Pericardium
Mary N. Sheppard in Practical Cardiovascular Pathology, 2022
Cancer may affect the pericardium directly (primary are rare or through metastases which are far more common). Cancer treatment (chemotherapy and radiotherapy) may affect the pericardium leading to pericarditis and myopericarditis. Pericardial effusions, tamponade and constrictive pericarditis are complications that can also occur. Any cancer can metastasize to the pericardium resulting in an effusion with the commonest cancers doing so being breast, lung and Hodgkin lymphoma. Primary pericardial tumours are rare, accounting for around 10% of all primary cardiac tumours with their prevalence in the general population being 0.001–0.007%. Secondary tumours or direct invasion into the pericardium is around 1000 times more common. Most are adenocarcinomas and form bulky nodular masses or encase the heart in a necrotic haemorrhagic pericardial thickening (Figs. 12.19a,b). The tumour infiltrates directly or by lymphatic permeation (Figs. 12.20a). Immunostaining is useful for diagnosis (Fig. 12.20b).
Amyloid Tumors of Breast, Lung and RIB in Nonsecretory Multiple Myeloma
Gilles Grateau, Robert A. Kyle, Martha Skinner in Amyloid and Amyloidosis, 2004
All specimens were fixed in PBS buffered formalin and embedded in paraffin. Deparaffinized sections (2-4 µm; 80°C for 14 to 16 hours) were stained with hematoxylin and eosin. The presence of amyloid was demonstrated by the appearance of green polarization color due to Congo red staining according to Romhanyi [6] in polarized light. Performic acid pretreatment [7] as well as potassium permanganate oxidation [8] with and without trypsin digestion served to demonstrate the resistance of the amyloid tumors. Glycosaminoglycans of the amyloid deposits were determined with the critical electrolyte concentration (CEC) method using the topooptical staining reaction with toluidine blue at pH 5,2 [5]. Immunostaining was performed with monoclonal antibodies directed against VS38 (1:50), IgA (1:10), IgG (1:100) and IgM (1:100) as well as the antibodies against the light chains kappa (1:500) and lambda (1:3000, all the preceding from Dako, Hamburg). Two different antisera against amyloid of lambda light chain (Aλ [HAR]; dilution 1:10000) and kappa light chain origin (Aκ [SIN]; dilution 1:50000) were used. Immunoreaction was visualized using the avidin-biotin complex.
Molecular testing of thyroid nodules
Demetrius Pertsemlidis, William B. Inabnet III, Michel Gagner in Endocrine Surgery, 2017
Using panels with more than one marker may increase the accuracy of ICC. In a small series of 115 indeterminate FNAB specimens evaluated by HBME-1 and CK-19, there were no false-negative ICC results. However, four false-positive cases occurred and specificity was 85%. Moreover, inconclusive ICC results were obtained in 38% of FNAB specimens, further limiting the described technique [19]. In another study of 50 indeterminate FNAB specimens evaluated by HBME-1, Gal-3, and p27 ICC, sensitivity was high (100%); however, specificity was again limited (86%). Colocalization of both Gal-3 and HBME-1 in the same cell was seen in 86% of the histologic malignancies and, when present, was 100% predictive of thyroid cancer [20]. Limitations of ICC include the higher cost incurred with the use of additional markers and variability in interpretation of immunostaining results. In addition, accurate results are reliant on obtaining enough cells by FNA alone, and some of the tests have been optimized on formalin-fixed, paraffin-embedded preparations that are not the current standard method for cytology smears used for routine diagnosis.
Gut Akkermansia muciniphila ameliorates metabolic dysfunction-associated fatty liver disease by regulating the metabolism of L-aspartate via gut-liver axis
Published in Gut Microbes, 2021
Yong Rao, Zhiqi Kuang, Chan Li, Shiyao Guo, Yaohao Xu, Dandan Zhao, Yutao Hu, Bingbing Song, Zhi Jiang, Zhenhuang Ge, Xiyuan Liu, Chengdao Li, Shuobin Chen, Jiming Ye, Zhishu Huang, Yongjun Lu
Immunoblotting and immunostaining were performed as previously described.6 Briefly, cells or tissues were lysed and total protein were extracted and quantified. Proteins were subjected to SDS-PAGE and transferred to PVDF membrane, and blocked with 5% bovine serum albumin (BSA) – Tris-buffered saline Tween for 30 min at room temperature, then membrane were incubated with primary antibodies overnight and followed secondary antibody incubation, protein bands were visualized with an ECL kit (Millipore). The uncropped data of immunoblotting were uploaded as a supplemented file. For Immunostaining, cells were fixed and blocked, then incubated with primary antibodies at 4°C overnight. After washing, cells were co-stained with fluorescent labeled secondary antibody and 2 μg/mL DAPI (for nucleus) at 37°C for 1 h. Fluorescence images were acquired using confocal microscope (Zeiss, Germany) with a 60× UPlanApoN oil immersion lens (NA 1.40).
Immunohistochemical evaluation of the prognostic and predictive power of epidermal growth factor receptor ligand levels in patients with metastatic colorectal cancer
Published in Growth Factors, 2020
Siavash Foroughi, Ryan A. Hutchinson, Hui-li Wong, Michael Christie, Ahida Batrouney, Rachel Wong, Margaret Lee, Jeanne Tie, Antony Wilks Burgess, Peter Gibbs
Staining intensity was scored independently by two observers (SF and RH) blinded to clinical information. The following IHC scoring scheme was used: membrane staining intensity was determined for tumour cells in a fixed field, where; 0, no staining; 1+, faint membranous reactivity; 2+, moderate membranous reactivity; and 3+, strong membranous reactivity. The percentage of cells at each staining level was recorded and a histological score (H-score) was then calculated using the following formula: (% of 1+ cells) × 1 + (% of 2+ cells) × 2 + (% of 3+ cells) × 3 (Pirker et al. 2012). All cases which were found to be discordant were rescored and discussed by both observers before a final score was given. Examples of immunostaining for each antibody are shown in Figure 1.
Thymosin β4 for the treatment of acute stroke: neurorestorative or neuroprotective?
Published in Expert Opinion on Biological Therapy, 2018
Daniel C Morris, Zheng G Zhang, Michael Chopp
Standard paraffin blocks were obtained from the center of the lesion, corresponding to coronal coordinates for bregma −1–1 mm. A series of 6 µm-thick sections at various levels (100 µm interval) were cut. Immunostaining was performed on these sections. Antibodies used for the identification of OPCs and OLs were NG-2 (chondroitin sulfate proteoglycan) (1:800, polyclonal rabbit, incubated overnight at 4ºC, Chemicon, CA) and APC (CC1) (adenomatous polyposis coli) (1:20, monoclonal mouse, incubated for 60 min at room temperature, Genway, CA), respectively. To identify proliferating OPCs and OLs, double immunostaining of NG2/BrdU and APC/BrdU was performed, respectively. Antibodies used for the identification of Aquaporin4 (AQ4) and endothelial cells were AQ4 (1:1500, polyclonal rabbit, incubated overnight at 4ºC, Millipore, Germany) and Endothelial Barrier antigen (EBA) (1:1000, monoclonal mouse, incubated overnight at 4ºC, Covance, NJ), respectively. Double immunolabeling was visualized by secondary antibodies conjugated to FITC and Cy3 (Jackson Immuno).