Best Practice in Diagnostic Immunohistochemistry: Prostate Carcinoma and Its Mimics in Needle Core Biopsies

2008 ◽  
Vol 132 (9) ◽  
pp. 1388-1396
Author(s):  
Gladell P. Paner ◽  
Daniel J. Luthringer ◽  
Mahul B. Amin
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Basal Cell ◽  
Prostate Carcinoma ◽  
Best Practice ◽  
English Language ◽  
Specific Antigen ◽  
Specific Marker ◽  
Nephrogenic Adenoma ◽  
Cytokeratin 5

Abstract Context.—The unrelenting challenge encountered when differentiating limited-volume prostate carcinoma and sometimes subtle variants from its many morphologic mimics has increased the use of ancillary immunohistochemistry in routine prostate needle biopsies. The availability of prostate cancer–associated and basal cell–associated markers has been an invaluable addition to diagnostic surgical pathology. Objective.—To review commonly used immunohistochemical stains, including innovative combinations, for confirmation or differential diagnosis of prostate carcinoma, and to propose appropriately constructed panels using morphologic patterns in prostate needle biopsies. Data Sources.—These best practices are based on our experience with routine and consultative case sign-outs and on a review of the published English-language literature from 1987 through 2008. Conclusions.—Basal cell–associated markers p63, high-molecular-weight cytokeratin 34βE12, cytokeratin 5/6 or a cocktail containing p63 and high-molecular-weight cytokeratin 34βE12 or cytokeratin 5/6 and prostate carcinoma–specific marker α-methylacyl coenzyme A (coA) racemase alone or in combination are useful adjuncts in confirming prostatic carcinoma that either lacks diagnostic, qualitative or quantitative features or that has an unusual morphologic pattern (eg, atrophic, pseudohyperplastic) or is in the setting of prior treatment. The combination of α-methylacyl coA racemase positivity with negative staining for basal cell–associated markers supports a malignant diagnosis in the appropriate morphologic context. Dual chromogen basal cell– associated markers (p63 [nuclear] and high-molecular-weight cytokeratin 34βE12/cytokeratin 5/6 [cytoplasmic]) and α-methylacyl coA racemase in an antibody cocktail provide greater sensitivity for the basal cell layer, easing evaluation and minimizing loss of representation of the focal area interest because the staining is performed on one slide. In the posttreatment setting, pancytokeratin facilitates detection of subtle-treated cancer cells. Prostate-specific antigen and prostatic acid phosphatase markers are helpful in excluding secondary malignancies involving the prostate, such as urothelial carcinoma, and occasionally in excluding nonprostatic benign mimickers, such as nephrogenic adenoma, mesonephric gland hyperplasia, and Cowper glands. There is no role for ordering immunohistochemistry prospectively in all cases of prostatic needle biopsies.

scholarly journals A Strain-Specific Antigen in Japanese Helicobacter pylori Recognized in Sera of Japanese Children

2005 ◽  
Vol 12 (11) ◽  
pp. 1280-1284 ◽  
Author(s):  
Masumi Okuda ◽  
Toshiro Sugiyama ◽  
Kenichi Fukunaga ◽  
Masaru Kondou ◽  
Eikichi Miyashiro ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Helicobacter Pylori ◽  
High Molecular Weight ◽  
Early Stage ◽  
False Negative ◽  
Specific Antigen ◽  
Host Immune System ◽  
Japanese Strain ◽  
Asymptomatic Children ◽  
Associated Proteins

ABSTRACT An enzyme immuno assay (EIA) test based on Japanese strain-derived high-molecular-weight cell-associated proteins (JHM-CAP) was evaluated by comparing with a previously developed EIA test based on a U.S. strain-derived high-molecular-weight cell-associated proteins (HM-CAP). Serum samples of 131 Japanese asymptomatic children (mean age, 5.5 years; range, 0 to 21 years) were tested that include 43 positive and 88 negative children as judged by Helicobacter pylori stool antigen test (HpSA test). Both tests showed comparable and reliable specificities, but the sensitivity of JHM-CAP EIA, at 93.0%, was much higher than that of HM-CAP EIA, at 67.4%. More false-negative results of HM-CAP were obtained in children under 10 years of age. Immunoblot analysis revealed that the JHM-CAP but not the HM-CAP preparation had a 100-kDa antigen recognized by JHM-CAP positive sera. It was concluded that JHM-CAP EIA is highly accurate for the serodiagnosis of H. pylori infection in Japanese young children and that the high sensitivity of JHM-CAP EIA in contrast to HM-CAP EIA is due to the presence of a 100-kDa antigen in Japanese strains that may be recognized by the host immune system at an early stage of infection.

scholarly journals Prostate-Specific Antigen, High-Molecular-Weight Cytokeratin (Clone 34βE12), and/or p63

2006 ◽  
Vol 125 (5) ◽  
pp. 675-681 ◽  
Author(s):  
Lakshmi P. Kunju ◽  
Rohit Mehra ◽  
Matthew Snyder ◽  
Rajal B. Shah
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Prostate Specific Antigen ◽  
Specific Antigen

scholarly journals Dextranol: A better lyoprotectant

2018 ◽  
Author(s):  
Bryan Joshua Jones ◽  
Advitiya Mahajan ◽  
Alptekin Aksan
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Room Temperature ◽  
Elevated Temperatures ◽  
Serum Proteins ◽  
Specific Antigen ◽  
Clinical Samples ◽  
Serum Samples ◽  
Protein Molecules ◽  
Anomeric Carbon

Dextranol, a reduced dextran, prevents damage to stored dry protein samples that unmodified dextran would otherwise cause. Lyoprotectants like the polysaccharide dextran are critical for preserving dried protein samples by forming rigid a glass that protects entrapped protein molecules. Stably dried proteins are important for maintaining critical information in clinical samples like blood serum. However, we found that dextran reacts with serum proteins during storage, producing high-molecular weight Amadori-product conjugates. These conjugates appeared in a matter of days or weeks when stored at elevated temperatures (37° or 45°C), but also appeared on a timescale of months when stored at room temperature. We synthesized a less reactive dextranol by reducing dextran’s anomeric carbon from an aldehyde to an alcohol. Serum samples dried in a dextranol-based matrix protected the serum proteins from forming high-molecular weight conjugates. The levels of four cancer-related serum biomarkers (prostate specific antigen, neuropiln-1, osteopontin, and metalloproteinase 7) decreased, as measured by immunoassay, when serum samples were stored for one to two weeks in dextran-based matrix. Switching to a dextran-based lyoprotection matrix slightly reduced the damage to osteopontin and completely stopped any detectable damage during storage in the other three biomarkers when for a period of two weeks at 45°C. Dextranol offers a small and easy modification to dextran that significantly improves the molecule’s function as a lyoprotectant by eliminating the potential for damaging protein-polysacharide conjugation.

scholarly journals The extracellular domain of CD45 controls association with the CD4-T cell receptor complex and the response to antigen-specific stimulation.

1996 ◽  
Vol 183 (1) ◽  
pp. 249-259 ◽  
Author(s):  
D Leitenberg ◽  
T J Novak ◽  
D Farber ◽  
B R Smith ◽  
K Bottomly
Keyword(s):  
Molecular Weight ◽  
T Cell ◽  
High Molecular Weight ◽  
T Cell Receptor ◽  
Tyrosine Phosphatase ◽  
Specific Antigen ◽  
Cell Receptor ◽  
Low Molecular Weight ◽  
Cell Surface Antigens ◽  
Cd45 Isoforms

The CD45 tyrosine phosphatase plays an important role in regulating T lymphocyte activation, but the function of the different isoforms of CD45 is not known. T cell transfectants have been prepared that express individual CD45 isoforms in cells with a well-defined T cell receptor (TCR) from the D10 T helper 2 clone. We find that cells bearing low molecular weight CD45 isoforms are far more efficient in responding to stimulation with peptide and antigen-presenting cells compared with cells bearing high molecular weight CD45 isoforms. One hypothesis for the preferential activation of cells that express low molecular weight CD45 isoforms is that they interact with other cell surface antigens important in TCR signaling, altering their phosphorylation status and affecting the character of the signal transduction pathway. In this report, using cells expressing single isoforms, we demonstrate that low molecular weight isoforms of CD45 preferentially associate with CD4 and the TCR complex compared with high molecular weight isoforms. The molecular basis for this interaction was further examined using a glycosyl phosphatidyl inositol (GPI)-linked form of CD45Null (lacking tyrosine phosphatase domains), which preferentially associated with CD4 compared with GPI-linked CD45ABC, and cytoplasmic tail mutants of CD4, which retained the ability to coassociate. Using this panel of transfectants, it is clear that the interaction between CD4 and CD45 does not require the cytoplasmic domains of CD45, but is dependent on the specific external domain of the various isoforms: low molecular weight species were more likely to associate with the CD4-TCR complex than the higher molecular weight isoforms, and their ability to coassociate correlated with the magnitude of the response to specific antigen.

Diagnostic Uncertainty Expressed in Prostate Needle Biopsies

1999 ◽  
Vol 123 (8) ◽  
pp. 687-692 ◽  
Author(s):  
David A. Novis ◽  
Richard J. Zarbo ◽  
Paul A. Valenstein
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Prostate Specific Antigen ◽  
Needle Biopsy ◽  
Specific Antigen ◽  
The United States ◽  
Immunoperoxidase Staining ◽  
Improvement Program ◽  
Prostate Needle Biopsy ◽  
Diagnostic Uncertainty

Abstract Objective.—To determine the rate of diagnostic uncertainty in rendering diagnoses on prostate needle biopsies and to examine pathology practice variables that influence that rate. Design.—Anatomic pathology departments participating in the College of American Pathologists Q-Probes laboratory quality improvement program retrospectively reviewed their last 50 consecutive prostate needle biopsy diagnoses. For each diagnosis, participants provided information concerning patients' prostate-specific antigen levels; number, locations, and laterality of biopsy specimens; number of tissue levels examined; performance of high-molecular-weight cytokeratin immunoperoxidase staining; and acquisition of consultations from general pathologists or experts in prostate pathology. Characteristics of pathology practices included yearly surgical and prostate needle biopsy caseloads, number of pathologists rendering biopsy diagnoses, use of standard descriptive checklists, access to patients' prostate-specific antigen and digital rectal examination results, percentages of prostate needle biopsies routinely submitted for internal consultations, and presence of departmental experts in prostate pathology. Setting and Participants.—Three hundred thirty-two public and private institutions located in the United States (n = 318), Canada (n = 6), Australia (n = 5), United Kingdom (n = 2), and Guam (n = 1). Main Outcome Measure.—The rate of diagnostic uncertainty in prostate needle biopsy diagnoses. Results.—Participants submitted diagnoses on a total of 15 753 prostate needle biopsy cases, of which 33.4% were adenocarcinoma; 55.5% were benign; 3.9% were carcinoma in situ, prostatic intraepithelial neoplasia, or both; and 7.1% were diagnostically uncertain. The median rate of diagnostic uncertainty was 6%, ranging from 0 at the 10th percentile to 14% at the 90th percentile of all participating laboratories. Performing high-molecular-weight cytokeratin immunoperoxidase staining resolved diagnostic uncertainty in 68% of cases in which it was performed, and obtaining intradepartmental and extradepartmental consultations resolved diagnostic uncertainty in 70% to 87% of cases for which they were obtained. Knowledge of patients' prostate-specific antigen results and examining multiple biopsy cores had marginal effects on the rate of uncertainty. Thoroughness of prostate gland sampling and examination of multiple tissue block levels were not associated with the aggregate rate of diagnostic uncertainty. We found no particular pathology departmental practices or institutional demographic characteristics associated with institutional rates of diagnostic uncertainty. Conclusions.—Use of high-molecular-weight cytokeratin immunoperoxidase staining and obtaining intradepartmental and extradepartmental consultations may be effective in reducing diagnostic uncertainty in prostate biopsies.

Progressive Exposure of E-Neoantigen Associated with Degradation of Crosslinked Fibrin by Plasmin In Vitro

1984 ◽  
Vol 52 (03) ◽  
pp. 315-320
Author(s):  
Mojca Stegnar ◽  
James P Chen
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Conformational Changes ◽  
Protein Fraction ◽  
Specific Marker ◽  
Double Antibody ◽  
Terminal Fragment ◽  
Partial Degradation ◽  
Related Proteins

SummaryFragment E-neoantigen (Eneo) is a specific marker of structural and conformational changes associated with the degradation of fibrinogen and its related proteins. In this study Eneo expression was followed during the plasmin degradation of crosslinked (XL) fibrin in vitro, utilizing double antibody radioimmunoassay. Eneo was progressively exposed as degradation proceeded and its expression was associated with the liberation of high molecular weight fragments from XL-fibrin, their degradation into fragments DD and E, and the partial degradation of fragment E itself. An isolated protein fraction containing early high molecular weight fragments (MW >360000 d) and a fraction containing fragments YY and DY expressed approximately 170 times and 15 times less Eneo per protein respectively compared to the fragment E standard. XL- fibrin fragment E isolated from 1-hr plasmin digest expressed approximately 8 times less Eneo than fragment E isolated from 24-hr digest, indicating increased exposure of Eneo during fragment E degradation. Eneo expression of this terminal fragment E was comparable to fibrinogen fragment E. As expected, fragment DD had no Eneo immunoreactivity.

scholarly journals Plastics and the microbiome: impacts and solutions

2021 ◽  
Vol 16 (1) ◽  
Author(s):  
G. Lear ◽  
J. M. Kingsbury ◽  
S. Franchini ◽  
V. Gambarini ◽  
S. D. M. Maday ◽  
...  
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Best Practice ◽  
Structural Integrity ◽  
Degradation Products ◽  
Abiotic Factors ◽  
Carbon Sources ◽  
Polymer Degradation ◽  
Microbial Biodegradation ◽  
Negative Impacts

AbstractGlobal plastic production has increased exponentially since manufacturing commenced in the 1950’s, including polymer types infused with diverse additives and fillers. While the negative impacts of plastics are widely reported, particularly on marine vertebrates, impacts on microbial life remain poorly understood. Plastics impact microbiomes directly, exerting toxic effects, providing supplemental carbon sources and acting as rafts for microbial colonisation and dispersal. Indirect consequences include increased environmental shading, altered compositions of host communities and disruption of host organism or community health, hormone balances and immune responses. The isolation and application of plastic-degrading microbes are of substantial interest yet little evidence supports the microbial biodegradation of most high molecular weight synthetic polymers. Over 400 microbial species have been presumptively identified as capable of plastic degradation, but evidence for the degradation of highly prevalent polymers including polypropylene, nylon, polystyrene and polyvinyl chloride must be treated with caution; most studies fail to differentiate losses caused by the leaching or degradation of polymer monomers, additives or fillers. Even where polymer degradation is demonstrated, such as for polyethylene terephthalate, the ability of microorganisms to degrade more highly crystalline forms of the polymer used in commercial plastics appears limited. Microbiomes frequently work in conjunction with abiotic factors such as heat and light to impact the structural integrity of polymers and accessibility to enzymatic attack. Consequently, there remains much scope for extremophile microbiomes to be explored as a source of plastic-degrading enzymes and microorganisms. We propose a best-practice workflow for isolating and reporting plastic-degrading taxa from diverse environmental microbiomes, which should include multiple lines of evidence supporting changes in polymer structure, mass loss, and detection of presumed degradation products, along with confirmation of microbial strains and enzymes (and their associated genes) responsible for high molecular weight plastic polymer degradation. Such approaches are necessary for enzymatic degraders of high molecular weight plastic polymers to be differentiated from organisms only capable of degrading the more labile carbon within predominantly amorphous plastics, plastic monomers, additives or fillers.

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