scholarly journals Diagnosis of Primary Cutaneous Amyloidosis by Rapid 4,6-Diamidino-2-Phenylindole Staining

Dermatology ◽  
2021 ◽  
pp. 1-9
Author(s):  
Junchen Chen ◽  
Huan Yang ◽  
Zhijun Xu ◽  
Ping Lu ◽  
Liyan Yuan ◽  
...  
Keyword(s):  
Congo Red ◽  
Positive Staining ◽  
Dapi Staining ◽  
Prurigo Nodularis ◽  
Tissue Sections ◽  
Double Stranded Dna ◽  
Experience Levels ◽  
Standard Procedures ◽  
Staining Methods ◽  
Congo Red Staining

<b><i>Background:</i></b> Quick and accurate diagnosis of primary cutaneous amyloidosis (PCA) may be difficult because its symptoms are often subtle and nonspecific. <b><i>Objective:</i></b> We sought to review the literature on the roles of various staining methods in the diagnosis of amyloidosis and demonstrate added benefits of using rapid 4,6-diamidino-2-phenylindole (DAPI) staining in the diagnosis of PCA. <b><i>Methods:</i></b> Three groups of cases, namely, PCA, neurodermatitis, and prurigo nodularis, were retrieved from a computerized pathology database for study, and their paraffin-embedded tissue blocks were cut following standard procedures. The tissue sections were stained with three stains: hematoxylin-eosin (HE), Congo red, and DAPI stains, and examined under the microscope to compare the staining patterns of these three methods. We also performed amyloid keratin and apolipoprotein E (APOE) staining on the sections of PCA in order to further support our conclusion. The PCA sections were read by junior and senior dermatopathologists for comparison. <b><i>Results:</i></b> The sensitivity of DAPI staining for PCA was significantly higher than that of Congo red staining and HE staining (<i>p</i> &#x3c; 0.001). This statement holds true whether the experiment was grouped in one sample or was divided into groups of junior and senior dermatopathologists (<i>p</i> &#x3c; 0.001). The DAPI-positive staining areas, except for the nuclei, were consistent with the amyloid deposition areas. In this study, DAPI staining had a sensitivity of 98.6% and a specificity of 100%. <b><i>Conclusion:</i></b> DAPI staining could serve as a useful technique to establish the diagnosis of PCA, and its high efficacy in diagnosing PCA makes it less dependent on the experience levels of the evaluators. Additionally, the binding of DAPI to the A-T-rich sequence of double-stranded DNA suggests that amyloid may contain DNA or a similarly structured nucleic acid.

Feline Insular Amyloid: Histochemical Distinction from Secondary Systemic Amyloid

1981 ◽  
Vol 18 (2) ◽  
pp. 181-187 ◽  
Author(s):  
B. L. Yano ◽  
K. H. Johnson ◽  
D. W. Hayden
Keyword(s):  
Sulfuric Acid ◽  
Potassium Permanganate ◽  
Congo Red ◽  
Staining Procedure ◽  
Domestic Cats ◽  
Tissue Sections ◽  
Dilute Sulfuric Acid ◽  
Congo Red Staining ◽  
Red Dye ◽  
Human Primate

Amyloid in islets of Langerhans from 48 domestic cats, one human, one non-human primate, and one raccoon was compared with secondary systemic amyloid from three domestic cats, one dog, one human, and one cow to determine affinity for Congo red dye after treatment of paraffin-embedded tissue sections with potassium permanganate and dilute sulfuric acid. Insular amyloid from all six species was resistant to pretreatment with potassium permanganate, i.e., affinity for Congo red was retained, whereas secondary systemic amyloid from all species was sensitive to the potassium permanganate pretreatment. Other stains did not distinguish between insular and secondary systemic amyloid. The potassium permanganate-Congo red staining procedure thus can be used to differentiate insular from secondary systemic amyloid in the cat and other species. The results also indicate that insular amyloid and secondary systemic amyloid are of different chemical composition and pathogenesis.

SYSTEMIC AMYLOIDOSIS IN YOUNG MICE INDUCED BY HUMAN SALIVA ADMINISTRATION

2021 ◽  
pp. 40-56
Author(s):  
Vadim A. Kozlov ◽  
Vera Yu. Aleksandrova ◽  
Yulia V. Vasilyeva ◽  
Sergey P. Sapozhnikov ◽  
Pavel B. Karyshev
Keyword(s):  
Visual Field ◽  
Congo Red ◽  
Healthy Person ◽  
Human Saliva ◽  
Systemic Amyloidosis ◽  
Chronic Tonsillitis ◽  
Relative Area ◽  
Congo Red Staining ◽  
Number Of Cells ◽  
Young Mice

The urgency of the study lies in the fact that for the first time it was proved in the experiment that intraperitoneal administration of human saliva to young mice causes the development of systemic amyloidosis, comparable to hereditary systemic amyloidosis in humans, induced by mutagenic changes in saliva lysozyme. The aim of the research: to test human saliva as an amyloidogen in a model experiment on young mice in comparison with the albumin model of amyloidosis that we developed earlier. White male mice weighing 20.0–25.0 g were divided into six groups: Intact mice (5); 30 days 1 time every other day were intraperitoneally administered to groups: 1 (5) soy cream substitute TU 9199-004-58706213-10 15 10 g/100 ml water 0.1 ml/10 g; 2 (3) saliva of a healthy person (SHP) 0.5 ml; 3 (3) SHP and Ciprofloxacin (C) 0.05 mg/10 g of weight; 4 (3) saliva of a person with chronic tonsillitis (remission, SCT); 5 (3) SCT+C. Kidney weight changed from 176.0±21 mg in intact mice (IM) to 197±43,0, 195,0±18,0, 195,0±18,0, 183,0±44,0, 153,0±25,0 mg, respectively. The number of cells per visual field of the tubular epithelium decreased by 1.3, 1.2, 1.6, 1.4, 1.4 times from the number of cells per visual field in IM 380.0±84.0 (p = 0.00), glomerular cells – by 1.4, 1.1, 1.3, 1.2, 1.2 times from the number of glomerular cells in IM 34.0±11.0 (p = 0.02), in groups 1-5, respectively. The glomerular area decreased by 2.4, 2.7, 2.3, 2.4, 3.0 times from the average glomerular area in IM (3163.7±832.7 mm2, p = 0.0000). The relative area of amyloid lesion when stained with Congo red: 18.4±8.2% (group 1), 26,7±10,6, 35,0±11,9, 45,7±13,2, 63,6±14,0% (groups 1-5). The spleen mass and length increased from 133.0±16.0 mg (IM) by 1.7, 1.7, 1.8, 2.1, 1.7 times and from 17.0±2.0 mm (IM) by 1.2, 1.2, 1.3, 1.4, 1.2 times, the number of cells per visual field decreased from 1318±205 (IM) by 1.7, 2.0, 3.3, 2.1, 2.0 times, groups 1-5, respectively (p < 0.01). The relative area of amyloid lesion in Congo red staining: 11,4±9,4%, 28,2±16,0, 27,9±20,8, 20,9±12,2, 16,5±8,3% (groups 1-5). The liver mass changed from 1.4±0.15 g (IM) to 1,57±0,26, 1,45±0,18, 1,53±0,1, 1,71±0,3, 1,36±0,4 g, the number of cells per visual field decreased from 67.0±15.0 (IM) by 1.9, 1.4, 1.8, 2.5, 1.9 times, the area of hepatocyte nuclei – from 54.0±12.3 µm (IM) by 3.1, 3.6, 4.2, 3.1, 3.3 times (groups 1-5, respectively, p < 0.01). The relative area of amyloid lesion in Congo red staining: 6,8±4,6%, 15,3±11,0, 24,5±12,8, 9,8±8,5, 12,6±2,3% (groups 1-5). Conclusions: 1) human saliva when administered intraperitoneally causes severe systemic amyloidosis in young mice with damage to the liver, kidneys and the spleen, which manifests more by cytotoxic effect than the volume of amyloid deposition; 2) human saliva contains a more active amyloidogen than albumin in combination with fillers of the soy cream substitute formulation; 2) human saliva of a person with chronic tonsillitis in remission is more amyloidogenic than the saliva of a healthy person; 3) Ciprofloxacin, administered in a therapeutic dose during the amyloid model formation, moderately increases the severity of amyloid organ damage; 4) human saliva can be used to simulate amyloidosis in an experiment on young mice; 5) the liver may contain enzymatic systems that perform the function of amyloidoclasia.

scholarly journals Evaluation of Congo Red Staining Kit to Determine Proteinuria in Preeclampsia

2018 ◽  
Author(s):  
JL Rodríguez Chávez ◽  
EK Fuentes Gutiérrez ◽  
MJ AngelesVázquez ◽  
H Mendieta Zerón
Keyword(s):  
Congo Red ◽  
Congo Red Staining

scholarly journals Histochemical Differential Diagnosis and Polarization Optical Analysis of Amyloid and Amyloidosis

2006 ◽  
Vol 6 ◽  
pp. 154-168 ◽  
Author(s):  
M. Bély
Keyword(s):  
Differential Diagnosis ◽  
Congo Red ◽  
Early Recognition ◽  
Polarized Light ◽  
Optical Analysis ◽  
Early Start ◽  
Amyloid Deposits ◽  
Protein Fibrils ◽  
Congo Red Staining ◽  
Sensitive Method

Amyloidosis is characterized by extracellular deposition of protein fibrils of chemically heterogeneous composition. Early recognition and identification of amyloid deposits allows an early start of therapy, which may entail a better prognosis. Congo red staining according to Romhányi (1971) is a highly specific and sensitive method for early microscopic recognition of amyloidosis. The main and most important types of amyloidosis may be distinguished by classic histochemical methods of performate pretreatment according to Romhányi (1979), or by KMnO4oxidation according to Wright (1977) followed by Congo red staining and viewed under polarized light. Differences in the speed of breakdown (disintegration) of amyloid deposits according to Bély and Apáthy allow a more precise distinction of various types of amyloid.

A 44-Year-Old Male with Subacute Onset of Syncope

2016 ◽  
Author(s):  
Jeffrey A. Cohen ◽  
Justin J. Mowchun ◽  
Victoria H. Lawson ◽  
Nathaniel M. Robbins
Keyword(s):  
Autonomic Neuropathy ◽  
Autonomic Dysfunction ◽  
Congo Red ◽  
Sympathetic Function ◽  
Autonomic Testing ◽  
Congo Red Staining ◽  
Definition Of ◽  
Amyloid P ◽  
Chronic Course ◽  
Autoimmune Etiology

Syncope in a patient with orthostatic hypotension (OH) may indicate autonomic dysfunction. The definition of OH is presented. Clinical features of parasympathetic and sympathetic function are discussed. The differential of acute autonomic dysfunction includes a number of conditions. An autoimmune etiology may occur autoimmune autonomic ganglionopathy. Serologic testing can assist in this diagnosis. If autoimmune immune modulating therapies may be indicated. Autonomic neuropathy may be a paraneoplastuc syndrome. Autonomic testing can also help with documenting autonomic neuropathy as well as the whether the defects are predominately parasympathetic or sympathetic. Amyloid should be considered as should diabetes but both have a more chronic course. An appropriate tissue biopsy with Congo Red staining can help to confirm the diagnosis of amyloid.

scholarly journals Evidence of extrahepatic replication of hepatitis E virus in human placenta

2014 ◽  
Vol 95 (6) ◽  
pp. 1266-1271 ◽  
Author(s):  
Purabi Deka Bose ◽  
Bhudev Chandra Das ◽  
Rajib Kishore Hazam ◽  
Ashok Kumar ◽  
Subhash Medhi ◽  
...  
Keyword(s):  
Human Placenta ◽  
Hepatitis E Virus ◽  
Placental Tissue ◽  
Hepatitis E ◽  
Fetal Mortality ◽  
Positive Staining ◽  
Structural Protein ◽  
Tissue Sections ◽  
Extrahepatic Replication ◽  
Immunohistochemical Studies

The incidence and severity of hepatitis E virus (HEV) infection in pregnant women is high in developing countries. Transplacental transmission of HEV in the third trimester of pregnancy has been found to be associated with high fetal mortality. Based on this evidence and in the absence of reports on HEV replication in extrahepatic sites, this study was carried out to investigate if HEV replication occurs in the placenta of infected mothers. The study included 68 acute viral hepatitis (AVH) and 22 acute liver failure (ALF) pregnant patients. Viral RNA was extracted from blood and placenta. HEV replication in placenta was confirmed by a replicative negative-strand-specific reverse transcriptase PCR. Viral load was estimated by real-time PCR. Immunohistochemical studies were also carried out for in situ detection of HEV in placental tissue sections. Replicative HEV RNA was detectable only in the placenta in ALF and AVH cases and not in blood samples. Positive staining of placental tissue sections with HEV antibody against the viral structural protein ORF3 was observed. HEV replication in placenta also correlated with fetal and maternal mortality in ALF patients. It is demonstrated for the first time that HEV replication occurs in human placenta and that placenta may be a site of extrahepatic replication of HEV in humans.

scholarly journals Immunocytochemical localization of the posterior lobe hormones and of their carrier proteins.

1980 ◽  
Vol 28 (5) ◽  
pp. 469-471 ◽  
Author(s):  
F Vandesande ◽  
K Dierickx ◽  
N Goossens
Keyword(s):  
Staining Method ◽  
Posterior Lobe ◽  
Serial Sections ◽  
Double Staining ◽  
Carrier Proteins ◽  
Tissue Sections ◽  
Tissue Antigens ◽  
Staining Methods ◽  
Double Staining Method ◽  
The One

Serial sections of vertebrate hypothalami were stained with the immunocytochemical peroxidase-antiperoxidase method. In addition to the single staining method, our double staining method was used, which enabled us to visualize two tissue antigens in single tissue sections. In both staining methods, differentially adsorbed antineurohypophysial hormone sera, anti-somatostatin serum and anti-bovine neurophysin sera were used. The results confirm the one hormone, one neuron hypothesis.

scholarly journals Comparison of paired immunofluorescence and paired immunoenzyme staining methods based on primary antisera from the same species.

1982 ◽  
Vol 30 (6) ◽  
pp. 518-524 ◽  
Author(s):  
K Valnes ◽  
P Brandtzaeg
Keyword(s):  
Model Systems ◽  
Detection Sensitivity ◽  
Direct Immunofluorescence ◽  
Tissue Sections ◽  
Staining Methods ◽  
Pap Method ◽  
Blue Reaction ◽  
Antigen Antibody ◽  
Unlabeled Antibody ◽  
Sufficient Strength

Evaluation of sequential paired staining with the unlabeled antibody peroxidase-antiperoxidase (PAP) method showed that the brown reaction product of diaminobenzidine (DAB) concealed both enzyme and antigen-antibody sites in the reagent sequence. The blue reaction product of the alternative substrate, 4-chloro-1-naphthol (CN), exerted no such blocking effect. Hence, to avoid interactions between the two PAP sequences, DAB had to be used for the first and CN for the second antigen. Complete blocking required that the DAB color reaction be of sufficient strength. When two antigens were present in the same cell, the DAB deposits inhibited staining of the second antigen unless the brown color was decreased by progressive dilution of the initial primary antibody. A mixture of brown and blue could thus reflect either concomitant staining of the two antigens or unwanted interactions between the two PAP sequences. Double staining of individual cells was, therefore, equivocal and conclusions had to be based on comparative single staining results in adjacent tissue sections. Tests carried out in several model systems showed that paired direct immunofluorescence with fluorochrome conjugates of contrasting colors (green and red) was much less time-consuming, more reliable, and of higher detection sensitivity for analyses of unbalanced mixtures of two antigens in the same cell.

Immunohistochemistry of deparaffinised sections using antigen retrieval with microwave combined pressure cooking versus immunofluorescence in the assessment of human renal biopsies

2013 ◽  
Vol 66 (5) ◽  
pp. 374-380 ◽  
Author(s):  
Suozhu Shi ◽  
Ping Zhang ◽  
Qingli Cheng ◽  
Jie Wu ◽  
Jing Cui ◽  
...  
Keyword(s):  
Renal Biopsy ◽  
Tissue Structure ◽  
Antigen Retrieval ◽  
Renal Diseases ◽  
Positive Staining ◽  
Pressure Cooking ◽  
Tissue Sections ◽  
Simultaneous Evaluation ◽  
Long Term Preservation

BackgroundImmunofluorescence of frozen tissue sections (IF-F) is a traditional technique used in renal biopsy. However, IF-F has certain disadvantages, such as a few or even no glomeruli in the section, and limited long-term preservation of the fluorescently labelled samples.MethodsWe compared two-step immunohistochemistry (IHC) staining of deparaffinised sections for antigen retrieval with microwave combined high-pressure cooking to IF-F used to detect antigens of IgG, IgA, IgM, C3, C1q, κ and λ in patient renal biopsy samples. The number of glomeruli detected, sensitivity and specificity of positive staining, tissue structure, and location staining of the antigens were determined using the two methods in 285 patients diagnosed with different renal diseases.ResultsConcordant observations between IF-F and IHC were 99% for all antigen staining (1969 of 1995 observations) and 100% for IgG, IgA and IgM (all 285 observations). The number of glomeruli in IHC sections was significantly greater compared with IF-F sections (p<0.001). IHC provided clearer images of tissue structure, more precise localisation of positive-staining antigens, and IHC staining allowed simultaneous evaluation of tissue by light microscopy. Correlation between tissue structure and immune deposits are not readily attained by IF-F.ConclusionsIHC is superior to IF-F for immunopathological diagnosis of renal biopsy tissue and is a reliable replacement for the more traditional IF-F method.

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