scholarly journals COMPARATIVE HISTOCHEMICAL EVALUATION OF MUCINS EXPRESSION IN FETAL ESOPHAGUS AND ADENOCARCINOMAS OF THE GASTROESOPHAGEAL JUNCTION

2018 ◽  
Vol 40 (3) ◽  
pp. 223-227
Author(s):  
S Vernygorodskyi ◽  
T Rekun ◽  
P Zhuchenko

Aim: To study and compare the expression patterns of mucins in the fetal gastroesophageal junction (GEJ) region and adenocarcinomas of the GEJ in adults using histochemical method. Material and Methods: To reveal the expression of different mucins, tissue sections from formalin-fixed paraffin-embedded tissue blocks of fetal GEJ and carcinomas of GEJ were stained by the following histological stains: hematoxylin and eosin, periodic acid-Schiff (PAS), alcian blue (AB), pH 2.5, combined AB-PAS, and combined AB-Aldehyde Fuchsin (AB/AF). Results: Expression of mucins within the developing fetal GEJ has been demonstrated (gestational age 5–38 weeks). We studied histochemical features of neoplastic cells in carcinomas of GEJ from 90 adult patients. The results showed that in gastric type expression of neutral mucins dominated especially in well differentiated (G1) adenocarcinomas and corresponded to the last three months of GEJ development, while in the intestinal and mixed type significant acid mucins expression was detected in moderate (G2) and poorly (G3) differentiated adenocarcinomas and corresponded to 17–25 weeks of GA. No significant coincidence in mucins expression was observed in signet ring cell carcinomas in relation to GA. Conclusion: Our data suggest that decrease of neutral mucins level can be considered reliable phenotypic marker of poor prognosis of GEJ adenocarcinomas. Study of the developmental expression of mucin genes may improve understanding of the malignant transformation of esophageal tissue.

2016 ◽  
Vol 55 (1) ◽  
pp. 313-320 ◽  
Author(s):  
Taiichiro Kobayashi ◽  
Koji Watanabe ◽  
Hideaki Yano ◽  
Yukinori Murata ◽  
Toru Igari ◽  
...  

ABSTRACT Entamoeba histolytica is not a common causative agent of acute appendicitis. However, amoebic appendicitis can sometimes be severe and life threatening, mainly due to a lack of awareness. Also, its frequency, clinical features, and pathogenesis remain unclear. The study subjects were HIV-1-infected individuals who presented with acute appendicitis and later underwent appendectomy at our hospital between 1996 and 2014. Formalin-fixed paraffin-embedded preserved appendix specimens were reexamined by periodic acid-Schiff (PAS) staining and PCR to identify undiagnosed amoebic appendicitis. Appendectomies were performed in 57 patients with acute appendicitis. The seroprevalence of E. histolytica was 33% (14/43) from the available stored sera. Based on the medical records, only 3 cases were clinically diagnosed as amoebic appendicitis, including 2 diagnosed at the time of appendectomy and 1 case diagnosed by rereview of the appendix after the development of postoperative complications. Retrospective analyses using PAS staining and PCR identified 3 and 3 more cases, respectively. Thus, E. histolytica infection was confirmed in 9 cases (15.8%) in the present study. Apart from a significantly higher leukocyte count in E. histolytica -positive patients than in negative patients (median, 13,760 versus 10,385 cells/μl, respectively, P = 0.02), there were no other differences in the clinical features of the PCR-positive and -negative groups. In conclusion, E. histolytica infection was confirmed in 9 (15.8%) of the appendicitis cases. However, only 3, including one diagnosed after intestinal perforation, were diagnosed before the present analyses. These results strongly suggest there is frequently a failure to detect trophozoites in routine examination, resulting in an underestimation of the incidence of amoebic appendicitis.


2021 ◽  
Vol 42 (5) ◽  
pp. 2813-2824
Author(s):  
Monica Regina de Matos ◽  
◽  
Aline Patrícia Grzegozevski ◽  
Alessandra da Cruz ◽  
Arthur Colombari Cheng ◽  
...  

The genus Brachyspira corresponds to the group of bacteria formerly classified into the genus Serpulina and includes several commensal and pathogenic intestinal spirochetes that affect pigs, poultry, and other animal species, including humans. In birds, some pathogenic species of this genus causes a condition known as avian intestinal spirochetosis, which remains underdiagnosed, thereby causing serious economic losses. Brachyspira is a fastidious organism that necessitates the employment of fast and efficient identification techniques. The aim of this study was to identify Brachyspira spp. using histology, immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH) in formalin-fixed paraffin embedded (FFPE) tissue samples from the cecum of commercial poultry. Samples were collected from 129 birds aged between 35 and 45 days from commercial broiler farms. For evaluation, routine histology processing (H&E) and the histochemical technique, periodic acid–Schiff (PAS) were done. Additionally, FFPE tissue samples were evaluated for FISH and IHC. The histological lesions were analyzed and graded after H&E staining, and the goblet cells were counted and compared using PAS staining with the positive and negative samples obtained through FISH and IHC. For FISH, probes labeled with Brachyspira spp., B. pilosicoli, B. hyodysenteriae, and B. intermedia were used, whereas rabbit polyclonal antibody specific for Brachyspira spp. was used for IHC. Of 129 samples, 82 were positive with IHC and 86 were positive with FISH. The samples positive for the genus Brachyspira in the FISH technique were tested for B. pilosicoli, B. hyodysenteriae, and B. intermedia in which 56 were positive for B. pilosicoli, 75 for B. hyodysenteriae and 80 for B. intermedia. There was an increase in goblet cells in the samples positive for FISH and IHC. The techniques used were effective and gave corresponding results, thus serving as a fast and efficient tool for diagnosis.


2020 ◽  
Author(s):  
Zhiyong Liu ◽  
Zhonghua Sun ◽  
Hongyun Liu ◽  
Weipin Niu ◽  
Xin Wang ◽  
...  

Abstract Background: Adenomyosis (AM) is a common benign chronic gynaecological disorder; however, the precise pathogenesis of adenomyosis is still poorly understood. Single-cell RNA sequencing (scRNA-seq) can uncover rare subpopulations, explore genetic and functional heterogeneity, and reveal the uniqueness of each cell. It provides us a new approach to reveal biological issues from a more detailed and microscopic perspective. Here, we utilize this revolutionary technology to identify the changes of gene expression patterns between ectopic lesions and the eutopic endometrium at the single-cell level and explore a potential novel pathogenesis of AM.Methods: A control endometrium (sample with leiomyoma excluding endometrial disorders, n=1), eutopic endometrium and ectopic lesion (from a patient with adenomyosis, n=1) samples were analysed by scRNA-seq, and additional leiomyoma (n=3) and adenomyosis (n=3) samples were used to confirm colocalization and vasculogenic mimicry (VM) formation. Protein colocalization was visualized by immunofluorescence, and CD34-periodic acid-Schiff (PAS) double staining was used to assess the formation of VM.Results: The scRNA-seq results suggest that cancer-, cell motility- and inflammation- (CMI) associated terms, cell proliferation and angiogenesis play important roles in the progression of AM. Moreover, the colocalization of EPCAM and PECAM1 increased significantly in the ectopic endometrium group (P < 0.05), cell subpopulation with high copy number variation (CNV) levels possessing tumour-like features existed in the ectopic lesion sample, and VNN1- and EPCAM-positive cell subcluster displayed active cell motility in endometrial epithelial cells. Furthermore, the epithelial cells transformed to endothelial cells with the obvious accumulation of vasculogenic mimicry formations (positively stained with PAS but not CD34, P < 0.05) in ectopic lesions.Conclusions: In the present study, our results support the theory of adenomyosis derived from the invasion and migration of the endometrium. Moreover, cell subcluster with high CNV level and tumour-associated characteristics is identified. Furthermore, epithelial-endothelial transition (EET) and the formation of VM in tumours, the latter of which facilitates the blood supply and plays an important role in maintaining cell growth, were also confirmed to occur in AM. These results indicated that the inhibition of EET and VM formation may be a potential strategy for AM management.


1993 ◽  
Vol 41 (8) ◽  
pp. 1217-1221 ◽  
Author(s):  
J A Reed ◽  
B A Hemann ◽  
J L Alexander ◽  
D J Brigati

We report the rapid (less than 1 hr), immunocytochemical identification of various fungi in formalin-fixed, paraffin-embedded tissues using antisera originally developed for use in immunodiffusion assays. Primary antisera directed towards fungal genera including Aspergillus, Blastomyces, Candida, Coccidioides, Cryptococcus, Histoplasma, and Sporothrix were examined. The specificity of each antiserum was evaluated by the presence or absence of crossreactivity with other morphologically similar fungi in both paraffin-embedded pure fungal cultures and tissues with culture-confirmed fungal infections. Each antiserum reacted strongly with the fungus to which it had been raised, whether examined in pure culture or infected tissues. The antisera raised against Candida, Cryptococcus, and Sporothrix did not exhibit cross-reactivity with any other fungus tested. However, the antisera raised to Aspergillus, Blastomyces, Coccidioides, and Histoplasma demonstrated significant crossreactivity with other genera of fungi, thus precluding their routine use in diagnostic immunocytochemistry. The results indicate that immunocytochemistry may provide an important adjunct to other methods, such as immunodiffusion or complement fixation assays and histochemical stains such as the Grocott methenamine silver or periodic acid-Schiff, when attempts are made to specifically identify certain fungi in formalin-fixed, paraffin-embedded tissues before mycology culture results are available.


2009 ◽  
Vol 24 (4) ◽  
pp. 267-275 ◽  
Author(s):  
Ronaldo Nonose ◽  
Ana Paula Pimentel Spadari ◽  
Denise Gonçalves Priolli ◽  
Felipe Rodrigues Máximo ◽  
José Aires Pereira ◽  
...  

PURPOSE: To quantify the intensity of the expression of neutral and acids mucins in mucosa of the colon with and without fecal stream and to correlate this with the duration of fecal transit diversion. METHODS: Thirty male Wistar rats were subjected to fecal transit deviation in the left colon by a proximal colostomy and a distal mucous fistula. The animals were divided into three experimental groups, according to whether sacrificing would be performed six, 12 or 18 weeks after surgery. The expression of neutral and acid mucins was evaluated using the histochemical techniques of Periodic Acid Schiff and Alcian Blue, respectively. The tissue mucins expression was quantified by computer-assisted image analysis software (NIS-Elements) in the segments with and without fecal stream. Student's paired t test was used to compare the quantities of mucins in colon with or without fecal stream and variance between the experimental groups by ANOVA and Newman-Keuls post-test, establishing level of signification of 5% (p<0.05). RESULTS: There were significant decreased quantities of acid and neutral mucins in the colon without transit, compared with the colon with fecal stream, independent of the duration of exclusion. There was increased expression of neutral mucins in the colon with fecal stream after 12 and 18 weeks of exclusion. There was no increase in the expression of acid mucins in the colon with transit as the duration of fecal transit exclusion progressed. There was increased production of acid mucins in the animals submitted to diversion of the fecal stream for 18 weeks, compared with those subjected to diversion for 6 and 12 weeks. In the colon without fecal stream, there was increased expression of neutral mucins after 12 and 18 weeks of exclusion. CONCLUSIONS: Deviation of the fecal stream decreased the expression of acid and neutral mucins in the segments without fecal transit, compared with segments with transit. Regardless of the reduced expression of acid and neutral mucins in the segments without fecal stream, their tissue expression increased with increasing duration of intestinal deviation.


2019 ◽  
Vol 36 (E) ◽  
pp. 21-30
Author(s):  
Maria Luiza Assis Rodrigues ◽  
Sirlene Souza Rodrigues Sartori ◽  
Priscila Izabel Santos Totaro ◽  
Sérgio Luis Pinto da Matta

The ingestion of considerable amounts of water or food contaminated with nickel can be very toxic. The present work was conducted aiming to evaluate the effects of nickel exposures on ascending colon of adult Wistar male rats at hystometric level. We used 12 animals that were divided in a control (ingested uncontaminated water) and a nickel-contaminated (i.e., 25 mg de nickel/L of water) groups. Nickel chloride was offered in declorinated water and the experiment had a 56 days exposure period. A portion of the ascending colon was removed of the animals and subjected to hystological labelling processes using blue toluidin (for general hystometric description), Alcian Blue (AB, for acid mucins) and periodic acid-Schiff (PAS) technique (for neutral mucins). The potential differences between groups were desgined by applying the Whitney test and t test (p < 0.05). The crypts were smaller for the nickel-contaminated group, even though these organism exhibited broader and higher crypts. Nickel-contaminated animals exhibited a smaller amount of calyceform cells with AB and PAS positive reactions as well as a less mucus quantities when compared with nickel-uncontaminated animals. Such reductions on the amount of calyceform cells with AB and PAS positive reactions may be related wiht the shallower crypts, which possibly reduced the synthesis and secretion of mucins, compromissing the functional aspects (e.g., lubrification and intestinal mucosa protection) of the nickel-contaminated large intestines. Interestingly, the wider and higher crypts and higher epithelium collumn on the nickel-contaminated animals may represent a relevant trade-off for the intestinal mucosa protection.


2014 ◽  
Vol 6 (02) ◽  
pp. 076-079 ◽  
Author(s):  
Monalisa Hui ◽  
Megha S Uppin ◽  
Aruna K Prayaga ◽  
Sree Bhushan Raju ◽  
Liza Rajasekhar

ABSTRACT Background: Membranous nephropathy (MN) is the most common cause of nephropathy in adults. The diagnosis is based on characteristic light microscopic, electron microscope and immunofluorescence (IF) findings. In early MN, the light microscopic findings may be difficult to differentiate from minimal chain disease. In the absence of fresh frozen tissue for IF, immunohistochemistry with C4d aids in the diagnosis. Materials and Methods: A total 48 cases of MN diagnosed on renal biopsy were analyzed. The formalin fixed paraffin embedded tissues were stained with routine hematoxylin and eosin stains along with periodic acid-Schiff and silver methenamine stains to highlight the basement membrane. Fresh frozen tissues were available for IF in 40 cases. Immunostaining with C4d was done on paraffin-embedded sections by polymer-Horse Radish Peroxidase (HRP) technique using polyclonal antiserum to C4d (Biogenex, India). Results: There were 25 cases of idiopathic MN, 17 cases of Class V lupus nephritis and 2 cases were secondary to hepatitis C infection with cirrhosis. The glomerular basement membrane (GBM) was diffusely thickened with formation of spikes in 28 cases. In 11 cases the capillary loops were rigid but spikes were not seen and in 9 cases there was no apparent thickening of the basement membrane. All the cases showed diffuse positivity for C4d along the GBM. Conclusion: C4d is a reliable method to establish the diagnosis of MN and also a sensitive marker of complement activation reflecting the pathogenesis of MN.


1994 ◽  
Vol 31 (4) ◽  
pp. 455-461 ◽  
Author(s):  
A. J. Berrington ◽  
K. Jimbow ◽  
D. M. Haines

Seven monoclonal antibodies (Moabs), recognizing melanoma-associated antigens in human tissues, were evaluated for their ability to immunohistochemically stain formalin-fixed, paraffin-embedded canine melanomas. Only 2 Moabs, designated human melanosome-specific antigen (HMSA)-1 and HMSA-5, stained routinely processed canine melanomas, staining 21/35 (60%) and 24/35 (69%), respectively. Twenty-nine of 35 (83%) melanomas tested were stained if results of the 2 Moabs were combined. Monoclonal antibody HMSA-1 also stained neoplastic cells of 10/35 (29%) tumors of nonmelanocytic origin and some neurons and salivary gland epithelial cells in normal canine tissues. However, Moab HMSA-1 staining in the nonmelanocytic tumors, consisting of small, discrete periodic acid-Schiff-positivc cytoplasmic droplets, was readily distinguishable from the diffusely granular, cytoplasmic staining of melanocytic tumors. In addition to melanomas, Moab HMSA-5 stained melanocytes and some melanin-containing tumor cells of a pigmented basal cell tumor and melanocytes in normal canine skin. Monoclonal antibodies HMSA-1 and HMSA-5 immunohistochemically identified the majority of canine melanomas, had limited and distinguishable staining in normal tissues and nonmelanocytic tumors, and therefore may be a useful adjunct for the diagnosis of canine melanoma in formalin-fixed, paraffin-embedded tissues.


1962 ◽  
Vol 10 (1) ◽  
pp. 8-13 ◽  
Author(s):  
E. EDWARD EVANS ◽  
SIDNEY P. KENT

A procedure was described for demonstrating acidic polysaccharides in formalin-fixed paraffin-embedded tissue sections using fluorescein-labelled Aspergillus polysaccharide. This material is a basic polysaccharide composed of units of galactosamine and N-acetyl galactosamine and is synthesized by the mold Aspergillus parasiticus. The distribution of acidic polysaccharides shown by the fluorescein-labelled Aspergillus polysaccharide method was similar to results with the alcian blue-periodic acid Schiff sequence and to those with fluorescein labelled deacetylated chitin. Both epithelial and connective tissue mucins were demonstrated by the fluorescein-labelled basic polysaccharide. Aspergillus polysaccharide may be considered a colorless polyvalent cation that acts as a carrier for the fluorochrome in this procedure. Since it can react with many types of biological cells as well as with various substances isolated from cells, it may have other important applications in histochemistry and cytochemistry.


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