Endogenous avidin biotin activity (EABA) in thyroid pathology: immunohistochemical study

Nikiel, Barbara; Chekan, Mykola; Jarząb, Michał; Lange, Dariusz

doi:10.1186/1756-6614-2-5

Cited by 11 publications

(7 citation statements)

References 18 publications

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“…The possible explanation for such discrepancy comes from the knowledge that Hürthle cells are rich in endogenous biotin, which may provide false positive results in Hürthle cell tumors if biotin-based detection systems are used. Since the presence of biotin has been also documented in follicular adenomas, follicular carcinomas, and papillary carcinoma [15,16,22], the use of biotin-free systems appears to be extremely relevant when immunohistochemistry is performed in thyroid lesions. The artifacts due to the presence of endogenous biotin are known to be markedly enhanced when heat-induced procedures for antigen retrieval are carried out during immunohistochemical protocols [33].…”

Section: Discussionmentioning

confidence: 99%

“…For the immunohistochemical investigation of thyroid diseases, it is very important to choose an optimal visualization method and an appropriate quality control, as it was shown that the presence of endogenous biotin in benign and malignant follicular cells may determine false positive immunohistochemical results [15,16]. Widely used and highly sensitive immunohistochemical methods, such as the labeled streptavidin biotin (LSAB) method, based on the high affinity of streptavidin and biotin, may lead to non-specific reactions in paraffinembedded material, caused by the interaction between streptavidin and endogenous biotin.…”

Section: Introductionmentioning

confidence: 99%

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Diagnostic value of neutrophil gelatinase-associated lipocalin (NGAL) immunoexpression in follicular-patterned lesions of the thyroid gland

et al. 2012

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Neutrophil gelatinase-associated lipocalin (NGAL) is a protein which participates in iron trafficking and which is involved in cancerogenesis and cancer progression. Since its over-expression has been documented in thyroid malignancies in comparison to thyroid normal gland, in the present study, we aimed to determine whether the evaluation of NGAL immunoexpression may be of help in the differential diagnosis of follicular-patterned thyroid lesions. Our additional aim was to test the possible interference of endogenous biotin on the immunohistochemical findings. Thus, all the immunohistochemical procedures, carried out with labeled streptavidin biotin method, were doubly performed, with or without the preliminary inhibition of endogenous biotin. No NGAL staining was found in the normal thyroid gland nor in the nodular colloid goiters or in Hashimoto's thyroiditis. NGAL expression appeared to be significantly more frequent in the malignant tumors in comparison to benign ones (P < 0.000001). Even more, NGAL expression appeared to be specific (specificity 93%) for carcinoma and represented a sensitive method (sensitivity 84%), with high negative (88%) and positive (94%) predictive values, as well as high diagnostic accuracy (88%), in the identification of follicular-patterned thyroid malignant tumors. The specificity, positive predictive value and diagnostic accuracy lowered when biotin was not preliminary inhibited, due to the presence of false positives among benign Hürthle cell tumors. In conclusion, the immunohistochemical detection of NGAL may be helpful in the differential diagnosis between malignant and benign follicular-patterned lesions of the thyroid. The use of biotin free system or the preliminary biotin inhibition is warranted for the detection of NGAL in thyroid samples, especially when dealing with Hürthle cell tumors.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Diagnostic value of neutrophil gelatinase-associated lipocalin (NGAL) immunoexpression in follicular-patterned lesions of the thyroid gland

et al. 2012

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“…138 EABA is accentuated by HIER but also develops in tissues subjected to other types of AR. 56,187,239,257,275 EABA is typically in the cytoplasm but has been reported in the nucleus. 250,257,342,428 Commercially available EABA blocking reagents (pure avidin and biotin solutions) are expensive, so many laboratories use ''homemade'' solutions with egg white as a source of avidin and 5% powdered milk as a source of biotin.…”

Section: Detection Systems: the Sensitivity Issuementioning

confidence: 99%

“…56,187,239,257,275 EABA is typically in the cytoplasm but has been reported in the nucleus. 250,257,342,428 Commercially available EABA blocking reagents (pure avidin and biotin solutions) are expensive, so many laboratories use ''homemade'' solutions with egg white as a source of avidin and 5% powdered milk as a source of biotin. 12,84,175,239,285,418 Non-Avidin-Biotin Methods Peroxidase-Antiperoxidase (PAP) Method.…”

Section: Detection Systems: the Sensitivity Issuementioning

confidence: 99%

When Tissue Antigens and Antibodies Get Along

2013

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Once focused mainly on the characterization of neoplasms, immunohistochemistry (IHC) today is used in the investigation of a broad range of disease processes with applications in diagnosis, prognostication, therapeutic decisions to tailor treatment to an individual patient, and investigations into the pathogenesis of disease. This review addresses the technical aspects of immunohistochemistry (and, to a lesser extent, immunocytochemistry) with attention to the antigen-antibody reaction, optimal fixation techniques, tissue processing considerations, antigen retrieval methods, detection systems, selection and use of an autostainer, standardization and validation of IHC tests, preparation of proper tissue and reagent controls, tissue microarrays and other high-throughput systems, quality assurance/quality control measures, interpretation of the IHC reaction, and reporting of results. It is now more important than ever, with these sophisticated applications, to standardize the entire IHC process from tissue collection through interpretation and reporting to minimize variability among laboratories and to facilitate quantification and interlaboratory comparison of IHC results.

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“…There are three possible other reasons that could account for nuclear staining for biotin. It may be possible that the nuclear positive gold staining could be caused by nonspecific staining as a result of secondary labeling of biotin in newly replicated DNA [38, 39], or endogenous biotin, which has widely been taken up in the cell nuclei [40], or biotin could have come from biodegradation of the anti-DMPO probe. As the control IgG contrast agent sample had little staining for biotin, we surmise that the biotin detected in the cell nuclei from the glioma sample administered the anti-DMPO probe was from biodegradation of the probe which had accumulated in the glioma cells of untreated tumors.…”

Section: Discussionmentioning

confidence: 99%

OKN-007 decreases free radical levels in a preclinical F98 rat glioma model

Souza

Smith

Atolagbe

et al. 2015

Free Radical Biology and Medicine

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Free radicals are associated with glioma tumors. Here, we report on the ability of an anticancer nitrone compound, OKN-007 [Oklahoma Nitrone 007; a disulfonyl derivative of α-phenyl-tert-butyl nitrone (PBN)] to decrease free radical levels in F98 rat gliomas using combined molecular magnetic resonance imaging (mMRI) and immunospin-trapping (IST) methodologies. Free radicals are trapped with the spin-trapping agent, 5,5-dimethyl-1-pyrroline N-oxide (DMPO), to form DMPO macromolecule radical adducts, and then further tagged by immunospin trapping by an antibody against DMPO adducts. In this study, we combined mMRI with a biotin–Gd-DTPA–albumin-based contrast agent for signal detection with the specificity of an antibody for DMPO nitrone adducts (anti-DMPO probe), to detect in vivo free radicals in OKN-007-treated rat F98 gliomas. OKN-007 was found to significantly decrease (P < 0.05) free radical levels detected with an anti-DMPO probe in treated animals compared to untreated rats. Immunoelectron microscopy was used with gold-labeled antibiotin to detect the anti-DMPO probe within the plasma membrane of F98 tumor cells from rats administered anti-DMPO in vivo. OKN-007 was also found to decrease nuclear factor erythroid 2-related factor 2, inducible nitric oxide synthase, 3-nitrotyrosine, and malondialdehyde in ex vivo F98 glioma tissues via immunohistochemistry, as well as decrease 3-nitrotyrosine and malondialdehyde adducts in vitro in F98 cells via ELISA. The results indicate that OKN-007 effectively decreases free radicals associated with glioma tumor growth. Furthermore, this method can potentially be applied toward other types of cancers for the in vivo detection of macromolecular free radicals and the assessment of antioxidants.

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Endogenous avidin biotin activity (EABA) in thyroid pathology: immunohistochemical study

Abstract: Among thyroid lesions, false positive reactions are highly probable in papillary thyroid carcinoma and in lymphocytic thyroiditis if immunohistochemical detection is used on systems containing (strept)avidin. The most probable reason is the high endogenous biotin content.

Cited by 11 publications

References 18 publications

Diagnostic value of neutrophil gelatinase-associated lipocalin (NGAL) immunoexpression in follicular-patterned lesions of the thyroid gland

Diagnostic value of neutrophil gelatinase-associated lipocalin (NGAL) immunoexpression in follicular-patterned lesions of the thyroid gland

When Tissue Antigens and Antibodies Get Along

OKN-007 decreases free radical levels in a preclinical F98 rat glioma model

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