Journal of Histochemistry & Cytochemistry最新文献_第6页

Gelatin Zymography Can Be Performed on Fixed Brain Tissue. 明胶Zymography可以在固定的脑组织上进行。

IF 1.9 4区生物学

Journal of Histochemistry & Cytochemistry Pub Date : 2023-09-01 Epub Date: 2023-08-20 DOI: 10.1369/00221554231194118

Zhenzhou Chen, Dennis Chuang, Shanyan Chen, Qiwei He, Brittany N Tomlison, Jiankun Cui, Zezong Gu

{"title":"Gelatin Zymography Can Be Performed on Fixed Brain Tissue.","authors":"Zhenzhou Chen, Dennis Chuang, Shanyan Chen, Qiwei He, Brittany N Tomlison, Jiankun Cui, Zezong Gu","doi":"10.1369/00221554231194118","DOIUrl":"10.1369/00221554231194118","url":null,"abstract":"Gelatin zymography is widely used to detect gelatinase activity, which is performed on unfixed tissue because it is assumed that fixation inactivates enzymes. However, using fixed tissues has several advantages over using fresh tissues for such prevention of tissue decay, thereby preserving the proteins as well as the morphology and structure of the specimens. In this study, we investigated the effects of the four commonly used fixatives (ethanol, acetone, zinc-based fixative (ZBF), and paraformaldehyde (PFA)) on the gelatinolytic activity in mouse brain tissue. Multiple protocols were employed to extract proteins from the fixed brain tissue. Western blotting and in-gel zymography (IGZ) were used to detect the gelatinase proteins and gelatinolytic activity of the extractions, respectively. In situ zymography (ISZ) revealed that ethanol, acetone, ZBF, and short-time PFA fixation did not inhibit gelatinolytic activity. Neither 1% Triton + 1 M NaCl nor 10% DMSO + 1 M NaCl was effective in extracting proteins from ethanol-, acetone-, ZBF-, or PFA-fixed brain tissues. However, 8 M urea + 4% CHAPS effectively extracted gelatinase proteins from ethanol- and acetone-fixed tissues while retaining the gelatinolytic activity. 2% SDS effectively extracted gelatinase proteins from ethanol-, acetone-, and ZBF-fixed tissues while retaining the gelatinolytic activity. Although 2% SDS + heating extracted gelatinase proteins from ethanol-, acetone-, ZBF-, and even long-term PFA-fixed tissues, the gelatinolytic activity was not retained. Our findings suggest that both ISZ and IGZ can be performed on fixed brain tissue, which is anticipated to be an improvement over the conventionally used gelatin zymography methods. (J Histochem Cytochem 71: 481-493, 2023).","PeriodicalId":16079,"journal":{"name":"Journal of Histochemistry & Cytochemistry","volume":null,"pages":null},"PeriodicalIF":1.9,"publicationDate":"2023-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10501363/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10607267","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Reflections on the Applications of Immunohistochemistry to Understanding the Maintenance of Electrochemical Gradients in the Inner Ear. 应用免疫组织化学方法研究内耳电化学梯度的维持。

IF 1.9 4区生物学

Journal of Histochemistry & Cytochemistry Pub Date : 2023-09-01 Epub Date: 2023-08-03 DOI: 10.1369/00221554231190438

Bradley A Schulte

引用次数: 0

An Intermittent Cytochemist. 间歇性细胞化学家

IF 1.9 4区生物学

Journal of Histochemistry & Cytochemistry Pub Date : 2023-09-01 Epub Date: 2023-08-23 DOI: 10.1369/00221554231195393

Thoru Pederson

引用次数: 0

Two Transient Receptor Potential Channels at Focal Adhesions. 局灶性粘连处的两个瞬时受体电位通道。

IF 1.9 4区生物学

Journal of Histochemistry & Cytochemistry Pub Date : 2023-09-01 Epub Date: 2023-08-18 DOI: 10.1369/00221554231194119

Ioli Mitsou, Cathrine Rein Carlson, Hinke A B Multhaupt, Cord Brakebusch, John R Couchman

{"title":"Two Transient Receptor Potential Channels at Focal Adhesions.","authors":"Ioli Mitsou, Cathrine Rein Carlson, Hinke A B Multhaupt, Cord Brakebusch, John R Couchman","doi":"10.1369/00221554231194119","DOIUrl":"10.1369/00221554231194119","url":null,"abstract":"Recently there have been reports that identify two transient receptor potential channels in cell-matrix junctions known as focal adhesions. These are the calcium channel TRP canonical 7 and the calcium-activated monovalent ion channel, TRP melastatin (TRPM) 4. Here, we report on the occurrence of TRPM4 in focal adhesions of fibroblasts. Of three commercial antibodies recognizing this channel, only one yielded focal adhesion staining, while the other two did not. The epitope recognized by the focal adhesion-localizing antibody was mapped to the extreme C-terminus of the TRPM4 protein. The other two antibodies bind to N-terminal regions of the TRPM4 proteins. Deletion of the TRPM4 gene by CRISPR/cas9 techniques confirmed that this channel is a bona fide focal adhesion component, while expression of full-length TRPM4 proteins suggested that processing may occur to yield a form that localizes to focal adhesions. Given the reports that this channel may influence migratory behavior of cells and is linked to cardiovascular disease, TRPM4 functions in adhesion should be explored in greater depth. (J Histochem Cytochem 71: 495-508, 2023).","PeriodicalId":16079,"journal":{"name":"Journal of Histochemistry & Cytochemistry","volume":null,"pages":null},"PeriodicalIF":1.9,"publicationDate":"2023-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10501361/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10258431","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

5hmC Immunohistochemistry: A Predictor of TERT Promoter Mutational Status in Follicular Thyroid Carcinoma? 5hmC免疫组织化学:滤泡性甲状腺癌TERT启动子突变状态的预测因子?

IF 3.2 4区生物学

Journal of Histochemistry & Cytochemistry Pub Date : 2023-08-01 DOI: 10.1369/00221554231190437

Martin Hysek, Samuel L Hellgren, Vincenzo Condello, Yiyi Xu, Catharina Larsson, Jan Zedenius, C Christofer Juhlin

{"title":"5hmC Immunohistochemistry: A Predictor of TERT Promoter Mutational Status in Follicular Thyroid Carcinoma?","authors":"Martin Hysek, Samuel L Hellgren, Vincenzo Condello, Yiyi Xu, Catharina Larsson, Jan Zedenius, C Christofer Juhlin","doi":"10.1369/00221554231190437","DOIUrl":"https://doi.org/10.1369/00221554231190437","url":null,"abstract":"Telomerase reverse transcriptase (TERT) gene aberrancies correlate to adverse prognosis in follicular thyroid carcinoma (FTC). As loss of 5-hydroxymethylcytosine (5hmC) has been associated with TERT promoter mutations in papillary thyroid carcinoma, this study sought to analyze the levels of 5hmC in a cohort of follicular thyroid tumors with available TERT data. A total of 29 tumors (26 FTCs, 2 follicular thyroid tumors of uncertain malignant potential, and 1 oncocytic thyroid carcinoma) with known TERT promoter mutational status and TERT gene expression were assessed for 5hmC immunoreactivity using two antibodies (clones RM236 and 4D9.) Slides were analyzed using a semiquantitative scoring system. Of the 10 tumor cases with aberrant TERT, only 1 scored negative with both antibodies (1/10; 10%), whereas the remaining 9 cases (9/10; 90%) exhibited some positivity for at least one antibody. Of the 19 TERT wild-type tumors, no case was scored negative using RM236, and 2 cases (2/19; 11%) using 4D9. The differences between TERT promoter mutated and wild-type groups were non-significant. The sensitivity and specificity for 5hmC immunohistochemistry (IHC) to detect mutated cases were 10% and 100% (RM236) and 20% and 89% (4D9). Therefore, 5hmC IHC is not a sensitive marker for detecting TERT promoter mutations in follicular thyroid tumors.","PeriodicalId":16079,"journal":{"name":"Journal of Histochemistry & Cytochemistry","volume":null,"pages":null},"PeriodicalIF":3.2,"publicationDate":"2023-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10424576/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10005160","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

SSEA-1 Correlates With the Invasive Phenotype in Breast Cancer. SSEA-1 与乳腺癌侵袭性表型相关

IF 1.9 4区生物学

Journal of Histochemistry & Cytochemistry Pub Date : 2023-08-01 Epub Date: 2023-07-21 DOI: 10.1369/00221554231189312

Katharina T Kohler, Anna A Møller Hansen, Jiyoung Kim, René Villadsen

{"title":"SSEA-1 Correlates With the Invasive Phenotype in Breast Cancer.","authors":"Katharina T Kohler, Anna A Møller Hansen, Jiyoung Kim, René Villadsen","doi":"10.1369/00221554231189312","DOIUrl":"10.1369/00221554231189312","url":null,"abstract":"The glycan moiety Lewis X (LeX) has been implicated in defining progenitor cells as well as playing a role in the progression of solid tumors, including breast cancer. Here, we used the original stage-specific embryonic antigen-1 (SSEA-1) antibody, MC-480, targeting the LeX motif to examine the expression pattern of this marker within the context of a differentiation hierarchy as well as functional properties of breast cancer cells. Immunohistochemical staining revealed the presence of SSEA-1 in a progenitor zone in the normal breast gland. In breast cancer, 81 of 220 carcinomas (37%) were positive for SSEA-1 and a distinct pattern could be correlated to major subtypes. Specifically, estrogen receptor alpha (ERα)-negative tumors showed a higher frequency of SSEA-1 expression compared to ERα-positive tumors, which are generally considered more differentiated (56% vs 29%, p<0.005). Functional assays performed on two representative breast cancer cell lines demonstrated that SSEA-1-expressing cells exhibited cancer stem cell properties as well as having more invasive potential, regardless of ERα status. A potential role of SSEA-1 in metastasis was confirmed by pairwise staining of primary- and corresponding lymph node tumors. Altogether, our data suggest that expression of SSEA-1 in breast cancer contributes to the malignant phenotype.","PeriodicalId":16079,"journal":{"name":"Journal of Histochemistry & Cytochemistry","volume":null,"pages":null},"PeriodicalIF":1.9,"publicationDate":"2023-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10424578/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10004457","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Commentary on a Classic JHC Article on Intracellular Hyaluronan Associated With the Mitotic Spindle. JHC 经典文章《细胞内透明质酸与有丝分裂纺锤体的关系》评论。

IF 1.9 4区生物学

Journal of Histochemistry & Cytochemistry Pub Date : 2023-08-01 Epub Date: 2023-07-16 DOI: 10.1369/00221554231189309

Stephen P Evanko, Thomas N Wight

引用次数: 0

Altered Expression of Heme Oxygenase 2 in Heme Oxygenase 1-deficient Mouse Embryos. 血红素氧化酶1缺陷小鼠胚胎中血红素氧化酶2表达的改变。

IF 1.9 4区生物学

Journal of Histochemistry & Cytochemistry Pub Date : 2023-08-01 Epub Date: 2023-07-22 DOI: 10.1369/00221554231189310

Meenakshi Rana, Divya Bajaj, Pooja Choubey, Sidhant Jain, Sharmila Basu-Modak

{"title":"Altered Expression of Heme Oxygenase 2 in Heme Oxygenase 1-deficient Mouse Embryos.","authors":"Meenakshi Rana, Divya Bajaj, Pooja Choubey, Sidhant Jain, Sharmila Basu-Modak","doi":"10.1369/00221554231189310","DOIUrl":"10.1369/00221554231189310","url":null,"abstract":"Heme oxygenases (Hmoxs) are enzymes that catalyze the first and rate-limiting step in the degradation of heme to carbon monoxide, iron, and biliverdin. The two main isozymes, namely Hmox1 and Hmox2, are encoded by two different genes. Mutation of the Hmox1 gene in mice is known to cause extensive prenatal lethality, and limited information is available about the expression of Hmox proteins in developing mouse embryos. In this study, immunohistochemistry was used to perform a detailed investigation comparing Hmox proteins in Hmox1 wild-type and knockout (KO) mouse embryos collected from wild-type and heterozygous timed-matings. Western analysis for Hmoxs was also done in the organs of late-gestation embryos. The results demonstrated cytoplasmic and nuclear localization of Hmoxs in all the organs examined in wild-type embryos. Interestingly, Hmox2 immunoreactive protein signals were significantly low in most of the organs of mid- and late-gestation Hmox1-KO embryos. Furthermore, relative levels of Hmox2 were revealed to be significantly lower in the lung and kidney of late-gestation Hmox1-KO embryos by western analysis, which complemented the immunohistochemistry findings in these two organs. The current study provides detailed immunoexpression patterns of Hmox proteins in wild-type and Hmox1-KO mouse embryos in mid- and late-gestation.","PeriodicalId":16079,"journal":{"name":"Journal of Histochemistry & Cytochemistry","volume":null,"pages":null},"PeriodicalIF":1.9,"publicationDate":"2023-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10424577/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10004464","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

The Battle Against Quenching and Fading for Fluorescence: A Model of How to Evaluate the Fluorophore's Behavior. 对抗荧光的淬灭和褪色：如何评估荧光团行为的模型。

IF 1.9 4区生物学

Journal of Histochemistry & Cytochemistry Pub Date : 2023-07-01 Epub Date: 2023-07-07 DOI: 10.1369/00221554231185184

Gloria E Hoffman

引用次数: 0

SS18-SSX Antibody: A Useful Tool to Save Time and Reduce Costs in Synovial Sarcoma Diagnosis. Proposal of a Novel Diagnostic Algorithm. SS18-SSX 抗体：滑膜肉瘤诊断中节省时间和降低成本的有用工具。提出一种新的诊断算法。

IF 1.9 4区生物学

Journal of Histochemistry & Cytochemistry Pub Date : 2023-07-01 Epub Date: 2023-06-26 DOI: 10.1369/00221554231184287

Giulia Orlando, Federica Santoro, Alessandra Linari, Cristian Tampieri, Ludovica Verdun di Cantogno, Simone De Meo, Nicola Ratto, Giovanni Grignani, Mauro Papotti, Rebecca Senetta

{"title":"SS18-SSX Antibody: A Useful Tool to Save Time and Reduce Costs in Synovial Sarcoma Diagnosis. Proposal of a Novel Diagnostic Algorithm.","authors":"Giulia Orlando, Federica Santoro, Alessandra Linari, Cristian Tampieri, Ludovica Verdun di Cantogno, Simone De Meo, Nicola Ratto, Giovanni Grignani, Mauro Papotti, Rebecca Senetta","doi":"10.1369/00221554231184287","DOIUrl":"10.1369/00221554231184287","url":null,"abstract":"Synovial sarcoma is a rare malignant mesenchymal neoplasm mostly affecting young adults, characterized by a specific translocation which results in the fusion of the SS18 gene on chromosome 18 with one of the three highly homologous SSX genes on chromosome X. Its morphological diagnosis, especially in monophasic or poorly differentiated variants, can be challenging because histological features often overlap with other malignant mesenchymal tumors. Until recently, the differential diagnosis mostly relied on the use of cytogenetic or molecular analyses to detect the specific t(X;18)(p11;q11) translocation, thus virtually restricting its correct identification to referral centers with a high histological and molecular pathology workflow. The recently commercialized highly sensitive and fusion-specific SS18-SSX antibody has significantly improved the approach to these tumors, representing a relatively cheap and easy to access tool for synovial sarcoma diagnosis. Through a retrospective analysis of 79 synovial sarcomas and histological mimickers, this study confirms the usefulness of the SS18-SSX antibody in the diagnosis of synovial sarcoma, particularly focusing on its application in the pathological response evaluation after neoadjuvant treatment as well as its time- and cost-saving advantages. Finally, we here propose a new diagnostic algorithm to apply into the routine practice.","PeriodicalId":16079,"journal":{"name":"Journal of Histochemistry & Cytochemistry","volume":null,"pages":null},"PeriodicalIF":1.9,"publicationDate":"2023-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10363909/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9868504","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0