bluuebunny/crossref_metadata_2025_split huggingface dataset:
Myeloperoxidase Expression in B-Lymphoblastic Leukemia by Immunohistochemistry as a Diagnostic Confounder
Authors: Du Jing , Moklebust Amanda , Cherian Sindhu , Edlefsen Kerstin , Fromm Jonathan R , Li K David , Thomason Ron , Soma Lorinda 
Published: October 2020

Abstract

Background
B-lymphoblastic leukemia (BLL) is a B lineage neoplasm that expresses typical immature B cell markers, but may also express myeloid-associated antigens. Myeloperoxidase (MPO) is a heme-containing peroxidase, which has been used as the single most specific myeloid marker when assigning lineage to acute leukemia. MPO positivity by immunohistochemistry (IHC) in BLL has been historically described in a subset of patients; however, further detailed comparison of IHC to flow cytometry and IHC methodology is described herein.

Design
The University of Washington pathology database was searched for new or relapsed adult BLL from 2011–2019. Cases with blasts &gt;30% of marrow cellularity by morphology were selected. MPO IHC was performed using the Dako, polyclonal antibody (rabbit) on a Ventana instrument with streptavidin-biotin (SB) detection method. MPO IHC SB positive cases were also stained using the same antibody and platform, but a different detection method, multimer-optiview (MO). MPO IHC was called positive when &gt;10% of neoplastic blasts showed expression. MPO positive blast percentage, staining intensity and pattern were assessed. Intensity: 0=negative, 1=mild, 2=moderate, 3=bright/same level as myeloids. Cytoplasmic pattern: homogenous or granular. Concurrent flow cytometry results and cytogenetic and/or molecular studies were reviewed.

Results
35 cases were identified. Positive MPO IHC expression was present in 7/35 cases by SB method, with the percentage of MPO positive blasts ranging from 20–90% (majority &gt;70%), all ranged from 1 - 2+ intensity and most (5/7) were homogenous pattern. 4/5 MPO SB positive cases were negative by MO detection method. MPO evaluation by flow cytometry was negative in 3 of 3 cases and myeloid associated antigens were negative or low on a subset. 2/8 BLL, BCR-ABL1 cases were MPO IHC positive by SB.

Conclusion
MPO staining by IHC in BLL is present in 17% of cases, often present in the majority of blasts, and positive using the SB detection system. This aberrant staining can be negated in most cases with the MO detection system. MPO expression by flow cytometry in MPO IHC SB positive cases were negative (3/3) and were low to absent for other myeloid antigens. We agree with prior studies that MPO IHC using the SB method can be a confounder for lineage assignment in acute leukemia.
High-Volume Platelet Distribution: Who Makes the Clinical Decision?
Authors: Addams Joel , Stephens Laura 
Published: October 2020

Abstract
Anecdotally, many clinicians expect blood components to arrive in standard volumes. Blood components do not come in uniform sizes, however, and institutional experience has revealed platelet units to vary considerably in volume. Since many clinicians are not the ones examining blood products and their volumes prior to transfusion, there is a potential to transfuse a high-volume unit inadvertently to a patient at risk of transfusion-associated fluid overload (TACO), a leading cause of transfusion-related fatalities. The intent of this study was to examine the allocation practices of high-volume platelet units at an academic medical center.
Over a six-month period, blood bank technical staff prospectively logged the allocation of high-volume apheresis platelet units with volumes greater than or equal to 400 mL. The staff member who issued the product logged the reason(s), if any, why he or she selected the unit for a particular patient when more than one ABO type-specific product was available. No patient identifiers or data were logged or analyzed in this study.
Eighty-seven high-volume platelet units were recorded during the study period. The volumes ranged from 395 to 872 mL, with an average and standard deviation of 571 mL and 92 mL, respectively. Blood bank staff listed the following reasons for their allocation of high-volume platelet unit as follows: the product expiration date in 23 cases (26%), patient age in 13 cases (15%), patient sex in 12 cases (14%), and intraoperative use in 10 cases (11%).
This study enumerated the variability in platelet unit volume, with some units containing likely double or triple the volume of the ordering clinicians expectations. Blood bank technologists made clinical decisions in the allocation of high-volume units in three-quarters of the documented cases. This underscores a need to provide clearly-defined guidelines for allocating such units, as well as a mechanism for clinicians to request lower-volume units, particularly for patients at risk of TACO.
Anti-Tumor TCF1+ CD8 T Cells are Functionally Diverse and Evolve During Tumorigenesis and Progression
Authors: Schenkel Jason , Herbst Rebecca , Canner David , Li Amy , Hillman Michelle , Smith Olivia , Rozenblatt-Rosen Orit , Regev Aviv , Jacks Tyler 
Published: October 2020

Abstract
CD8+ T cells drive protective responses against infection and cancer. In the context of chronic antigen stimulation, CD8+ T cells become progressively dysfunctional, losing the ability to proliferate, secrete cytokines, and kill target cells. While dysfunctional CD8 T cells have been characterized in infection, studies examining longitudinal responses in tumors have been limited. Here, in an autochthonous model of lung adenocarcinoma using high dimensional flow cytometry and single cell RNA sequencing, we demonstrate that tumor specific CD8 T cell heterogeneity is dynamic, revealing multiple dysfunctional-like CD8 T cell populations. Among these states, we identify TCF-1+ CD8 T cells, a population previously associated with superior functionality. Temporal analysis revealed heterogeneity within the TCF-1+ CD8+ T cell compartment - SlamF6+ CD8 T cells were predominant in early stage disease, while SlamF6- CD8 T cells appeared later and were the majority during progression. Functionally, the SlamF6+ population was proliferative and expressed more inhibitory receptors than SlamF6- cells. However, SlamF6+ CD8 T cells gradually lost the ability to divide and secrete cytokines over the course of tumorigenesis, demonstrating that they become dysfunctional over time. Collectively, our results provide new insights into longitudinal response of TCF-1+ T cells over the course of tumorigenesis and have therapeutic implications for modulating the anti-tumor T cell response.
Serum Folate of Less than 7.0 ng/mL is a Marker of Malnutrition
Authors: Mattox Samantha , Kozman Diana , Singh Gurmukh 
Published: October 2020

Abstract
Folate is an essential nutrient that humans are unable to synthesize. We investigated clinical/laboratory factors associated with folate deficiency in tertiary care patients. We reviewed medical records of 1019 patients with serum folate &lt;7.0 ng/mL, 301 patients with serum folate of 15 ng/mL, and 300 patients with serum folate &gt;23 ng/mL. Serum prealbumin levels were subnormal in 54.8% of patients with serum folate &lt;7.0 ng/mL. Vitamin B12, hemoglobin, and serum albumin levels were significantly lower in the &lt;7.0 ng/mL folate group. In 62.4% of patients with serum folate &lt;7.0 ng/mL, one or more markers of malnutrition were present. The low-folate group had a significantly higher prevalence of gastrointestinal (GI) disorders, sepsis, and abnormal serum creatinine level. There were no significant differences in the two groups regarding the prevalence of diabetes; behavioral/neurological disorders, including drug and alcohol abuse; bariatric surgery; or a diagnosis of malnutrition. The average body mass index (BMI) for the &lt;7.0 ng/mL and 15 ng/mL folate groups was significantly different (28.89 and 28.31, respectively), although the difference does not appear to be clinically meaningful. The prevalence of folate deficiency depends on what is considered a normal serum folate level. Approximately 10% of tertiary care patients have levels &lt;7.0 ng/mL and exhibit other markers of malnutrition. It is recommended that patients with GI disorders, chronic kidney disease, and sepsis be routinely tested for serum folate levels, before administration of vitamin supplements. Patients with serum folate levels &lt;7.0 ng/mL should be evaluated for malnutrition, despite BMI &gt;25. Folate supplementation should be administered only after excluding coexisting vitamin B12 deficiency.
Assessment of Factor V Activity in Apheresis Platelet Units: Implications for Management of FV Deficiency
Authors: Gupta Gaurav , Hendrickson Jeanne , Bahel Parveen , Siddon Alexa , Rinder Henry , Tormey Christopher 
Published: October 2020

Abstract

Introduction
Factor V (FV) deficiency is a rare but significant disorder associated with potentially life-threatening bleeding. Management may involve plasma infusion to increase FV activity. Although the majority (~80%) of FV is present in the plasma, approximately 20% is stored in platelet (PLT) alpha granules. As such, allogeneic PLT infusion is another strategy that has been proposed in the literature to raise FV levels. Although the rationale for this approach is understandable, these recommendations are based largely on anecdotal reports. Since FV is the most labile coagulation factor from an in vitro standpoint and given that allogeneic PLTs are stored at room temperature for 5–7 days after collection, we hypothesized that there would be low FV activity in PLT units and tested our hypothesis by assessing FV levels across PLT products.

Material and Methods
PLTs were aliquoted from apheresis units only, including those with PLT additive solution (PAS) and from non-PAS units (N=26) on post-collection days 3–6. Supernatant was prepared from PLTS by centrifugation (5000 rpm X 5 min) and was stored at -80oC until analyzed. PLT lysate (PL) was prepared by freezing (-80oC) then thawing (370 C) followed by centrifugation (10,000 rpm X 10 min). FV activity was tested in the PLT unit supernatant and lysate using a clinical-grade assay based on prothrombin time correction of FV deficient plasma and expressed as percent activity; reference range for normal plasma FV levels at our facility is 50–150% activity. Statistical analysis was performed using student t test. The data were expressed as mean ± SD (P-value &lt;0.05 considered significant).

Results
FV activity was markedly low in PAS PLT units, in both supernatant (3.70% ± 1.02%) as well as PL (3.26% ± 1.02%); N=16. FV activity in non-PAS PLT units (N= 10) was also somewhat low relative to reference range plasma levels in both supernatant (44.55% ± 6.46%) and PL (39.67% ± 6.33%); however, it was significantly higher (P &lt; .001) when compared to PAS PLT units. FV activity from aliquots extracted from the same PAS PLT units and examined serially over storage time (N=3) showed FV levels on day 3 of storage (supernatant 5.03% ± 1.41%; PL: 3.89% ± 1.03%) to be not significantly different than levels measured on day 5 of storage (supernatant: 3.10% ± 0.57%; PL: 2.61% ± 0.41%; P=0.4).

Conclusion
FV is substantially depleted across PLT components, with particularly low levels observed in PAS PLTs. Given comparable supernatant and PL FV levels, there appears to be little FV reserve available in PLT lysate. Based on these data, plasma should be considered a first-line therapy for most non-complex FV deficiency-associated bleeds. If PLTs are considered for additional use, then non-PAS PLT units should be preferentially employed.
Del20q and Gain/Loss of Sex Chromosomes in Patients with Plasma Cell Neoplasm
Authors: Lee Benjamin , Li Shiyong 
Published: October 2020

Abstract

Aim
Plasma cell neoplasm (PCN) is a clonal proliferation of plasma cells involving bone marrow and extramedullary anatomic sites. The purpose of this study is to investigate the prevalence and clinical significance of del20q and gain/loss of sex chromosomes in patients with PCN.

Materials and Methods
Routine karyotype analysis was performed on bone marrow aspirate specimens from patient with plasma cell neoplasm in the cytogenetics laboratory using the G-banding technology. Results from January 1, 2012 to June 21, 2019 were retrieved for analysis.

Results
Among the 8,665 karyotype results, 1,116 were abnormal and 593 demonstrated del20q and/or gain/loss of sex chromosomes. 33% (113/347) have bone marrow PCN when del20q and gain/loss of sex chromosomes occur in isolation, while 94% (256/250) have bone marrow PCN when 1 or more other autosomal abnormalities are also present. In those with isolated del20q or gain/loss of sex chromosomes, the frequency of bone marrow PCN ranges from 29–50%. Del20 occurs 1–6 years after therapy and in some patients is transient. Gain/loss of sex chromosome is generally present during the follow-up period.

Conclusion
Del20q and gain/loss of sex chromosomes do not appear to correlate with bone marrow PCN. Isolated Del20q likely represents transient treatment-related abnormality, while isolated gain/loss of sex chromosomes is probably age-related.
Lipemic Interference in Basic Metabolic Panels: Increasing the Lipemia Index Threshold in Order to Decrease the Frequency of Ultracentrifugation
Authors: Krenzer Joseph , Nelson Alyson , Robakowski Trisha , Grant Kevin , Galior Kornelia , Hackenmueller Sarah A 
Published: October 2020

Abstract

Introduction
Lipemia in clinical chemistry samples is a problematic form of interference. Clearing these samples for routine testing can be time consuming and increases the turn-around time for these specimens. In our laboratory, samples with a lipemia index &gt;50 (L-index) are manually inspected and visibly lipemic specimens are cleared by ultracentrifugation.

Objective
The objective of this study was to determine at what L-index ultracentrifugation of lipemic BMP specimens is necessary prior to sample testing to ensure accurate results.

Methods
Specimens consisted of routinely ordered basic metabolic panels (BMP) that met current criteria for ultracentrifugation, which included an L-index &gt;50 as measured on the Abbott Architect c8000 and visual lipemia. Specimens meeting these criteria were ultracentrifuged and retested. The difference of the pre-ultracentrifuged and post-ultracentrifuged result was evaluated and put into a percent to find the ‘percent difference’ and evaluated against the total allowable error (TEa) for each analyte. If the difference observed following ultracentrifugation was less than or equal to 50% of the TEa, clearance of lipemia by ultracentrifugation was considered unnecessary. Values from all BMP component tests were analyzed in order to find an L-index threshold at which samples need to be ultracentrifuged which could be applied to the entire panel. The report of lipemic indices for BMPs for the month of January 2020 were extracted from the laboratory information system to evaluate the potential impact of altering the L-index threshold for ultracentrifugation.

Results
Based on the acceptance criteria of ≤50% of TEa, L-index thresholds for Na, K, Cl, calcium, glucose, creatinine, CO2 and BUN were &lt;203, &lt;410, &lt;287, &lt;387, &lt;410, &lt;285, &lt; 153 and &lt;285, respectively. All the calculated differences or percent differences for each analyte did not exceed 50% of the TEa for a given analyte when the L-index was 150 or less. Adjusting the L-index to 150 and applying it to the 195 lipemic BMP samples in January 2020, would have potentially decreased the number of samples requiring ultracentrifugation to 24 lipemic BMPs (88% reduction).

Conclusion
These data suggest that an L-index greater than 150 can be used for all analytes within a BMP as the threshold for requiring ultracentrifugation. The BMP is one of the most frequently ordered tests in our laboratory and consistently accounts for a substantial portion of the lipemic samples that require ultracentrifugation. Increasing the L-index at which samples will be ultracentrifuged from 50 to 150 would potentially result in an 88% reduction in one month of BMP samples requiring ultracentrifugation.
Development and Validation of Leukocyte Enzyme Activity Assay by LC-MS/MS for Diagnosis of Krabbe Disease and Other Lysosomal Storage Diseases
Authors: Liao Hsuan-Chieh , Mitchell Laura , Sadilkova Katerina , Dickerson Jane , Jack Rhona , Anna Scott 
Published: October 2020

Abstract

Background
Deficiency of the lysosomal enzyme galactosylcerebrosidase (GALC) causes Krabbe disease. The diagnosis for Krabbe disease includes measurement of GALC enzymatic activity by radioisotope assay or accumulation of metabolite psychosine. To improve current diagnostic workflow and assay performance, we developed and validated a leukocyte enzymatic assay by using liquid chromatography tandem mass spectrometry (LC-MS/MS) for lysosomal storage diseases.

Methods
Leukocytes were separated and extracted from whole blood samples, and total protein was quantitated by BCA method. Commercialized and multiplexed substrates, internal standards, and buffer were incubated with cell lysates. The lysosomal enzymes in leukocytes metabolized the artificial substrate into product which is structurally identical to the internal standard. Liquid-liquid extraction was performed and supernatant was dried down and reconstituted. Liquid chromatography separation was achieved by Waters CSH C18, 2.1 x 50 mm column and Acquity UPLC system. A Waters Xevo TQS tandem mass spectrometer was used for mass detection.

Results
Enzymatic reaction products for six lysosomal enzymes were chromatographically resolved from substrate breakdown products through 3.5 minutes gradient liquid chromatography. Intra-assay imprecision was determined by 11 replicates of samples containing low and high concentration (CV&lt;15%). Carryover was determined by assaying triplicates of cell lysate-free cocktails directly after injection of high enzyme activity sample (less than 0.1%). No matrix effect was found. The GALC enzyme activity was calculated and standardized by corresponding product and internal standard ratios from 5-point standard curve. The range of enzyme activity from three, known affected patients is 0.01–0.07 (nmol/hr/mg protein); whereas, two identified carriers had enzyme activate in the range of 0.14–0.40 (nmol/hr/mg protein). The reference interval was established from 63 residual, unaffected samples and was 0.12–5.97 (1.44±1.44) nmol/hr/mg protein.

Conclusions
A simple and multiplexed LC-MS/MS assay was developed which can measure small amounts of residual GALC enzyme activity in leukocytes. This confirmatory assay will aid in the diagnosis and prognosis (i.e. differentiate disease severity) of Krabbe disease and other lysosomal storage disorders.
Synovial Metaplasia of the Endometrium
Authors: Yurtsever N , Truskinovsky A 
Published: October 2020

Abstract

Introduction/Objective
Synovial-like metaplasia is well known to accompany breast implants, but it can also rarely be found in the endometrium, associated with intrauterine devices. We present two cases of endometrial synovial-like metaplasia.

Methods
The hysterectomy specimens were diagnosed at Northwell Health and reviewed at the departmental Consensus Conference. Immunostains for cytokeratin AE1/AE3 were performed on the endometrium. The histopathologic findings were correlated with the clinical information.

Results
Both patients had had intrauterine devices placed for birth control. One, a 47-year-old female with a family history of Lynch syndrome and newly diagnosed with a corresponding mutation, underwent prophylactic hysterectomy. The second patient, a 50-year-old female with a history of uterine prolapse, was experiencing stress urinary incontinence, for which she underwent supracervical hysterectomy. Both patients had intrauterine devices in place at the time of hysterectomy.
Both uteri showed endometrial synovial-like metaplasia. It was associated with a complete absence of the endometrial surface epithelium. The synovial-like cells were eosinophilic, plump and elongated, were palisaded perpendicular to the endometrial surface and were negative for cytokeratin AE1/AE3, which immunostain also confirmed the absence of the endometrial surface epithelium and the presence of some residual deeper endometrial glands.

Conclusion
Endometrial synovial-like metaplasia is a rare entity described in patients with intrauterine devices. The mesodermal-derived endometrium is apparently able, when exposed to a semi-mobile foreign body, to develop a way to minimize friction by mimicking synovium, similar to the connective tissue facing breast implants. Such surface must be better at withstanding chronic friction than the regular endometrial epithelial surface, leading to this unusual and fascinating change. Since the resulting surface mimics synovium perfectly, both morphologically and immunohistochemically, and serves a very similar mechanical purpose, we propose to call this phenomenon, wherever it is found, “synovial metaplasia” instead of the current term “synovial-like metaplasia.”
Primary Intestinal-type Adenocarcinoma of the Vulva Causing Secondary Extramammary Paget Disease
Authors: Forsythe D J , Weisman P 
Published: October 2020

Abstract

Casestudy
A 65-year-old woman with a known history of anogenital extramammary Paget disease presented with a pedunculated lesion arising from the vulvar vestibule near the urethral meatus, discovered during a routine surveillance exam.

Results
Biopsy of the lesion revealed an adenocarcinoma with villoglandular architecture and pseudostratified columnar epithelium with goblet cells that is arising in direct continuity with the uninvolved squamous mucosa. In addition to the villoglandular component, an area of dyscohesive cells with signet ring morphology within a mucinous background was present. Biopsy of a lesion inferior to the index mass showed Paget disease with similar morphology to the invasive adenocarcinoma. By immunohistochemistry (IHC), the invasive adenocarcinoma was diffusely positive for CK20, CK7, and CDX-2 with patchy staining for p16. Metastasis was excluded by colonoscopy, mammography, and PET-CT. Additional IHC stains for GCDFP-15, GATA-3, PAX-8, S100, p63, and PSA were all negative on subsequent biopsies performed for mapping the extent of disease. These features supported a diagnosis of primary intestinal-type adenocarcinoma of the vulva and pointed to this tumor as a source of her Paget disease.

Conclusion
Primary intestinal-type adenocarcinomas of the vulva are extremely rare with only a small number of case reports in the literature. It is suggested that these tumors acquire a colorectal phenotype due to the cloacal origin of the vulvar vestibule. No standard treatment approach has been developed due to the rarity of this disease, but the majority of reported cases have had an indolent course. To the best of our knowledge, other than this case, there is only a single additional case report in the literature of secondary Paget disease arising from this tumor type. We underscore the propensity of this tumor to cause secondary Paget disease, mimicking spread from a colorectal primary site.