Deep Learning Provides Rapid Screen for Breast Cancer Metastasis with Sentinel Lymph Nodes.

OBJECTIVE: Deep learning has been shown to be useful in detecting breast cancer metastases by analyzing whole slide images (WSI) of sentinel lymph nodes; however, it requires extensive analysis of all the lymph node slides. Our deep learning study attempts to provide a rapid screen for metastasis by analyzing only a small set of image patches to detect changes in tumor environment. METHODS: We designed a convolutional neural network to build a diagnostic model for metastasis detection. We obtained WSIs of Hematoxylin and Eosin-stained slides from 34 cases with equal distribution in positive/negative categories. Two WSIs were selected from each case for a total of 69 WSIs. From each WSI, 40 image patches (100x100 pixels) were obtained to yield 2720 image patches, from which 2160 (79%) were used for training, 240 (9%) for validation, and 320 (12%) for testing. Interobserver variation was also examined among 3 users. RESULTS: The test results showed excellent diagnostic results: accuracy (91.15%), sensitivity (77.92%), and specificity (92.09%). No significant variation in results was observed among the 3 observers. CONCLUSION: This preliminary study provided a proof of concept for conducting a rapid screen for metastasis rather than an exhaustive search for tumors in all fields of all sentinel lymph nodes.

Technical Note: Bimodal Negative Interference of Biotin in Roche IL-6 Immunoassay.

OBJECTIVE: Interleukin -6 (IL-6) is an important diagnostic test in COVID-19 patients to determine whether to initiate tocilizumab therapy or mechanical ventilation. We investigated potential interference of biotin in Roche IL-6 assay which utilizes biotinylated antibody. METHODS: We prepared three serum pools from left-over specimens which showed IL-6 values over 40 pg/mL. Then aliquots of each serum pool were further supplemented with various amounts of biotin expected in patients taking biotin supplement and then IL-6 values were measured again using Roche IL-6 assay on the Cobas e411 analyzer. RESULTS: We observed negative interference of biotin in IL-6 assay but interference was bimodal as maximum negative interference was observed with 100 ng/mL biotin but not with 1000 ng/mL. However, no interference was observed in the presence of 25 ng/mL biotin. CONCLUSIONS: Biotin showed negative interference with IL-6 assay.

Obesity Is a Risk Factor Associated with H. pylori-negative MALT Lymphoma of Stomach.

OBJECTIVE: More than 90% of gastric mucosa-associated lymphoid tissue (MALT) lymphomas are attributed to Helicobacter pylori infections. However, the pathogenesis of H. pylori-negative MALT lymphomas is controversial, and additional etiologies need to be investigated. MATERIALS: A retrospective study of gastric MALT lymphoma cases over a 15-year period revealed 56 cases. The H. pylori status, clinical information, and body mass index (BMI) data were collected. The results of the urea breath test, serology, stool antigen, and previous biopsy results were documented. RESULTS: The 56 cases had an average height of 166.57 cm (range, 147.3-190.5), weight of 83.98 kg (range 55-153.1), and body mass index (BMI) of 30.34 kg/m2 (range, 17.96-49.77). Twenty-one cases were H. pylori-positive (37.5%), with a mean BMI of 27.36 kg/m2 (range, 17.96-47.25), and BMI>30 kg/m2 in 5 (23.8%) patients. Thirty-five cases were H. pylori-negative, with a mean BMI of 31.90 kg/m2 (range, 18.17-49.77), and 20 (57.1%) having BMI>30 kg/m2. A Fisher's exact test and two-tailed test showed a statistically significant difference between the two groups. CONCLUSION: Obesity leads to a baseline state of chronic inflammation and increased production of pro-inflammatory cytokines that can stimulate the lymphocytes, leading to lymphomatous proliferation. Our study suggests a potential correlation between obesity and the risk of development of primary gastric MALT lymphoma.

Pathology Residency Curriculum: Time for a Change?

The current Anatomical and Clinical Pathology residency curriculum, as outlined by the American Board of Pathology (ABP), emphasizes resident exposure to a wide variety of subjects without in-depth training. This has led to a large number of residents pursuing fellowship training. With the demand for further sub-specialization, there is a necessity for the establishment of an updated curriculum that not only encompasses the basic knowledge of pathology but is also focused on training residents in their desired subspecialty.We herein propose a new comprehensive AP/CP residency syllabus. The new curriculum will be divided into two major categories: preliminary and subspecialty training. The curriculum will require residents to undergo basic pathology training within the first two preliminary years, followed by two subspecialty years. In their subspecialty years, each resident will be required to either pick two subjects as majors, each having a duration of one year, or one subject as a major and two subjects as minors, in which case the major will have a one-year duration and the minors will each be six months in length. The proposed curriculum meets the current guidelines of the ABP, reduces the burden of residents to complete multiple fellowships, and allows residents earlier entrance into the workforce.

COVID-19 Cardiac Injury: An Important Cause of COVID-19 Related Morbidityand Mortality.

OBJECTIVE: Although coronavirus disease 2019 (COVID-19) typically presents as a respiratory illness, co-existent cardiovascular symptomatology associated with an elevated serum troponin level has been identified as a risk factor for adverse outcomes. Our study addressed the need to correlate serum cardiovascular biomarkers with tissue pathology based on autopsy. MATERIALS AND METHODS: In 13 patients, we reviewed the clinical history and measurements of serum troponin and other biomarkers and correlated them with autopsy findings. RESULTS: At autopsy, the 13 COVID-19 patients exhibited evidence of diffuse alveolar damage (DAD) and cardiomegaly (heart weights ranged from 380 to 1170 grams). Of the 13 patients, three had elevated troponin I and evidence of severe coronary artery disease (CAD) (cases 4, 5, and 11), while six had elevated troponin I without evidence of severe CAD (cases 1, 3, 6, 7, 8, and 9), and four had no clinical or pathological evidence of CAD. Of note, cases 7 and 9 had significantly elevated troponin I levels (8.84 ng/mL and 4.94 ng/mL, respectively). Several cases showed focal degenerative change or damage of cardiomyocytes. However, none of the cases had evidence of lymphocytic myocarditis. CONCLUSION: Although we observed elevated biomarkers of heart failure in some cases, it was not a consistent finding and did not correlate with evidence of myocarditis. The elevated biomarkers may reflect non-ischemic heart damage as a consequence of COVID-19 infection.

Acute myeloid leukemia with mutated NPM1 mimics acute promyelocytic leukemia presentation.

OBJECTIVES: To investigate clinicopathological and molecular features of NPM1-mutated acute myeloid leukemia that presented with infrequent acute promyelocytic leukemia (APL)-like phenotype and clinical presentation. METHODS: Cases with both de novo or secondary Acute Myeloid Leukemia (AML) were retrieved. Data from flow cytometry immunophenotyping, cytogenetics, molecular studies, and clinical presentation were analyzed. RESULTS: Cases presented with abnormal coagulation parameters and low platelets count; four of them showed a DIC index compatible with overt DIC. Two cases showed Auer rods. In all cases, immunophenotypes mimicked APL: blasts expressed CD33, CD13, and cytoplasmic MPO but did not express CD34, HLA-DR, or CD11b. Notably, CD4 expression was observed in all cases. Neither t(15;17) nor PML/RARα gene rearrangement was detected. NPM1 gene mutation was identified in all cases. In four cases, TET2 or IDH2 co-mutations were identified. CONCLUSIONS: Our findings provide additional evidence of association between NPM1-mutated AML with TET2 or IDH2 co-mutations and the APL-like immunophenotype. This AML subset was found to exist in both de novo and secondary AML. High WBC count and blasts with low to moderate side scatter and significant expression of CD4 are observed features that could assist in the differential diagnosis with APL. The occurrence of significant elevated D-dimer levels, or even overt DIC observed at diagnosis in these cases could be relevant for this AML subtype.

HIV-associated plasmablastic lymphoma in the era of HAART: a single-center experience of 21 patients.

OBJECTIVES: Patients with HIV infection have an increased risk of developing plasmablastic lymphoma (PBL). In this study, we reviewed the clinicopathologic features of PBL in HIV+ patients in the era of HAART from a single health center. DESIGN: Retrospective study. METHODS: The morphologic, immunophenotypic, and clinical features were reviewed in these HIV+ patients with PBL and univariate analysis was employed to determine the survival prognosis. RESULTS: During the interval of 1 January 2008 to 30 December 2018, we identified 95 HIV+ patients with aggressive non-Hodgkin B-cell lymphomas. Among these patients, there were 21 (22%) patients with PBL (19 men and two women; median age: 45 years). Seven patients had PBL at their initial HIV diagnosis and 14 developed PBL after a median interval of 7.7 months of HIV diagnosis. Lymph nodes (n = 10), oral cavity/sinonasal mass (n = 6), and rectal masses (n = 5) were the common involved sites, and five of 15 (33%) had bone marrow involvement. Lymphoma cells were immunoreactive for MUM-1/IRF4 (100%), CD138 (90%), CD45 (63%), CD79a (47%), and CD30 (25%). Proliferation rate assessed by Ki67 was at least 90% in 18 of 20 cases. Eighteen patients received chemotherapy including etoposide, prednisone, vincristine, cyclophosphamide, and doxorubicin (n = 13) and cyclophosphamide, doxorubicin, vincristine, and prednisone (n = 2). With a median follow-up time of 19 months, nine out of 17 patients died. Bone marrow involvement was associated with a poorer overall survival (median: 4.7 months, P = 0.015). CONCLUSION: PBL is the second most common type of aggressive lymphoma and often presents in lymph nodes of patients with poorly controlled HIV infection. Bone marrow involvement is associated with a poorer outcome.

Educational Case: Mantle Cell Lymphoma.

The following fictional case is intended as a learning tool within the Pathology Competencies for Medical Education (PCME), a set of national standards for teaching pathology. These are divided into three basic competencies: Disease Mechanisms and Processes, Organ System Pathology, and Diagnostic Medicine and Therapeutic Pathology. For additional information, and a full list of learning objectives for all three competencies, see http://journals.sagepub.com/doi/10.1177/2374289517715040.1.

Cryptic Acute Promyelocytic Leukemia (APL) Presenting as Seizures in an Adolescent.

There are approximately 800 new cases of acute promyelocytic leukemia (APL) in the United States every year. APL is rarely observed in pediatric populations, and accounts for less than 5-10% of all pediatric cases of acute myeloid leukemia (AML). APL typically presents with symptoms related to the pancytopenia such as fatigue due to anemia, bleeding and bruising secondary to thrombocytopenia, and infections attributed to a lack of functioning leukocytes. The presentation of APL in the central nervous system (CNS) is a rare phenomenon. In addition, APL has a dismal prognosis when found in the CNS. In this case study, we describe a unique presentation of pediatric APL with cryptic insertion of the promyelocytic leukemia (PML)-retinoic acid receptor-alpha (RARA) fusion protein with a surprisingly excellent recovery.

Distortions but not Definitive Peaks by Capillary Serum Protein Electrophoresis Indicate Monoclonal Gammopathy of Undetermined Significance rather than Multiple Myeloma.

OBJECTIVE: Capillary electrophoresis of serum proteins demonstrates occasional distortions. Distortions or peaks in the gamma, beta, and alpha-2 zones may represent monoclonal gammopathy. In this study, we investigated if such distortions are associated with monoclonal gammopathy of undetermined significance (MGUS) or multiple myeloma. METHODS: Consecutive serum protein electrophoresis results were reviewed and immunofixation studies were recommended on specimens exhibiting distortions or distinct peaks in the gamma, beta or alpha-2 zones. RESULTS AND DISCUSSION: Of the 471 cases, we observed distortions in 101 cases. In the immunofixation studies, 17.8% of cases had a diagnosis of MGUS, but none contained multiple myeloma. CONCLUSIONS: We conclude that distortions in serum capillary electrophoresis may be associated with MGUS, but not multiple myeloma.

Overutilization of Test for Hemoglobinopathy Evaluation: Experience from a Tertiary Care Academic Medical Center.

BACKGROUND: Hemoglobin electrophoresis is a common clinical laboratory test for the identification of hemoglobinopathies in clinical practice. We investigated the utilization of this test in our academic teaching hospital and hypothesized that hemoglobin electrophoresis evaluation is overutilized at this institution. METHODS: 128 consecutive cases were analyzed and their medical records were studied to determine the clinical indication for the hemoglobin electrophoresis. RESULTS: Of the 128 cases studied, only 44% of cases had a justifiable reason for obtaining the hemoglobin electrophoresis, whereas the remaining 56% of cases did not have an acceptable indication for obtaining the test. CONCLUSIONS: We conclude that hemoglobin electrophoresis is overutilized in our academic medical center and we recommend consultation with clinical pathologists prior to ordering such tests.

Automated Diagnosis of Lymphoma with Digital Pathology Images Using Deep Learning.

Recent studies have shown promising results in using Deep Learning to detect malignancy in whole slide imaging, however, they were limited to just predicting a positive or negative finding for a specific neoplasm. We attempted to use Deep Learning with a convolutional neural network (CNN) algorithm to build a lymphoma diagnostic model for four diagnostic categories: (1) benign lymph node, (2) diffuse large B-cell lymphoma, (3) Burkitt lymphoma, and (4) small lymphocytic lymphoma. Our software was written in Python language. We obtained digital whole-slide images of Hematoxylin and Eosin stained slides of 128 cases including 32 cases for each diagnostic category. Four sets of 5 representative images, 40x40 pixels in dimension, were taken for each case. A total of 2,560 images were obtained from which 1,856 were used for training, 464 for validation, and 240 for testing. For each test set of 5 images, the predicted diagnosis was combined from the prediction of five images. The test results showed excellent diagnostic accuracy at 95% for image-by-image prediction and at 100% for set-by-set prediction. This preliminary study provided a proof of concept for incorporating automated lymphoma diagnostic screen into future pathology work-flow to augment the pathologists' productivity.

Fatal Intracranial Hemorrhage in a Young Pregnant Patient with Acute Promyelocytic Leukemia.

Acute promyelocytic leukemia during pregnancy is associated with a high risk of fetal and obstetrical complications. Although the prognosis of acute promyelocytic leukemia is favorable due to disease-specific drugs, such as all-trans retinoic acid, early death due to fatal intracranial hemorrhage has been observed in some cases. In this study, we present a case of catastrophic intracranial hemorrhage in a young pregnant patient with the finding of leukemic involvement of the placenta. To our knowledge, this is the first confirmed case of acute promyelocytic leukemia involving the placenta. The clinical history, pertinent histological findings, and clinical outcomes will be discussed.

Proteomics Analysis of FLT3-ITD Mutation in Acute Myeloid Leukemia Using Deep Learning Neural Network.

Deep Learning can significantly benefit cancer proteomics and genomics. In this study, we attempted to determine a set of critical proteins that were associated with the FLT3-ITD mutation in newly-diagnosed acute myeloid leukemia patients. A Deep Learning network consisting of autoencoders formed a hierarchical model from which high-level features were extracted without labeled training data. Dimensional reduction reduced the number of critical proteins from 231 to 20. Deep Learning found an excellent correlation between FLT3-ITD mutation with the levels of these 20 critical proteins (accuracy 97%, sensitivity 90%, and specificity 100%). Our Deep Learning network could hone in on 20 proteins with the strongest association with FLT3-ITD. The results of this study allow for a novel approach to determine critical protein pathways in the FLT3-ITD mutation, and provide proof-of-concept for an accurate approach to model big data in cancer proteomics and genomics.

Howell-Jolly Body-Like Inclusions in Neutrophils of Patients With Myelodysplastic Syndrome: A Novel Correlation.

CONTEXT.­: Howell-Jolly body-like inclusions have been previously associated with patients who are human immunodeficiency virus (HIV) infected, taking antiviral medications, and immunosuppressed. These inclusions appear in neutrophils and resemble Howell-Jolly bodies of normoblasts in abnormal erythropoiesis. In granulocytes, they are thought to represent detached nuclear fragments produced from dysplastic granulopoiesis. To the best of our knowledge, no association of Howell-Jolly body-like inclusions and myelodysplastic syndrome (MDS) has been reported previously. OBJECTIVE.­: To establish an unprecedented correlation of Howell-Jolly body-like inclusions in patients with MDS. DESIGN.­: Eleven bone marrow cases from patients diagnosed with MDS and 20 bone marrow cases with no significant pathologic alterations were retrospectively reviewed. Detailed medical record review was performed to ensure none of the patients had a history of HIV, or was taking immunosuppressants and/or antiviral medications. RESULTS.­: Eight of 11 cases (72%) from the study group show detached intracytoplasmic inclusions in a minority (<5%) of mature neutrophils consistent with Howell-Jolly body-like inclusions. No Howell-Jolly body-like inclusions were identified in the control group. Notably, none of the selected patients had a history of HIV or was taking immunosuppressants and antiviral medications. CONCLUSIONS.­: In review of the literature, Howell-Jolly body-like inclusions seem to correlate with immunosuppression and antiviral therapies with nucleoside analogs. We propose that the formation of Howell-Jolly body-like inclusions is the consequence of dysplasia, and hence its correlation not only with the abovementioned conditions, but also with MDS. The inclusions are, however, seen in only a minority of white cells (<5%), which is probably why they were not brought to practicing pathologists' awareness in the past. This study aims to raise awareness and correlate the presence of Howell-Jolly body-like inclusions in MDS.

PIFA PLUSS P4 Assay for Screening of Heparin-Induced Thrombocytopenia.

BACKGROUND: The PIFA PLUSS PF4 Rapid Assay (PIFA) is a rapid screening test used for the diagnosis of heparin-induced thrombocytopenia (HIT). OBJECTIVE: To determine the usefulness of this assay as a screening method in our institution. METHODS: A total of 159 specimens from patients with suspected HIT were included in our study. We simultaneously performed PIFA assay and confirmatory polyspecific enzyme-linked immunosorbent assay (ELISA). We subjected most of the specimens with false-negative results to serotonin release assay (SRA). RESULTS: The initial sensitivity and specificity of the PIFA assay were calculated as 27.3% and 71.5%, respectively. A total of 12 of 16 false-negative results were further tested using the SRA method. The revised sensitivity and specificity were 50.0% and 73.5%, respectively. CONCLUSIONS: Despite its appealing feature of yielding rapid results, the PIFA assay is inadequate as a sole screening test for HIT because of its high probability of missing many true cases of HIT.

Taking Advantage of Assay Harmonization, Biotin Interference in the LOCI Digoxin Assay Could Be Eliminated by Using the ADVIA Centaur Digoxin Assay.

Biotin at elevated concentration interferes with immunoassays that utilize biotin in assay design. We earlier reported interference of biotin in the luminescent oxygen channeling assay (LOCI) digoxin assay which utilizes biotinylated antibody against digoxin. However, the ADVIA Centaur digoxin assay, also manufactured by Siemens Diagnostics, does not utilize biotin in assay design. We hypothesized that if the LOCI and the ADVIA Centaur digoxin assay are harmonized, then interference of biotin in the LOCI digoxin assay could be eliminated by using the ADVIA Centaur digoxin assay. We analyzed 25 specimens from patients receiving digoxin using both assays to investigate harmonization between these two assays. Then aliquots of drug-free serum pool were supplemented with various biotin concentrations (range: 10 ng/mL to 2000 ng/mL) followed by measuring apparent digoxin levels using the ADVIA Centaur digoxin assay. In another set of experiments, aliquots of a serum digoxin pool were supplemented with biotin (10-2000 ng/mL) and digoxin concentrations were measured by the ADVIA Centaur digoxin assay. We observed an excellent correlation between digoxin values obtained by the LOCI digoxin assay (reference method) and the ADVIA Centaur digoxin assay (y= 1.0514 x+0.1083, r=0.99) indicating that both assays are harmonized. We did not observe any interference of biotin even at a highly elevated concentration of 2000 ng/mL with the ADVIA Centaur digoxin assay. We conclude that taking advantage of assay harmonization, interference of biotin in the LOCI digoxin assay can be eliminated by using the ADVIA Centaur digoxin assay.

Can Tranexamic Acid Reduce Blood Loss during Major Cardiac Surgery? A Pilot Study.

We examined the effectiveness of tranexamic acid in preventing intraoperative blood loss during major cardiac surgery. Out of initial 81 patients undergoing major cardiac surgery (both coronary artery bypass and valve repair procedures) at our teaching hospital, sixty-seven patients were selected for this study. We compared estimated blood loss, decrease in percent hemoglobin and hematocrit following surgery between two groups of patients (none of them received any blood product during surgery), one group receiving no tranexamic acid (n=17) and another group receiving tranexamic acid (n=25). In the second study, we combined these patients with patients receiving modest amounts of blood products (1-2 unit) and compared these parameters between two groups of patients (25 patients received no tranexamic acid, 42 patients received tranexamic acid). In patients who received no blood product during surgery, those who received no tranexamic acid showed statistically significant (independent t-test two tailed at p<0.05) reduced estimated blood loss (mean: 713.5 mL, SD: 351.6, n=17) compared to those who received tranexamic acid (mean: 987.2 mL, SD: 459.9, n=25). We observed similar results when the patients receiving no blood products and patients receiving modest amount of blood products were combined based on the use of tranexamic acid or not. No statistically significant difference was observed in percent reduced hemoglobin or hematocrit following surgery in any group of patients. We conclude that intraoperative antifibrinolytic therapy with tranexamic acid does not reduce intraoperative blood loss during major cardiac surgery which contradicts popular belief.