Staining Pattern
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2022 ◽  
Vol 17 (1) ◽  
Ye Zheng ◽  
Mingzhu Xu ◽  
Dong Zeng ◽  
Haitao Tong ◽  
Yuhan Shi ◽  

Abstract Aims Hepatitis B Virus (HBV) infection is the major risk factor for hepatocellular carcinoma (HCC) in East Asia. Here we aimed to further investigate the abundance of viral antigen and DNA within HBV-related HCC and surrounding tissues at histological level. Method In addition to routine histopathology, in situ hybridization (ISH) of HBV DNA and immunohistochemistry (IHC) of HBsAg were performed in tissues from 131 HBsAg-positive HCC patients undergoing liver resection. Serum α-fetoprotein together with basic biochemical and immunological parameter was also measured. Results Overall, the ISH of HBV DNA and IHC of HBsAg showed 31.3% and 92.9% positive rate respectively (p < 0.0001). The level of correlation between these two markers was much more significant in tumor (p < 0.0001) than in tumor-surrounding tissue (p = 0.01). HBsAg exhibited a much higher positive rate in tumor-adjacent tissue than in tumor tissue (86.6% versus 29.9%, p < 0.0001) with significantly different staining pattern. By contrast, the positive rate of HBV DNA ISH was comparable in tumor and surrounding tissue (17.6% versus 22.9%, p = 0.36). Yet the HBV DNA signal in tumor tissue showed predominant nuclear localization (87.0%) whereas staining pattern in adjacent tissue was mixed (43.3% nuclear localization, p = 0.0015). Finally, no significant association between intra-tumor HBV DNA/HBsAg positivity and major histological markers (microvascular invasion, tumor differentiation, etc.) or recurrence after surgery was observed. Conclusions These data confirmed the largely integrated state of HBV DNA, weaker expression and altered localization of surface antigen in tumor compared with surrounding tissue. The strikingly different prevalence and localization of HBsAg and HBV DNA reflected the complex and heterogeneous mechanisms leading to HBV-induced tumorigenesis.

2021 ◽  
Vol 10 (1) ◽  
pp. 4
Adepitan A. Owosho ◽  
Adeola M. Ladeji ◽  
Olufunlola M. Adesina ◽  
Kehinde E. Adebiyi ◽  
Mofoluwaso A. Olajide ◽  

Primary osteosarcomas of the jaw (OSJ) are rare, accounting for 6% of all osteosarcomas. This study aims to determine the value of SATB2 and MDM2 immunohistochemistry (IHC) in differentiating OSJ from other jawbone mimickers, such as benign fibro-osseous lesions (BFOLs) of the jaw or Ewing sarcoma of the jaw. Certain subsets of osteosarcoma harbor a supernumerary ring and/or giant marker chromosomes with amplification of the 12q13–15 region, including the murine double-minute type 2 (MDM2) and cyclin-dependent kinase 4 (CDK4) genes. Special AT-rich sequence-binding protein 2 (SATB2) is an immunophenotypic marker for osteoblastic differentiation. Cases of OSJ, BFOLs (ossifying fibroma and fibrous dysplasia) of the jaw, and Ewing sarcoma of the jaw were retrieved from the Departments of Oral Pathology and Oral Medicine, Faculty of Dentistry, Obafemi Awolowo University and Lagos State University College of Medicine, Nigeria. All OSJ retrieved showed histologic features of high-grade osteosarcoma. IHC for SATB2 (clone EP281) and MDM2 (clone IF2), as well as fluorescence in situ hybridization (FISH) for MDM2 amplification, were performed on all cases. SATB2 was expressed in a strong intensity and diffuse staining pattern in all cases (11 OSJ, including a small-cell variant, 7 ossifying fibromas, and 5 fibrous dysplasias) except in Ewing sarcoma, where it was negative in neoplastic cells. MDM2 was expressed in a weak to moderate intensity and scattered focal to limited diffuse staining pattern in 27% (3/11) of cases of OSJ and negative in all BFOLs and the Ewing sarcoma. MDM2 amplification was negative by FISH in interpretable cases. In conclusion, the three cases of high-grade OSJs that expressed MDM2 may have undergone transformation from a low-grade osteosarcoma (LGOS). SATB2 is not a dependable diagnostic marker to differentiate OSJ from BFOLs of the jaw; however, it could serve as a valuable diagnostic marker in differentiating the small-cell variant of OSJ from Ewing sarcoma of the jaw, while MDM2 may be a useful diagnostic marker in differentiating OSJ from BFOLs of the jaw, especially in the case of an LGOS or high-grade transformed osteosarcoma.

2021 ◽  
pp. 106689692110701
Hiroshi Minato ◽  
Akane Yoshikawa ◽  
Kazuyoshi Katayanagi ◽  
Hiroshi Kurumaya ◽  
Kaichiro Kato ◽  

Intrahepatic cholangiocarcinoma with rhabdoid morphology is rare, and only three case reports have been published to date, none of which discuss the genetic changes in the rhabdoid component. We present a case of intrahepatic cholangiocarcinoma with focal rhabdoid features and SMARCA4-deficiency detected using immunohistochemistry. A Japanese man in his 60s without viral hepatitis was diagnosed with an avascular tumor in the liver, measuring 4.4 cm in the greatest dimension. The tumor was mostly composed of moderately differentiated adenocarcinoma, focal poorly differentiated adenocarcinoma, and an undifferentiated rhabdoid component. Immunohistochemical analysis showed an inclusion-like staining pattern for keratin AE1/AE3 and vimentin in the rhabdoid component. BRG1/SMARCA4 was detected in the differentiated component but not in the poorly- and undifferentiated components. Our novel findings reflecting the morphological and genetic heterogeneity of intrahepatic cholangiocarcinoma and will aid the research on drugs targeting the aberrant SWItch/Sucrose NonFermentable complex.

2021 ◽  
Omondi Swaya Tyrus ◽  
Dedan Opondo ◽  
David O. Atandi ◽  
Benard Guyah ◽  
Ng’wena Gideon Magak

Abstract Background Prostate cancer is the leading cause of cancer-associated mortality in men. Most of the current biomarkers for detection of the disease have low sensitivity and specificity. Prostein is a newly reported prostate cancer biomarkers whose diagnostic utility can help in early detection of the disease. Nonetheless, previous studies have utilized limited number of samples to evaluate its immunohistochemistry (IHC) and reports on the African population are not available. The current study aimed to determine the prostein expression in archived prostatic core biopsies from prostate cancer patients in Western Kenya. Materials and Methods This was a retrospective study conducted on malignant and benign prostatic tissue core biopsies of 106 patients who underwent prostate core biopsy at Jaramogi Oginga Odinga Teaching and Referral Hospital and division of urology at Synergy Clinics, Kisumu between January 2018 to May 2021. Immunohistochemical technique was performed on each of the 106 samples and on the following non-prostatic male control biopsies; Testis, Penis, Liver and Esophagus. Cellular location of prostein staining was evaluated at X40, X100 and X400 magnification using a light microscope and was classified as cytoplasmic or nucleocytoplasmic. Intensity of prostein expression was assessed for each core biopsy at similar magnification and graded according the immunohistochemistry composite score. Results The biopsies had been obtained from men whose mean (SE) age was 72.00±0.93 years. 95.3% (101) of the biopsies were malignant and 4.7% (5) were benign. Four non-prostatic male tissues were included. 97% of malignant and all the benign prostate tissue stained positive for prostein whereas the four non-prostatic male tissues were negative. Staining intensities were weak (24.5%), Moderate (17.0%), strong (55.7%) and non-stained (2.8%). The staining was highly immunolocalized within the cytoplasm (95.1% cases) as compared to nucleocytoplasmic (2.0% cases). The mean immunoreactivity composite score was 1.91±0.96 (0.0-3.14). Strongly stained sections had a punctate plasma membrane staining pattern clustered within the cytoplasm in a perinuclear location whereas the weakly stained sections had faint and punctate coarse brown cytoplasmic granular appearing. Conclusion Prostein is exclusively expressed in benign and malignant prostate tissue with a higher cytoplasmic granular staining pattern in the present population. These findings suggest that prostein diagnostic utility is applicable in the current study population and routine IHC diagnosis of prostate cancer may be recommended.

2021 ◽  
pp. jnnp-2021-326656
Cristina Valencia-Sanchez ◽  
Andrew M Knight ◽  
M Bakri Hammami ◽  
Yong Guo ◽  
John R Mills ◽  

ObjectivesTo report the expanded neurological presentations and oncological associations of tripartite motif-containing protein 46 (TRIM46)-IgG seropositive patients.MethodsArchived sera/cerebrospinal fluid (CSF) were evaluated by tissue-based immunofluorescence assay to identify patients with identical axon initial segment (AIS)-specific staining pattern. Phage immunoprecipitation sequencing (PhIP-Seq) was used to identify the putative autoantigen.ResultsIgG in serum (17) and/or CSF (16) from 25 patients yielded unique AIS-specific staining on murine central nervous system (CNS) tissue. An autoantibody specific for TRIM46 was identified by PhIP-Seq, and autoantigen specificity was confirmed by transfected COS7 cell-based assay. Clinical information was available for 22 TRIM46-IgG seropositive patients. Fifteen were female (68%). Median age was 67 years (range 25–87). Fifteen (68%) patients presented with subacute cerebellar syndrome (six isolated; nine with CNS accompaniments: encephalopathy (three), brainstem signs (two), myelopathy (two), parkinsonism (one)). Other phenotypes included limbic encephalitis (three), encephalopathy with/without seizures (two), myelopathy (two). Eighteen (82%) had cancer: neuroendocrine carcinomas (9; pancreatic (3), small-cell lung (4), oesophagus (1), endometrium (1)), adenocarcinomas (6; lung (2), ovarian (2), endometrial (1), breast (1)), sarcoma (2) and gastrointestinal tumour (1). Neurological symptoms in three followed immune checkpoint inhibitor (ICI) administration.ConclusionsThis study supports TRIM46-IgG being a biomarker of paraneoplastic CNS disorders and expands the neurological phenotypes, oncological and ICI-related adverse event associations.

2021 ◽  
Conrad En-Zuo Chan ◽  
Ching-Ging Ng ◽  
Angeline P. C. Lim ◽  
Shirley Gek-Kheng Seah ◽  
De Hoe Chye ◽  

A human monoclonal antibody panel (PD4, PD5, PD7, SC23 and SC29) was isolated from the B cells of convalescent patients and used to examine the S protein in SARS-CoV-2-infected cells. While all five antibodies bound conformational-specific epitopes within SARS-CoV-2 Spike (S) protein, only PD5, PD7, and SC23 were able to bind to the Receptor Binding Domain (RBD). Immunofluorescence microscopy was used to examine the S protein RBD in cells infected with the Singapore isolates SARS-CoV-2/0334 and SARS-CoV-2/1302. The RBD-binders exhibited a distinct cytoplasmic staining pattern that was primarily localised within the Golgi complex and was distinct from the diffuse cytoplasmic staining pattern exhibited by the non-RBD binders (PD4 and SC29). These data indicated that the S protein adopted a conformation in the Golgi complex that enabled the RBD recognition by the RBD-binders. The RBD-binders also recognised the uncleaved S protein indicating that S protein cleavage was not required for RBD recognition. Electron microscopy indicated high levels of cell-associated virus particles, and multiple cycle virus infection using RBD-binder staining provided evidence for direct cell-to-cell transmission for both isolates. Although similar levels of RBD-binder staining was demonstrated for each isolate, the SARS-CoV-2/1302 exhibited slower rates of cell-to-cell transmission. These data suggest that a conformational change in the S protein occurs during its transit through the Golgi complex that enables RBD recognition by the RBD-binders, and suggests that these antibodies can be used to monitor S protein RBD formation during the early stages of infection.

2021 ◽  
Vol 11 (12) ◽  
pp. 1301
Arrigo Fruscalzo ◽  
Luigi Viola ◽  
Maria Orsaria ◽  
Stefania Marzinotto ◽  
Michela Bulfoni ◽  

Background: Recent reports indicate the potential role of the stimulated by retinoic acid 6 (STRA6) protein in developing insulin resistance. The study’s objective was to assess placental STRA6 expression and staining pattern in human pregnancy complicated by gestational diabetes mellitus (GDM). The expression pattern of further relevant genes involved in retinoid metabolism was also evaluated. Methods: A retrospective case–control study on paraffin-embedded placental tissue. Twenty-two human pregnancies affected by GDM, namely, 11 insulin-treated (iGDM) and 11 diet-controlled (dGDM), were compared with 22 normal-developed pregnancies (controls). An RT-PCR was performed in a random sample of 18 patients (six iGDM, six dGDM, and six controls) to assess RNA expression of STRA6 and further markers of retinoid metabolism. A semi-quantitative intensity evaluation at immunohistochemistry was performed for STRA6 in all 44 recruited patients. Results: STRA6 showed a decreased placental staining (9.09% vs. 68.18% positively stained samples, p < 0.05) and augmented RNA expression in dGDM patients than controls (ΔCT expression 0.473, IQR 0.403–0.566 vs. 0.149, IQR 0.092–0.276, p < 0.05). The protein staining pattern in patients affected by iGDM was comparable to controls. A reduced RNA expression of LPL, LRP1, VLDLR, and MTTP besides an augmented expression of LDLR was found in dGDM, while overexpression of LRP1 and LPL was found in iGDM patients. Unlike in the control group, significant positive correlations were found between RXRα and the proteins involved in the intracellular uptake of ROH, such as STRA6, LRP1, LRP2, and VLDLR. Conclusions: An altered placental expression and staining pattern of STRA6 were found in pregnancies complicated by GDM compared to the controls. These changes were coupled to an altered expression pattern of several other genes involved in the retinoid metabolism.

2021 ◽  
Vol 6 (4) ◽  
pp. 267-271
Faeeza Begum ◽  
Shaista Choudhary

Salivary gland neoplasm are rare and constitute about 3% of all head and neck neoplasms. Mucins are altered in pathological states and are stained by special stains like Periodic Acid Schiff, Alcian Blue and Mucicarmine. To study the histomorphology of resected salivary gland tumors and mucin staining pattern wherever indicated. Surgically resected specimens received at our tertiary care hospital and subjected to histopathological examination. Specimens were fixed in 10% formalin, processed and embedded in paraffin blocks, serially cut to get sections of 3-5 microns thickness. Stained with hematoxylin and eosin for all. Mucin stains were used wherever applicable. Total number of cases studied were 70. Out of which 46 were benign (65.7%) and 24 were malignant (34.3%). Among benign tumours, Pleomorphic adenoma was the commonest tumour (48.57), followed by Warthin tumor (7.14%), Basal cell adenoma (4.28%), Myoepithelioma (1.43%), Oncocytoma (1.43%), Hemangioma (1.43%), Sailolipoma (1.43%). The Mucoepidermoid carcinoma was the most common malignant tumor (17.14%) followed by Adenoid cystic carcinoma (5.71%), Acinic cell carcinoma (4.28%), Polymorphous adenocarcinoma (1.43%), Epithelial myoepithelial carcinoma, Squamous cell carcinoma (1.43%), Salioblastoma (1.43%), Lymphoma (1.43%). Parotid was most common site for both benign and malignant tumor. Females are affected more commonly than males. Mucin staining pattern was noted. Salivary gland tumors have complex range of morphological spectrum. Histopathological examination is the golden standard for diagnosis and mucin stain would add as an adjunct to the diagnosis.

2021 ◽  
Vol 11 (1) ◽  
Ea Kristine Clarisse Tulin ◽  
Chiaki Nakazawa ◽  
Tomomi Nakamura ◽  
Shion Saito ◽  
Naoki Ohzono ◽  

AbstractLectins are proteins with the ability to recognize and bind to specific glycan structures. These molecules play important roles in many biological systems and are actively being studied because of their ability to detect glycan biomarkers for many diseases. Hemagglutinin (HA) proteins from Clostridium botulinum type C neurotoxin complex; HA1, HA2, and HA3 are lectins that aid in the internalization of the toxin complex by binding to glycoproteins on the cell surface. HA1 mutants have been previously reported, namely HA1 W176A/D271F and HA1 N278A/Q279A which are specific to galactose (Gal)/N-acetylgalactosamine (GalNAc) and N-acetylneuraminic acid (Neu5Ac) sugars, respectively. In this study, we utilized HA1 mutants and expressed them in complex with HA2 WT and HA3 WT to produce glycan detecting tools with high binding affinity. Particularly, two types were made: Gg and Rn. Gg is an Alexa 488 conjugated lectin complex specific to Gal and GalNAc, while Rn is an Alexa 594 conjugated lectin complex specific to Neu5Ac. The specificities of these lectins were identified using a glycan microarray followed by competitive sugar inhibition experiments on cells. In addition, we confirmed that Gg and Rn staining is clearly different depending on cell type, and the staining pattern of these lectins reflects the glycans present on the cell surface as shown in enzyme treatment experiments. The availability of Gg and Rn provide us with new promising tools to study Gal, GalNAc, and Neu5Ac terminal epitopes which can aid in understanding the functional role of glycans in physiological and pathological events.

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