Showing papers in "American Journal of Clinical Pathology in 2008"

Automated blood cell counts: state of the art.

Abstract: The CBC count and leukocyte differential count (LDC) are among the most frequently requested clinical laboratory tests. These analyses are highly automated, and the correct interpretation of results requires extensive knowledge of the analytic performance of the instruments and the clinical significance of the results they provide. In this review, we analyze the state of the art regarding traditional and new parameters with emphasis on clinical applications and analytic quality. The problems of some traditional parameters of the CBC count, such as platelet counts, some components of the LDC such as monocyte and basophil counts, and other commonly used indices such as red cell volume distribution width and platelet indices such as mean platelet volume and platelet distribution width are considered. The new parameters, evaluated from analytic and clinical viewpoints, are the available components of the extended differential count (hematopoietic progenitor cells, immature granulocytes, and erythroblasts), the immature reticulocyte fraction, the reticulocyte indices, the fragmented RBCs, and the immature platelet fraction.

Interobserver and intraobserver variation among experts in the diagnosis of thyroid follicular lesions with borderline nuclear features of papillary carcinoma.

Abstract: Distinguishing follicular variant of papillary carcinoma (FVPC) from follicular adenoma and follicular carcinoma can be difficult if nuclear features of papillary carcinoma are not well developed or only focally present. We assessed interobserver and intraobserver agreement among 6 thyroid experts by using 15 cases in which original pathologists suspected FVPC. There was unanimous expert agreement in diagnosing FVPC in only 2 cases (13%) and majority agreement in 6 cases (40%). Unanimous agreement on benign and malignant diagnoses was seen in 4 cases (27%) and majority agreement on malignancy in 8 cases (53%). Intraobserver agreement ranged from 17% to 100%. Histologic features considered most helpful in diagnosing FVPC were nuclear clearing, nuclear grooves, nuclear overlapping and crowding, nuclear membrane irregularity, and nuclear enlargement. This considerable interobserver and intraobserver variability in the diagnosis of FVPC seems to result from lack of agreement on the minimal criteria needed to diagnose FVPC, even among experts.

CSF multianalyte profile distinguishes Alzheimer and Parkinson diseases.

Abstract: The therapeutic imperative for Alzheimer disease (AD) and Parkinson disease (PD) calls for discovery and validation of biomarkers. Increased cerebrospinal fluid (CSF) τ and decreased amyloid (A) β 42 have been validated as biomarkers of AD. In contrast, there is no validated CSF biomarker for PD. We validated our proteomics-discovered multianalyte profile (MAP) in CSF from 95 control subjects, 48 patients with probable AD, and 40 patients with probable PD. An optimal 8-member MAP agreed with expert diagnosis for 90 control subjects (95%), 36 patients with probable AD (75%), and 38 patients with probable PD (95%). This MAP consisted of the following (in decreasing order of contribution): τ, brain-derived neurotrophic factor, interleukin 8, Aβ 42 , β 2 -microglobulin, vitamin D binding protein, apolipoprotein (apo) AII, and apoE. This first large-scale validation of a proteomicdiscovered MAP suggests a panel of 8 CSF proteins that are highly effective at identifying PD and moderately effective at identifying AD. Alzheimer disease (AD) and Parkinson disease (PD) are major public health problems. Key to the effort to obtain new therapeutics will be novel biomarkers to aid in diagnosis, identify subsets of patients, and objectively monitor progression and response to treatment. Several discovery proteomic studies of human cerebrospinal fluid (CSF) have been reported using relatively small numbers of patients with AD compared with control subjects without dementia. 1-6

Glypican 3 expression in human nonneoplastic, preneoplastic, and neoplastic tissues: a tissue microarray analysis of 4,387 tissue samples.

Abstract: Several studies have shown that glypican 3 (GPC3) could be a useful diagnostic marker for hepatocellular carcinoma (HCC) and for differentiating HCC from nonneoplastic and preneoplastic liver disease. To systematically investigate the epidemiology of GPC3 expression in the liver and in other organs and tissues, we used tissue microarray technology comprising 4,387 tissue samples from 139 tumor categories and 36 nonneoplastic and preneoplastic tissue types. The immunohistochemical expression of GPC3 was assessed semiquantitatively using a 10% cutoff score and was detected in 9.2% of nonneoplastic liver samples (11/119), 16% of preneoplastic nodular liver lesions (6/38), and 63.6% of HCCs (140/220), underlining the role of GPC3 in hepatocarcinogenesis. Furthermore, several other tumors revealed consistent expression of GPC3, including squamous cell carcinoma of the lung (27/50 [54%]), testicular nonseminomatous germ cell tumors (32/62 [52%]), and liposarcoma (15/29 [52%]).

A new Mycobacterium species causing diffuse lepromatous leprosy.

Abstract: Mycobacterium leprae causes leprosy. M leprae strains collected worldwide have been genetically clonal, which poorly explains the varying severity and clinical features of the disease. We discovered a new Mycobacterium species from 2 patients who died of diffuse lepromatous leprosy (DLL). The Mycobacterium was purified from heavily infected, freshly frozen autopsy liver tissue followed by DNA extraction in 1 case. Paraffin-embedded skin tissue was used for DNA extraction in another case. Six genes of the organism were amplified by polymerase chain reaction, sequenced on cloning or from amplicons, and analyzed. Significant genetic differences with M leprae were found, including a 2.1% divergence of the 16S ribosomal RNA (rRNA) gene, a highly conserved marker of bacterial evolution, and 6% to 14% mismatches among 5 less conserved genes. Phylogenetic analyses of the genes of 16S rRNA, rpoB, and hsp65 indicated that the 2 most related organisms evolved from a common ancestor that had branched from other mycobacteria. These results and the unique clinicopathologic features of DLL led us to propose Mycobacterium lepromatosis sp nov. This species may account for some of the clinical and geographic variability of leprosy. This finding may have implications for the research and diagnosis of leprosy.

Biochemical markers for prediction of chemotherapy-induced cardiotoxicity: systematic review of the literature and recommendations for use.

Abstract: Chemotherapy is a well-established therapeutic approach for several malignancies, but its clinical efficacy is often limited by its related cardiotoxicity, which leads to cardiomyopathy, possibly evolving into heart failure. To detect cardiac damage, the adopted diagnostic approach is the estimation of left ventricular ejection fraction by echocardiography. This approach shows low sensitivity toward early prediction of cardiomyopathy, when the possibilities of appropriate treatments could still improve the patient's outcome. Cardiac troponins, however, show high diagnostic efficacy as early as 3 months before the clinical onset of cardiomyopathy. The increase in their concentrations is correlated with disease severity and may predict the new onset of major cardiac events during follow-up. Negative troponin concentrations may identify patients with a very low risk of cardiomyopathy (negative predictive value, 99%). Concerning cardiac natriuretic peptides, definitive evidence in regard to a diagnostic or prognostic role in predicting chemotherapy-induced cardiomyopathy is still lacking.

High-sensitivity tandem mass spectrometry assay for serum estrone and estradiol.

Abstract: High-sensitivity measurement of serum estrogens is important in adult and pediatric endocrinology and oncology. We developed a high-sensitivity liquid chromatography-tandem mass spectrometry (LC-MS/MS) assay for simultaneous measurement of estrone (E1) and estradiol (E2). Aliquots of 200 muL of serum were spiked with internal standard, extracted, derivatized with dansyl chloride, and analyzed by LC-MS/MS using 2-dimensional chromatographic separation. Total imprecision for the method was less than 11%; the limit of quantitation was 1 pg/mL. Reference intervals were established with samples from more than 900 healthy postmenopausal women, men, girls, and boys. Concentrations of estrogens in children reached adult levels by Tanner stage 3. In men and postmenopausal women, the median concentrations of total estrogens (E1 + E2) were 39 and 22 pg/mL, and the median E2/E1 ratios were 0.98 and 0.55, respectively. The method requires a small sample volume and has adequate sensitivity and specificity for analyzing estrogens in samples from postmenopausal women, men, and children.

High-resolution melting analysis for rapid detection of KRAS, BRAF, and PIK3CA gene mutations in colorectal cancer.

Abstract: High-resolution melting analysis (HRMA) provides a valid approach to efficiently detect DNA genetic and somatic mutations. In this study, HRMA was used for the screening of 116 colorectal cancers (CRCs) to detect hot-spot mutations in the KRAS and BRAF oncogenes. Mutational hot spots on the PIK3CA gene, exons 9 and 20, were also screened. Direct sequencing was used to confirm and characterize HRMA results. HRMA revealed abnormal melting profiles in 65 CRCs (56.0%), 16 of them harboring mutations in 2 different genes simultaneously. The frequency of mutations was 17.2% for PIK3CA (11.2% in exon 9 and 6.0% in exon 20), 43.1% for KRAS exon 2, and 9.5% in exon 15 of the BRAF gene. We found a significant association between PIK3CA and KRAS mutations (P = .008), whereas KRAS and BRAF mutations were mutually exclusive (P = .001). This report describes a novel approach for the detection of PIK3CA somatic mutations by HRMA.

Endobronchial ultrasound-guided transbronchial fine-needle aspiration: The University of Minnesota experience, with emphasis on usefulness, adequacy assessment, and diagnostic difficulties

Abstract: Endobronchial ultrasound-guided transbronchial fine-needle aspiration (EBUS-TBNA) is a new technique that facilitates cytologic sampling of mediastinal lymph nodes. We describe our initial experience with this method, including adequacy assessment, impact on cytopathologists' work, and diagnostic pitfalls. There were 229 EBUS-TBNA samples obtained from 100 patients; a mean of 22 minutes was spent with an average of 3 passes performed and 6 slides prepared per site. Of 193 aspirates, 5 were categorized as atypical, 54 as positive, and 134 as negative for malignancy; 36 (15.7%) aspirates were nondiagnostic. We found EBUS-TBNA to have a high specificity (100%) and good sensitivity (86%) in our institution, in which a cytopathologist is available on-site to ensure sample adequacy. Most true-negative samples had moderate to abundant lymphocytes, confirming lymphocyte numbers as a marker of adequacy. For pathologists, this was a relatively time-consuming procedure. Recognizing bronchial contamination, especially with metaplastic or dysplastic cells, is important for avoiding diagnostic pitfalls.

HER2+ breast cancer: review of biologic relevance and optimal use of diagnostic tools.

Abstract: Clinical laboratory testing for human epidermal growth factor receptor 2 (HER2) status in newly diagnosed breast cancer is critically important for therapeutic decision making. Unlike most pathologic testing, which serves as an adjunct to establishing a diagnosis, the results of HER2 testing stand alone in determining which patients are likely to respond to trastuzumab, a monoclonal antibody against HER2. Given the significant clinical impact of the testing results on subsequent patient management, the accuracy, precision, and reproducibility of HER2 testing are critical. At present, several preanalytic factors, including the time from tissue removal to tissue fixation, are underappreciated as important variables that have the potential to negatively impact the consistency and reliability of HER2 testing. Rigorous quality control and standardization of the testing process, from the handling of tissue samples to interpretation and reporting of results, are essential for achieving accurate and reproducible assay results.

Epidermal growth factor receptor expression and gene copy number in conventional hepatocellular carcinoma.

Abstract: Epidermal growth factor receptor (EGFR) is frequently overexpressed in hepatocellular carcinoma, but its relationship with EGFR gene copy number has not been studied. This study examined EGFR expression and gene copy number in hepatocellular carcinoma and evaluated their relationship to clinicopathologic features in 76 tumors. Moderate to strong expression of EGFR was observed by immunohistochemical analysis in 50 (66%) of 76 hepatocellular carcinomas. Fluorescence in situ hybridization (FISH) showed extra EGFR gene copies in 17 (45%) of 38 tumors. This was accompanied by gains of chromosome 7, indicating that this was the result of balanced polysomy rather than gene amplification. There was no correlation between EGFR expression by immunohistochemical analysis and gene copy number by FISH. EGFR expression showed borderline association with cirrhosis but not with other clinicopathologic parameters examined. EGFR overexpression is present in a majority of hepatocellular carcinomas, suggesting a role for EGFR antagonists in therapy. The increased expression does not correlate with an increase in the EGFR gene copy number.

Analysis of SOX9 Expression in Colorectal Cancer

Abstract: Our purpose was to investigate the role of SOX9, a novel downstream molecule of β-catenin, in colorectal cancer. Expression of SOX9 and β-catenin was detected by immunostaining, quantitative realtime reverse transcription–polymerase chain reaction (Q-PCR), and Western blot in colorectal cancer. The correlation between SOX9 or β-catenin expression and clinicopathologic parameters was also analyzed. Immunostaining, Q-PCR, and Western blot consistently confirmed SOX9 up-regulation in colorectal cancer compared with normal mucosa (P < .05). Immunostaining showed more SOX9+ cells in the lower zone of colonic crypts than in the upper zone (P < .05). Cancers with strong SOX9 immunostaining were significantly associated with a lower 5-year overall survival (40% [17/43] vs low expression, 69% [66/95]; P <.01). The Cox proportional hazards model showed that strong SOX9 expression was an independent adverse prognosticator in colorectal cancer (P < .05). The detection of SOX9 expression might contribute to predicting clinical outcomes for patients with colorectal cancer. Colorectal cancer is the second most common malignancy worldwide. Its prognosis has not been improved significantly during the past 2 or 3 decades. The elucidation of the molecu

Decreased mortality associated with prompt Gram staining of blood cultures.

Abstract: Gram stains of positive blood cultures are the most important factor influencing appropriate therapy The sooner appropriate therapy is initiated, the better Therefore, it is reasonable to expect that the sooner Gram stains are performed, the better To determine the value of timely Gram stains and whether improvement in Gram stain turnaround time (TAT) is feasible, we compared data for matched pairs of patients with cultures processed promptly (<1 hour TAT) with data for patients with cultures not processed promptly (≥1 hour TAT) and then monitored TAT by control charting In 99 matched pairs, average difference in time to detection of positive blood cultures within a pair of patients was less than 01 hour For the less than 1 hour TAT group, the average TAT and crude mortality were 01 hour and 101%, respectively; for the 1 hour or longer TAT group, they were 33 hours and 192%, respectively (P < 0001 and P = 0389, respectively) After multifaceted efforts, we achieved significant improvement in the TAT for Gram stains The Institute of Medicine has recommended that medical practices become more patient-centered 1 Two notable accomplishments are in the care of myocardial infarction and stroke In patients with acute myocardial infarction, reduced door-to-balloon time for primary angioplasty decreases mortality 2 Likewise, in the setting of stroke, studies suggest that

Lymphocytic esophagitis: a chronic or recurring pattern of esophagitis resembling allergic contact dermatitis.

Abstract: Lymphocytic esophagitis (LE) is characterized by intraepithelial lymphocytes (IELs) and spongiosis, resembling contact dermatitis. LE has been defined as high numbers of IELs and no or rare granulocytes and was found in young patients and in association with Crohn disease (CD). We reviewed the medical records of 42 LE cases. Cases were divided into severe (IELs in interpapillary and peripapillary fields) and mild (IELs in peripapillary fields) LE. The control group included specimens from 34 consecutive esophageal biopsy cases. Mean ages were similar (LE, 44 years; control subjects, 43 years). CD was present in 5 LE cases (12%) and 1 control case, an insignificant difference. Of patients with LE, 14 (33%) had an allergy; 11 (26%), gastroesophageal reflux disease (GERD); 4 (10%), Helicobacter pylori gastritis; and 18 (43%), dysphagia. No differences were found in clinical features between LE and control cases, except GERD was less common in severe LE (6/30 [20%]) than in control cases (17 [50%]). No patient with LE had celiac disease. No medications were common among LE cases. Patients with LE are statistically no more likely than control subjects to have CD. We found no association between LE and any clinical condition or symptom. Based on sequential biopsies in 7 patients, LE seems to be a chronic disease.

Diagnostic role of circulating free plasma DNA detection in patients with localized prostate cancer.

Abstract: To analyze the potential diagnostic relevance of free plasma DNA (FPDNA), we enrolled 64 patients with localized prostate cancer (CaP). FPDNA was quantified by real-time polymerase chain reaction assessment of the HTERT gene in blood samples from 64 patients with CaP and 45 healthy males. Methylation of the GSTP1 gene was used to confirm the neoplastic origin of FPDNA in selected cases. The mean +/- SD levels of FPDNA were higher in patients with CaP (15.4 +/- 10.9 ng/mL) than in control subjects (5.5 +/- 3.5 ng/mL; P <.001). By using the best cutoff value, the sensitivity of the test was 80%, the specificity was 82%, the area under the receiver operating characteristic curve, 0.881. High FPDNA values were significantly associated with pathologic T3 stage (P = . 035). Methylation of the GSTP1 gene was found in 4 (25%) of 16 FPDNA samples and 15 (94%) of 16 tissue samples. Quantification of FPDNA discriminates between patients with CaP and healthy subjects and correlates with pathologic tumor stage. FPDNA is a candidate biomarker for early diagnosis and monitoring of CaP.

Postmortem genetic testing for conventional autopsy-negative sudden unexplained death: an evaluation of different DNA extraction protocols and the feasibility of mutational analysis from archival paraffin-embedded heart tissue.

Abstract: One third of autopsy-negative sudden unexplained deaths (SUDs) can be attributed to a cardiac channelopathy. Typically, paraffin-embedded tissue (PET) is the only source of DNA available for genetic analyses. We examined different DNA extraction procedures, involving 2 deparaffinization methods, 2 digestion methods, 4 laboratory-based purification methods, and 5 commercial kits. Mutational analysis involving 25 RYR2 exons was performed on PET DNA from 35 SUD cases to evaluate the feasibility of using PET DNA for genetic testing. With the best PET-DNA extraction method, an average of only two thirds of the region of interest could be evaluated. Although we initially identified 5 missense mutations in 5 of 35 SUD cases, repeated analysis failed to confirm these mutations. DNA from PET should be considered error prone and unreliable in comprehensive surveillance of SUD-associated genes. Given these shortcomings, the standard autopsy for SUD should include archiving EDTA-preserved blood or frozen tissue to facilitate postmortem genetic testing.

Incidence, sensitivity, and specificity of leukemia-associated phenotypes in acute myeloid leukemia using specific five-color multiparameter flow cytometry.

Abstract: We assessed the usefulness of 5-color multiparameter flow cytometry to detect leukemia-associated phenotypes (LAPs) in the bone marrow of patients with newly diagnosed acute myeloid leukemia (AML) and determined its usefulness for detection of minimal residual disease (MRD). Overall, 94% of patients (51/54) with AML had LAPs at diagnosis. The frequency of leukemic bone marrow/median frequency of LAPs in normal or regenerating bone marrow samples using maximum log difference statistics revealed that CD2, CD56, CD11b, CD7, and CD19 expression on AML blasts represented the most sensitive and reliable markers for detection of MRD. Serial dilutional experiments showed that the sensitivity level of immunophenotyping was between 10-4 and 10-5 and that the approach was highly reproducible. Immunophenotypic analysis using a CD45 gating strategy, 5-color staining, and an extensive panel of monoclonal antibodies allowed the identification of LAPs in 94% of AML cases, and these immunophenotypes can be used for MRD monitoring with a sensitivity limit of 10-4 to 10-5.

The effect of elevated fetal hemoglobin on hemoglobin A1c results: five common hemoglobin A1c methods compared with the IFCC reference method

Abstract: Hemoglobin A 1c (HbA 1c ) is an important indicator of risk for complications in patients with diabetes mellitus. Elevated fetal hemoglobin (HbF) levels have been reported to interfere with results of some HbA 1c methods, but it has generally been assumed that HbA 1c results from boronate-affinity methods are not affected by elevated HbF levels. None of the previous studies used the International Federation of Clinical Chemistry and Laboratory Medicine (IFCC) reference method as the comparative HbA 1c method. We, therefore, measured HbA 1c in samples with normal and elevated HbF levels by several common assay methods and compared the results with those of the IFCC reference method. HbF levels of more than 20% artificially lowered HbA 1c results from the Primus CLC 330/385 (Primus Diagnostics, Kansas City, MO), Siemens DCA2000 (Siemens Healthcare Diagnostics, Tarrytown, NY), and Tosoh 2.2+ (Tosoh Bioscience, South San Francisco, CA), but not the Bio-Rad Variant II (Bio-Rad Laboratories, Hercules, CA) and Tosoh G7. Physicians and laboratory professionals need to be aware of potential interference from elevated HbF levels that could affect HbA 1c results, including those from boronate-affinity methods.

Evaluation of the value of frozen tissue section used as "gold standard" for immunohistochemistry.

Abstract: To examine the use of acetone- or ethanol-fixed frozen tissue sections as the "gold standard" for immunohistochemical analysis, we evaluated frozen sections with various conditions of fixation and antigen retrieval (AR). Fresh human tissues were frozen in OCT. An adjacent tissue block was fixed in 10% neutral buffered formalin (NBF) and paraffin embedded (FFPE). Frozen sections were fixed by 6 protocols: acetone, ethanol, NBF (2 durations), and NBF + calcium chloride (2 durations). AR was used for all NBF-fixed sections. More than half of the antibodies (16/26 [62%]) showed immunohistochemical results indistinguishable between acetone- and NBF-fixed sections; 8 (31%) showed better immunohistochemical signals following NBF and AR; 2 gave better immunohistochemical results for acetone-fixed sections. Most cytoplasmic proteins (10/13) showed comparable immunohistochemical signals between acetone- and NBF-fixed sections. For nuclear proteins, NBF-fixed sections gave better immunohistochemical signals than did acetone-fixed sections. In most cases, NBF yielded stronger signals with less background and better morphology. The data do not support the use of acetone-fixed frozen tissue sections as the gold standard for immunohistochemical analysis. In evaluating new antibodies, a combination of acetone- and NBF-fixed frozen sections should be used, although in practice, FFPE tissue sections may serve as the standard for most antigens for immunohistochemical analysis.

Immunohistochemical characterization of nasal-type extranodal NK/T-cell lymphoma using a tissue microarray: an analysis of 84 cases.

Abstract: Nasal-type extranodal natural killer (NK)/T-cell lymphoma is an uncommon malignancy. By using a tissue microarray, we characterized 84 cases of extranodal NK/T-cell lymphoma with regard to expression of 18 immunohistochemical markers and the presence of Epstein-Barr virus (EBV) RNA. In our series, CD2 was positive in 69 (93%) of 74 cases, CD3 in 68 (84%) of 81, CD5 in 22 (27%) of 81, CD20 in 0 (0%) of 82, CD29 in 75 (91%) of 82, CD30 in 29 (35%) of 84, CD43 in 81 (96%) of 84, CD54 in 58 (72%) of 81, CD56 in 46 (58%) of 79, CD62L in 23 (28%) of 83, CD183 in 66 (80%) of 83, BCL2 in 33 (39%) of 84, cutaneous lymphocyte antigen in 21 (25%) of 84, granzyme B in 70 (83%) of 84, Ki-67 in 59 (71%) of 83, linker for activation of T cells in 60 (71%) of 84, perforin in 66 (86%) of 77, TIA1 in 76 (90%) of 84, and EBV in 73 (87%) of 84. Hierarchical cluster analysis separated primary cutaneous cases from cases manifesting in other sites based on lower expression of the cell adhesion molecule CD54.

Clinical outcomes of ABO-incompatible RBC transfusions.

Abstract: Factors that predict outcome after ABO-incompatible RBC transfusions are not well defined. We studied whether the volume of incompatible blood transfused would determine the signs and symptoms and survival outcome for ABO-incompatible RBC transfusions. We reviewed ABO-incompatible RBC transfusions from our institutions and our consultations for 35 years and from a survey of America's Blood Centers' members regarding causes, volume, signs, symptoms, and outcomes of ABO-incompatible RBC transfusions in their service areas from 1995 through 2005. All ABO-incompatible transfusions were due to error; 26 (62%) of 42 occurred at the patient's bedside. Of 36 patients who received more than 50 mL of incompatible blood, 23 (64%) manifested signs or symptoms related to the incompatible transfusion, and 6 (17)% died. Only 3 (25%) of 12 patients who received 50 mL or less of incompatible blood had associated signs or symptoms, and none died. Hypotension, hemoglobinuria, and/or hemoglobinemia were the most frequent findings in survivors and patients who died.ABO-incompatible RBC transfusion does not inevitably mean death or even occurrence of symptoms. Prompt recognition and discontinuation of the transfusion are critical because transfusing less ABO-incompatible blood may minimize signs and symptoms and may prevent death.

Expression of glypican 3 in ovarian and extragonadal germ cell tumors.

Abstract: Germ cell tumors (GCTs), rare malignancies that occur in a wide range of locations and display variable histologic patterns, may pose diagnostic challenges. Glypican 3 (GPC3), a membrane-bound heparan sulfate proteoglycan, has been shown to be a novel diagnostic marker in testicular GCT. However, GPC3 expression in ovarian and extragonadal GCT has not been reported. We evaluated GPC3 immunoreactivity in GCTs from 63 patients (57 children and 6 adults), including 14 ovarian and 20 extragonadal primary GCTs and 8 metastases along with 21 primary testicular GCTs for comparison. All 33 yolk sac tumors (YSTs) and both choriocarcinomas were immunoreactive for GPC3. In contrast, a minority of immature (4/10) and mature (4/35) teratomas were positive. No positivity was seen in 6 embryonal carcinomas or 5 germinomas. GPC3 is differentially expressed in ovarian and extragonadal GCTs, with expression predominantly observed in YSTs and choriocarcinoma.

Comparison of Sebia Capillarys Capillary Electrophoresis With the Primus High-Pressure Liquid Chromatography in the Evaluation of Hemoglobinopathies

Abstract: The Sebia Capillarys (capillary zone electrophoresis [CE]) and the Primus Resolution high-pressure liquid chromatography (HPLC) were used to prospectively evaluate 297 samples for hemoglobinopathies. Hemoglobin (Hb) A levels were similar on both techniques (mean, 96.2% and SD, 5.7% by CE; mean, 96.8% and SD, 5.5% by HPLC), but HbA2 levels were higher by CE (mean, 2.8%; SD, 0.8%) than by HPLC (mean, 2.3%; SD, 0.8%). HbS had higher values by CE (mean, 40.6%; SD, 18.9%) than by HPLC (mean, 38.4%; SD, 18.9%). In cases with Hg S, HbA2 levels were greater by HPLC (mean, 4.0%; SD, 1.0%) than by CE (mean, 3.1%; SD, 0.8%). HbA2 was occasionally not separated sufficiently from HbC for measurement by CE, but did separate from HbE by CE. Both methods identified HbS, HbC, HbE, HbS, and HbC together, HbA2&prime, HbD-Los Angeles, HbF variant, HbG-Philadelphia, HbS-G Philadelphia, and Hb Lepore.

Prognostic Significance of Intratumoral and Peritumoral Lymphatic Density and Blood Vessel Density in Invasive Breast Carcinomas

Abstract: We studied tumor lymphatic and vascular densities and lymphovascular invasion (LVI) as prognostic markers in 48 cases of invasive breast cancer treated with partial or total mastectomy and lymph node dissection. All cases were immunostained with D2-40 and CD31. Positively stained microvessels were counted in densely vascular/lymphatic foci (hot spots) at x400. The mean+/-SD peritumoral lymphatic microvessel density (LMD) was significantly higher than intratumoral LMD (9+/-7 vs 4+/-6; P< .01). There was a positive correlation of D2-40 LMD (peritumoral and intratumoral) and CD31 microvessel density counts with lymph node metastasis (r=0.35, 0.5, and 0.38), nuclear grade (r=0.36, 0.28, and 0.3), and stage (r=0.42, 0.56, and 0.49), respectively. Peritumoral and intratumoral D2-40 LMD correlated significantly with the presence of angiolymphatic invasion (detected by D2-40; r=0.54 and 0.54, respectively). D2-40 detected more LVI than H&E- and CD31-detectable vascular invasion (18/48, 5/48, 11/48, respectively). Increased D2-40 detected LVI, and high CD31 microvessel counts showed significant adverse effect on survival status.

Parameters of thromboelastography in healthy newborns

Abstract: Thromboelastography (TEG) aids in monitoring a patient’s global hemostatic system by measuring the rate of clot formation, clot strength, and stability. The usefulness of TEG in pediatric settings, especially with neonates, is limited owing to a lack of neonatal reference values. In this study, neonatal TEG reference intervals were developed and results correlated with other coagulation test parameters. Samples were from women who delivered a neonate after at least 34 weeks of gestation in normal pregnancies. From the recovered placenta, cord blood from the umbilical vein or artery was collected within 30 minutes after delivery and tested. Neonatal TEG reaction time (time clot formation begins), clot firmness (shear elastic modulus strength), and platelet function analysis closure times were significantly lower than those in adult ranges (P < .001). When compared with the values for children, TEG reaction time, angle, coagulation index, clot firmness value, and clot kinetics (time from clot formation to time amplitude reaches 20 mm) were significantly different (P < .001) among neonates. TEG can be used to interpret the data for newborns by using reference values obtained in the present study. All patients with a bleeding or thrombotic history need

Performance of commercial platforms for rapid genotyping of polymorphisms affecting warfarin dose.

Abstract: Initiation of warfarin therapy is associated with bleeding owing to its narrow therapeutic window and unpredictable therapeutic dose. Pharmacogenetic-based dosing algorithms can improve accuracy of initial warfarin dosing but require rapid genotyping for cytochrome P-450 2C9 (CYP2C9) *2 and *3 single nucleotide polymorphisms (SNPs) and a vitamin K epoxide reductase (VKORC1) SNP. We evaluated 4 commercial systems: INFINITI analyzer (AutoGenomics, Carlsbad, CA), Invader assay (Third Wave Technologies, Madison, WI), Tag-It Mutation Detection assay (Luminex Molecular Diagnostics, formerly Tm Bioscience, Toronto, Canada), and Pyrosequencing (Biotage, Uppsala, Sweden). We genotyped 112 DNA samples and resolved any discrepancies with bidirectional sequencing. The INFINITI analyzer was 100% accurate for all SNPs and required 8 hours. Invader and Tag-It were 100% accurate for CYP2C9 SNPs, 99% accurate for VKORC1 -1639/3673 SNP, and required 3 hours and 8 hours, respectively. Pyrosequencing was 99% accurate for CYP2C9 *2, 100% accurate for CYP2C9 *3, and 100% accurate for VKORC1 and required 4 hours. Current commercial platforms provide accurate and rapid genotypes for pharmacogenetic dosing during initiation of warfarin therapy.

Discrepancies between clinical and autopsy diagnoses: a comparison of university, community, and private autopsy practices.

Abstract: Although it is known that autopsies often disclose unexpected findings, few studies have been published that address the effect of institutional setting, selection bias, and length of hospitalization. Records of medical autopsies from 3 institutional settings were studied for discrepancies between clinical and autopsy findings. The settings were university hospital (n = 85); community hospital (n = 146); and private autopsy (n = 60), which were referred from various community hospitals and paid for by family members. The same prosector performed the autopsies in the community and private settings. The overall rate of major discrepancy that involved the cause of death was 17.2%. Factors that increased the likelihood of missed diagnoses were private setting (P = .0005), community setting (P = .02), and short hospital stay before death (P = .02). Additional major findings were present in 28.5% of autopsies. Length of hospital stay before death, institution, and selection bias all affect the rate of major unexpected findings in hospital-based autopsies.

In situ PCR for Mycobacterium tuberculosis in endoscopic mucosal biopsy specimens of intestinal tuberculosis and Crohn disease.

Abstract: Tuberculosis and Crohn disease are granulomatous disorders affecting the intestinal tract with similar clinical manifestations and pathologic features. We evaluated the use of in situ polymerase chain reaction (PCR) using Mycobacterium tuberculosis complex-specific primers for IS 6110 to differentiate these 2 disorders in archival mucosal biopsy specimens. In situ PCR was positive in 6 of 20 tuberculosis biopsy specimens and 1 of 20 Crohn disease biopsy specimens. Staining was localized to a site of granulomatous inflammation in 3 of the tuberculosis specimens and in the Crohn disease specimen. In the other tuberculosis biopsy specimens, positive staining was localized to inflammatory granulation tissue and to a focus of intact mucosa without granulomatous inflammation. The presence of M tuberculosis DNA in Crohn disease could be due to coexisting latent tuberculosis or indicate a role for these bacteria in triggering an abnormal immune response. Therefore, in situ PCR is potentially useful to differentiate intestinal tuberculosis from Crohn disease, if the sensitivity is improved.

Value of Glypican 3 Immunostaining in the Diagnosis of Hepatocellular Carcinoma on Needle Biopsy

Abstract: The diagnostic value of glypican 3 (GPC3) immunostaining on needle biopsy specimens has not been well assessed. In this study, 120 liver needle biopsy specimens, including 46 from cirrhotic livers and 74 hepatocellular carcinomas (HCCs), were immunohistochemically examined for expression of GPC3. The results showed strong cytoplasmic and membranous staining in 36 HCCs (49%), among which 20 cases (56%) showed diffuse immunoreactivity. None of the 46 cirrhotic livers exhibited positive GPC3 immunostaining. The nonneoplastic liver tissues (cirrhotic or noncirrhotic) that were present in the majority of the HCC cases were also completely negative for GPC3 expression. These data demonstrate that GPC3 is a reliable immunohistochemical marker for the diagnosis of HCC on needle biopsy specimens when positive. However, the detection rate in our series seems lower than that reported in studies using resection specimens as the study materials. Our findings emphasize that GPC3 immunoreactivity can be focal and that negative staining should not be viewed as evidence to exclude the diagnosis of HCC in challenging needle biopsy specimens.

HER2 overexpression and amplification in urothelial carcinoma of the bladder is associated with MYC coamplification in a subset of cases

Abstract: We examined 53 invasive high-grade urothelial carcinomas (UCCs) and 42 paired lymph node metastases to determine frequency of HER2 overexpression, HER2/MYC coamplification, and association between HER2 and MYC status and clinicopathologic features. HER2 overexpression occurred in 19 UCCs (36%) and 14 metastases (30%), with an 88% concordance rate between UCCs and matched metastases. HER2 amplification occurred in 5 (10%) of 50 UCCs and 4 (11%) of 36 metastases, with a 100% concordance rate; MYC amplification occurred in 7 (18%) of 40 UCCs and 4 (13%) of 32 metastases, with a concordance rate of 50% between UCCs and metastases. Of 7 cases demonstrating HER2 amplification, MYC was coamplified in 4 (57%; P = .01), and coamplification was associated in all cases with metastasis and advanced local disease (pT4). Coamplification of HER2 and MYC occurs in a subset of patients with metastatic UCC. HER2 overexpression and amplification in metastatic lesions suggest that HER2-targeted therapy may be valuable for patients undergoing treatment for metastatic UCC, for which current therapy is limited. Further studies into the role of MYC coamplification in this population are needed to determine impacts on treatment with HER2-targeted therapy.