with subtotal colectomy with the abdomen open and wound vac in place. On post-operative day 9 the patient had an increasing white count that prompted return to OR with bowel staple line demonstrating leak, which prompted a small bowel resection, and ileostomy formation. After leaving the OR, the patient experienced worsening septic shock. Aggressive antibiotic therapy and IV fluid resuscitation was continued. Blood cultures taken on post-operative day 9 grew a large Gram positive rod. Growth of this organism was seen both aerobic and anaerobically.
Blood cultures showed box shaped gram positive/ variable rods that were growing on blood agar plates both aerobe and anaerobically. It was motile, indole and catalase negative, and esculin positive, and was identified by the MALDI to be Clostridium tertium.
Clostridium tertium is an aerotolerant gram positive rod that is widely distributed in the soil and can also be found the GI tract of animals and humans (1). C. tertium is non-toxin producing and produces terminal spores in anaerobic conditions (2). Infection with C. tertium is uncommon though it has been increasing in frequency as a cause of bacteremia, especially in patients with neutropenia, the immunocompromised, those with hematologic malignancy, those with inflammatory bowel disease, and in people with abdominal trauma or who have undergone abdominal surgery. Less commonly, C. tertium can be isolated in spontaneous bacterial peritonitis, enterocolitis, meningitis, septic arthritis, necrotizing fasciitis, post-traumatic brain abscess, and complicated pneumonia in mono- or polymicrobial infections (1). Additionally there has been a link between C. tertium infection and attempted suicide with the herbicides containing Glyphosate, as it causes GI toxicity which alters the gut environment (2). As with Clostridium difficile, the use of broad-spectrum antibiotics such as third-generation cephalosporins might predispose to intestinal colonization with C. tertium (1).
Increases in the diagnosis of C. tertium may be related to better diagnostic equipment such as the MALDI as it can easily be confused with a facultative anaerobic Bacillus or Lactobacillus species due to its gram variable appearance and ability to grow in aerobic conditions. A distinguishing feature between Bacillus species and C. tertium is negatively of the catalase reaction as Bacillus should be positive. Lactobacillus can still be a challenge as they are also catalase negative.
The treatment of C. tertium infection is complicated due to resistance to various antibiotics, including various beta-lactam antibiotics (such as third- and fourth-generation cephalosporins), clindamycin, daptomycin, and cotrimoxazole. Older reports state resistance to metronidazole, but this has not been confirmed in more recent publications. Available data indicate sensitivity to vancomycin, carbapenems, and quinolones (1)(3).
Salvador F, Porte L, Durán, L, Marcotti A, Pérez J, Thompson L, Noriega LM, Lois V, Weitzel T. Breakthrough bacteremia due to Clostridium tertium in a patient with neutropenic fever, and identification by MALDI-TOF mass spectrometry. International Journal of Infection Disease. 2013;17:11 (1062-1063). https://doi.org/10.1016/j.ijid.2013.03.005
You M-J, Shin G-W, Lee C-S. Clostridium tertium Bacteremia in a Patient with Glyphosate Ingestion. The American Journal of Case Reports. 2015;16:4-7. doi:10.12659/AJCR.891287.
Miller D, Brazer S, Murdoch D, Reller LB, Corey, GR. Significance of Clostridium tertium Bacteremia in Neutropenic and Nonneutropenic Patients: Review of 32 Cases. Clinical Infectious Diseases. 2001; 32:975–8
-Casey Rankins, DO, is a 1st year Anatomic and Clinical Pathology resident at the University of Vermont Medical Center.
-Christi Wojewoda, MD, is the Director of Clinical Microbiology at the University of Vermont Medical Center and an Associate Professor at the University of Vermont.
The newest generation, Generation Z, is born in the 21st century. The oldest are now 18, which means that some have started entering the work force in entry-level positions. This generation is even more comfortable with technology than Millennials, as they grew up with computers, laptops, cellphones, internet and social media all around them.
The older Gen Zers are aware of the financial crisis that occurred, which created a strong focus on saving money. This generation was brought up with a sense of “Stranger Danger” so they are concerned with their own and public safety. They have a strong family orientation and consider themselves global citizens. They are characterized by an entrepreneurial spirit, the idea that anyone can be famous, are open-minded, and care deeply about the environment.
Because of the rising cost of education, many are worried about the price of college and about saving money for their parents. It is a little too early to tell because this generation is still young, but they could have feelings of unsettlement and insecurity due to the state of the economy, environment, and world. They are very loyal, compassionate and independent and have friends around the world, even if they have never traveled abroad themselves.
-Lotte Mulder earned her Master’s of Education from the Harvard Graduate School of Education in 2013, where she focused on Leadership and Group Development. She’s currently working toward a PhD in Organizational Leadership. At ASCP, Lotte designs and facilitates the ASCP Leadership Institute, an online leadership certificate program. She has also built ASCP’s first patient ambassador program, called Patient Champions, which leverages patient stories as they relate to the value of the lab.
I think we’re embarking on an incredible generation. I interviewed someone from each of our generations about how they observed, interacted with, learned from the Generation Zs. Here are their thoughts.
The Traditionalist: Ned the Grandpa
As the grandpa of two Gen Z grandsons, Ned found them to have an expanded knowledge base of the entire world. They are sophisticated in their analysis and critical thinking because of their exposure to information that their phones and computers provide them.
Lastly, they value human diversity far more than his own generation.
The Baby-boomer: Donna the Grandma
Donna is a “Grandma Boomer” and finds the Gen Z grandchildren’s vocabulary amazing. She says they are obsessed with the mechanical stuff and are used to doing 2-3 things at the same time. They still love sports, however, it’s like a class that they study. They attend practices but still play with their friends on their computers or phones. However, they “only” text. They don’t talk on the phone.
The Gen Z’s are far more sophisticated than the Boomers, yet they can’t write or spell as well as other generations. They don’t know cursive, and the first question they ask when going somewhere is, “do they have WIFI?” Oh, and “do you have a charger?”
Another Boomer: Susan the Grandma
Susan’s greatest concern was that many high-schoolers were being treated for levels of anxiety. Why? There’s no “turn off switch” with the world. They are almost required to stay tuned to respond or react to friends 24/7. Life is all about them from Instagram to Twitter, and Snapchat and tracking the number of followers.
The GenXer, Kim the Aunt
Her nephews are definitely focused on technology. They do not like talking on the phone and prefer to only text. They have incredible access to information, but they still like to play family games because they value tradition. Her nephews are great travelers and most comfortable with airports, planes and trains, Vs. just cars or bicycles. This is attributed to their expanded world. So what’s their greatest fear? A dead battery!
Maddie the Millennial
Maddie was shocked when she noticed that her sister, who is a Gen Z, was communicating via texting with her friend who was in the same room!
-Catherine Stakenas, MA, is the Senior Director of Organizational Leadership and Development and Performance Management at ASCP. She is certified in the use and interpretation of 28 self-assessment instruments and has designed and taught masters and doctoral level students.
43 year old Vietnamese speaking man with a history of treated latent TB who presented with one month of fevers, night sweats, weight loss, and acute left facial swelling with associated pain, nasal congestion and 2 nose bleeds. The patient was found to have a polypoid mass within the left interior nasal cavity.
Biopsy Left Nasal Mass
The biopsy shows nasal mucosa with a dense submucosal lymphoid infiltrate and large areas of necrosis. The lymphocytes are somewhat pleomorphic, medium to large in size with irregular nuclear contours, vesicular chromatin and inconspicuous nucleoli. There are scattered mitoses and apoptotic cells.
By immunohistochemistry, CD3 highlights the lymphoma cells, which comprise the majority of the lymphoid infiltrate. The lymphoma cells co-express CD56 and CD7 (dim) and are negative for CD2 and CD5. The lymphoma cells also express cytotoxic markers perforin and granzyme (major subset). CD20 highlights only rare small clusters of B-cells. The lymphoma cells are also positive for EBER (Epstein-Barr Virus encoded RNA) in situ hybridization. The Ki67 (MIB1) proliferation index is 60% with focal areas exhibiting up to 80%.
Taken together, the morphologic and immunophenotypic findings are consistent with an extranodal NK/T cell lymphoma, nasal type.
Extranodal NK/T cell lymphoma, nasal type is an aggressive lymphoma that is more prevalent in Asian and South American populations. It occurs most often in adults and is more common in men than women. It is generally located in the upper aerodigstive tract, with the nasal cavity being the prototypical site. Patients tend to present in a manner similar to the patient described in this case, with rhinorrhea, pain, nasal obstruction and epistaxis due to a mass lesion. 1 The term “lethal midline granuloma” was once used to describe this entity because patients can present with locally destructive mid-facial necrotizing lesions. The early non-specific symptoms can pose a diagnostic challenge, and often result in treatment delays, which makes the aggressive disease more lethal. 2
The entity is described as NK/T cell lymphoma because although most cases are of NK-cell origin, some cases are comprised of cytotoxic T-cells. Natural killer (NK) cells are non-T and non-B lymphocytes that are part of the innate immune system. They respond immediately to antigenic challenge and are able to directly kill virally infected cells without the help of antigen presenting cells. They also secrete cytokines to increase the innate immune response. NK cells are classically positive for cytoplasmic CD3 and CD56, as well as cytotoxic molecules granzyme and perforin. Of note, NK-cells lack recombination activating gene enzymes and therefore have no clonal molecular marker for gene rearrangement such as the T-cell receptor or Immunoglobulin heavy chain. 3
Microscopically, the involved sites generally have widespread mucosal destruction. There is an angioentric and angiodestructive growth pattern that results in extensive necrosis. Another important diagnostic consideration is the very strong association with EBV. EBV is present in a clonal episomal form. This means that the infection occurs prior to and likely plays a pathogenic role in the development of NK/T cell lymphomas. 3
Following diagnosis, staging and management of the disease involves quantification of circulating EBV DNA. This can be used as a laboratory marker for disease status and progression or remission of disease. PET/CT is performed for accurate staging and patients are most commonly treated with a combination of radiotherapy and the SMILE regimen, which includes dexamethasone, methotrexate, ifosfamide, L-asparaginase and etoposide. NK/T cell lymphomas are aggressive and patients tend to have a short survival and poor overall response to therapy. 3
A recent study by Kwong, et al. showed the potential use of PD1 (programmed death ligand 1) blockade drug pembrolizumab in the treatment of relapsed or refractory NK/T-cell lymphoma. As mentioned above, the lymphocytes in this entity are invariably infected with EBV. PD1 is known to be up regulated in cells infected with EBV. In the study, seven patients who had failed treatment with the SMILE regimen were treated with pembrolizumab. After a medium follow-up of 5 months, 5 patients remained in complete remission and all patients had objective responses to treatment. 4 This shows promise as a potential new treatment for patients with this uncommon, but deadly disease.
Swerdlow SH, Campo E, Harris NL, et al. WHO Classification of Tumours of Haematopoetic and Lymphoid Tissues (Revised 4th edition). IARC: Lyon 2017.
Mallya V, Singh A, Pahwa M. Lethal midline granuloma. Indian Dermatology Online Journal. 2013;4(1):37-39. doi:10.4103/2229-5178.105469.
Tse, E, Kwong, Yok-Lam. The diagnosis and management of NK/T-cell lymphomas. Journal of Hematology and Oncology. 2017:10:85. Doi: 10.1186/s13045-017-0452-9.
Yok-Lam, K, Thomas, S.Y. Chan, Daryl Tan, et. al. PD1 blockade with pembrolizumab is highly effective in relapsed or refractory NK/T-cell lymphoma failing L-asparaignase. Blood. 2017:129:2437-2422. Doi: 10/1182/blood-2016-12-758641.
–Chelsea Marcus, MD is a third year resident in anatomic and clinical pathology at Beth Israel Deaconess Medical Center in Boston, MA and will be starting her fellowship in Hematopathology at BIDMC in July. She has a particular interest in High-grade B-Cell lymphomas and the genetic alterations of these lymphomas.
A 3 year old Caucasian male presented to the hospital due to a gastrointestinal illness and severe dehydration. Per parents, the patient started having diarrhea 4 days prior to admission. Initially, the diarrhea was watery, yellow, and foul-smelling but then became bloody and intermixed with clots. Diarrhea episodes were occurring every 20 minutes and associated with severe abdominal cramping. Additional symptoms included vomiting and malaise, but no fever was noted. There was no history of travel and no one else in the family or at daycare had similar symptoms. On physical examination, there was diffuse abdominal tenderness, signs of dehydration and altered mental status. Vitals showed an elevated blood pressure (144/97) and tachycardia (140 beats/minute). Initial lab work revealed a marked leukocytosis with left shift (78.9 TH/cm2, 62% neutrophils, 11% bands), normal hemoglobin & hematocrit, and a slight thrombocytopenia (156,000 TH/cm2). Notably, the renal panel showed kidney injury (BUN 79 mg/dL and creatinine 3.29 mg/dL) and a significantly elevated LDH (9,408 U/L). Blood and stool specimens were collected for culture and the patient was started on fluids for rehydration therapy.
The isolate was identified as Escherichia coli O157 by Vitek and latex agglutination was positive. Additionally, the patient had a GI PCR panel performed which was also positive for E. coli O157 and Shiga-like toxin-producing E. coli stx1/stx2. The specimen was sent to the department of health which also confirmed the identification of E. coli O157:H7 by molecular methods. All blood cultures remained negative. Based on the clinical presentation and culture results, the child was diagnosed with hemolytic uremic syndrome (HUS).
Escherichia coli is a gram negative rod that a member of the Enterobacteriaceae family. General features of the genus include fermentation of glucose, a negative oxidase reaction, and most strains having the ability to ferment lactose. E. coli is a normal component of gastrointestinal flora, but pathogenic strains can cause illness due to ingestion of contaminated food or water, contact with animals, or person to person spread. In the case of E. coli O157:H7, the infectious dose can be as small as 10-100 CFU.
The clinical syndromes caused by E. coli O157:H7 are commonly referred to as Shiga toxin-producing E. coli (STEC) or enterohemorrhagic E. coli (EHEC). Most commonly, E. coli O157:H7 is due to ingestion of undercooked ground beef. Presentations can range from mild non-bloody diarrhea to severe hemorrhagic colitis with hemolytic uremic syndrome (HUS). HUS is characterized by a hemolytic anemia, decreased platelets and organ damage, including renal failure and altered mental status. While only 4% of those infected with E. coli O157:H7 develop HUS, at least 80% of HUS cases in North America are due to E. coli O157:H7.
When a stool culture in received in the laboratory, a selective and differential agar to identify E. coli O157:H7 is always used. This is due to the fact that the majority E. coli O157:H7 do not ferment sorbitol. If a sorbitol negative E. coli is identified, latex agglutination is used to confirm the isolate is O157. In addition, lateral flow assays for the detection of the Shiga toxin 1 & 2 proteins are also performed as part of the routine stool culture to detect E. coli isolates that ferment sorbitol or are serotypes other than O157. Molecular testing to detect Shiga toxins provides a sensitive method of detection with an improved turnaround time. Presumptive isolates should be sent to the state department of health for confirmation and monitoring of potential outbreak situations.
Treatment for E. coli O157:H7 is generally supportive as it has been reported that certain antibiotics may stimulate further toxin production, thereby increasing the risk of HUS and as such, no routine susceptibility testing is recommended for STEC strains. Patients may need dialysis in the event of renal failure.
In the case of our patient, he had a prolonged disease course that necessitated dialysis and was complicated by a bowel perforation and pancreatitis. While his condition improved, he still requires dialysis three times a week.
-Aljunaid Alhussain, MD, is a first year Anatomic and Clinical Pathology resident at the University of Mississippi Medical Center.
-Lisa Stempak, MD, is an Assistant Professor of Pathology at the University of Mississippi Medical Center in Jackson, MS. She is certified by the American Board of Pathology in Anatomic and Clinical Pathology as well as Medical Microbiology. She is the Director of Clinical Pathology as well as the Microbiology and Serology Laboratories. Her interests include infectious disease histology, process and quality improvement, and resident education.