• Asian Pacific Journal of Cancer Prevention
• /
• v.15 no.16
• /
• pp.6983-6987
• /
• 2014

Background: Modified toluidine blue staining (MTBs) is a simple, inexpensive and time saving method to detect H. pylori in gastric biopsy specimens. As a metachromatic stain, it simultaneously highlights intestinal metaplasia, a gastric cancer precancerous lesion. The aim of this study was to assess the reliability of MTBs compared with hematoxylin-eosin (H&E) for H. pylori detection using immunoperoxidase staining as the gold standard. This technique would be beneficial for a routine diagnosis and confirmation of H. pylori eradication in developing countries where endoscopic-based approaches are dominant. Materials and Methods: Esophagogastroduodenoscopy with triple site gastric biopsies was undertaken in 207 dyspeptic patients at Thammasat University Hospital, Thailand between 1997 and 1999. H&E, MTBs and immunoperoxidase staining were applied to each specimen. The presence or absence of H. pylori with each stain was interpreted separately and the sensitivity, specificity, positive and negative predictive values of H&E and MTBs were calculated. Results: A total of 282 specimens from 207 patients were evaluated. Using immunoperoxidase staining, organisms were positive in 117 specimens (41%). MTBs proved almost equally sensitive as immunoperoxidase (99%) and significantly more sensitive than H&E (85%). It has comparable specificity (96% vs 96%), PPV (95% vs 94%), and NPV (99% vs 90%) to H&E, using immunoperoxidase staining as gold standard. MTBs compared with immunoperoxidase staining, is cheaper (2 USD vs 12 USD) and faster (20 min vs 16 hrs) compared to immunoperoxidase staining. Conclusions: MTBs is effective, economical and easy to use in daily practice for the detection of H. pylori in gastric biopsy specimens. In addition to saving time in evaluating H. pylori associated gastritis, with a high sensitivity and ability to demonstrate intestinal metaplasia, the technique may have a role in confirmation of H. pylori eradication for gastric cancer prevention in a developing country setting.

• Korean Journal of Clinical Laboratory Science
• /
• v.37 no.1
• /
• pp.47-55
• /
• 2005

These studies were done to know decalcification methods to reduce the time of decalcification for quick bone tissue diagnosis. When bone tissue was decalcified with 10 % formic acid at room temperature, decalcification and hematoxylin & eosin (H&E) stains were complete and satisfactory after 12 hours, but some of the tissue sections fell off during staining. In this way, decalcification, H&E stains were complete and satisfactory after 24 hours, 36 hours and 48 hours, tissue sections didn't fall off during staining. When bone tissue was decalcified with 10 % formic acid in a $60^{\circ}C$ paraffin oven, decalcification and H&E stains were complete and satisfactory after 6 hours, but some tissue sections fell off during staining. In this way, decalcification and tissue sections were complete, with no falling off during staining after 8 hours, 10 hours, 12 hours, 14 hours, 24 hours, or H&E stains were satisfactory from 8 hours to 12 hours, but H&E stains appeared to reddish nucleus after 14 hours and 24 hours. Bone tissue was decalcified with 10 % formic acid for 6 hours, 12 hours and 24 hours at DECAL machine frequencies of 15 Hz and 45 Hz, and for 6 hours, 12 hours and 24 hours at a DECAL machine frequency of 90 Hz. Decalcification and H&E stains were complete and satisfactory after 6 hours at the 15 Hz and 45 Hz DECAL settings. Some of the tissue sections fell off during staining at the 15 Hz DECAL machine setting. At the 90 Hz setting, decalcification, H&E stains, and tissue sections were complete and satisfactory with no falling off during staining after 4 hours. In this way, decalcification, H&E stains, and tissue section were complete and satisfactory with no falling off during staining after 12 hours, 24 hours at all machine settings. Bone tissue was decalcified with 10 % formic acid for 6 hours, 12 hours and 24 hours at $37^{\circ}C$ 3 hours, 6 hours and 12 hours at $45^{\circ}C$ and 1 hours, 5 hours and 10 hours at $60^{\circ}C$ with the RHS-1 machine setting at 60Hz. At the temperatures of $37^{\circ}C$, $45^{\circ}C$, and $60^{\circ}C$ decalcification, H&E stains, and tissue sections were complete and satisfactory, with no falling off during staining except for after 6 hours at $37^{\circ}C$. 3 hours, 1 hours, or decalcification, H&E stains, and tissue sections were complete and satisfactory with no falling off during staining after 12 hours and 24 hours at $37^{\circ}C$, 6 hours and 12 hours at $45^{\circ}C$, and 5 hours at $60^{\circ}C$. But H&E stains appeared to reddish nucleus after 10 hours at $60^{\circ}C$. From the above reults, the authors were able to deduce that decalcification is accelerated by heat and frequency. We therefore think that it is necessary for machines which are similar to the RHS-1 machine to be maintained at the temperature evenly with agitation effect for quick decalcification.

• Biomedical Science Letters
• /
• v.17 no.3
• /
• pp.197-202
• /
• 2011

Mitosis count is one of the most helpful morphologic features for distinguishing pulmonary typical carcinoid (TC) from atypical carcinoid (AC). However, identifying areas of highest mitotic activity is tedious and time-consuming, and mitosis count may vary substantially among pathologists. Anti-phosphohistone H3 (PHH3) is an antibody that specifically detects histone H3 only when phosphorylated at serine 10 or serine 28, an event that is concurrent with mitotic chromatin condensation and not observed during apoptosis. In this study, immunohistochemical staining for PHH3 was performed to determine whether PHH3 was a reliable and objective mitosis-specific marker for pulmonary carcinoid tumors. Seventeen cases of surgically resected pulmonary carcinoid tumors (12 TCs and 5 ACs) were obtained and classified according to the 2004 World Health Organization classification. Mitotic counts determined by PHH3 correlated to ones determined by hematoxylin and eosin (H&E) staining; however, PHH3 mitotic counts (mean mitotic counts: 1 in TCs and 3.2 in ACs) were slightly higher than H&E mitotic counts (mean mitotic counts: 0.25 in TCs and 1.8 in ACs). The mitotic counts determined by experienced observer were more correlated to those determined by inexperienced observer with the PHH3-based method (R=0.968, P<0.001) rather than H&E staining (R=0.658, P<0.001). These results suggest that the PHH3 mitotic counting method was more sensitive and simple for detecting mitoses compared to traditional H&E staining. Therefore, PHH3 immunohistochemistry may contribute to more accurate and reproducible diagnosis of pulmonary carcinoid tumors and may be a valuable aid for administrating appropriate clinical treatment.

• Journal of the Korea Academia-Industrial cooperation Society
• /
• v.20 no.8
• /
• pp.417-424
• /
• 2019

Despite its superior ability to show distinct cellular morphology and for long-term storage, conventional tissue fixation by formalin has many drawback, including slower fixation, the exposure to harmful chemicals and extensive protein modification. Herein, we assessed the effects of rapid microwave-assisted tissue fixation on histological examination and on protein integrity by comparing these microwave irradiation fixated tissues with the formalin-fixed tissues. One of the paired mouse tissues (liver and kidney) was fixed in formalin and the other was fixed by using microwave irradiation in phosphate buffered saline. Each slide from the paraffin-embedded tissues was examined by H & E staining for the adequacy of fixation and by immunohistochemical staining for antigenicity in a blinded fashion. Evaluation of protein recovery and the protein quality from the fixed tissues were analyzed by the BCA method and Western blotting, respectively. The results from H & E staining and immunohistochemical staining showed that the sections obtained from microwave-fixed tissues under our experimental conditions were comparable to those of the formalin-fixed tissues except for the integrity of RBCs. Furthermore, proteins were effectively extracted from the microwave-fixed tissues with acceptable preservation of the proteins' quality. Taken together, this microwave-assisted tissue processing yields a quick fixation and better protein recovery in higher amounts, as well as the adequacy of fixation and the antigenicity being comparable to formalin-fixed tissues, and this all suggests that this new fixation technique can be applied in an environment where rapid tissue fixation is required.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.33 no.3
• /
• pp.571-575
• /
• 2004

Escherichia coli O157:H7 cause hemorrhagic colitis and the extraintestinal complication of hemolytic-uremic syndrome, with their higher incidence occurring in children. Lipopolysaccharide (LPS) of E. coli O157:H7 is very important to make IgG anti-LPS with bactericidal activity. To identify the characteristic of E. coli OI57:H7, we isolated 60 MDa plasmid and amplified stx genes of shiga-like toxin (Stx) 1, 2 of E. coli O157:H7 by polymerase chain reaction (PCR) method. Using the simple purification method which contained phenol extract, ethanol precipitation and gel filtration steps, the LPS of E. coli O157:H7 was isolated and purified. Finally, we confirmed the purity of LPS through SDS-PAGE and silver nitrate staining.

• Journal of Life Science
• /
• v.22 no.8
• /
• pp.1034-1040
• /
• 2012

Synovium is the soft tissue that lines the non-cartilaginous surfaces within joints. It has been reported that synovial cells are activated during the pathogenesis of rheumatoid arthritis. In this study, we quantitate and compare the cellular composition of synovia derived from individuals with non-inflammatory osteoarthritis (OA) and those with inflammatory rheumatoid arthritis (RA). Synovia from OA (n=8) and RA (n=5) patients were used for hematoxylin and eosin (H&E) staining. A light microscopic examination has shown that RA synovia were morphologically thickened and hypertrophied as compared to OA synovia. We also performed an immunohistochemistry (IHC) analysis to classify cell types in the synovia using CD68, CD90, or PGP9.5 markers. As a result, we obtained quantitative data regarding the cell populations, which are macrophages in the lining layer and FLSs in the subintimal layer of the synovium. Further Photoshop analyses of the H&E images could allow the counting of the number and layer of the cells in the synovium. The number and layers of the macrophage cells were increased in the lining layer of the RA synovia as compared to the OA synovia. FLS cells also were increased in the subintimal layer of RA synovia. Therefore, quantification of the H&E stained images via Photoshop is a possible analysis protocol for synovium study. This quantitation also supports the idea that the increases in cell number and cell activation are important processes for RA pathogenesis.

• Journal of Applied Biological Chemistry
• /
• v.57 no.2
• /
• pp.113-122
• /
• 2014

A5E is complex of several medicinal herb ethanol extracts. The aim of this study is investigating the anticancer effect for non-small cell lung cancer. The antitumor effects of A5E on NCI-H460 were examined by regulation of cell proliferation, apoptosis, cell cycle arrest, mitochondrial membrane potential (${\Delta}{\Psi}_m$), and apoptosis-related protein. Cell proliferation was measured by MTS assay. Apoptosis induced by A5E was confirmed by Annexin V-fluorescein isothiocyanate (FITC)/Propidium Iodide (PI) staining, and cell cycle arrest was measured by PI staining. NF-${\kappa}B$ translocation was detected by immunofluorescence and MMP (${\Delta}{\Psi}_m$) was measured by JC-1 staining. The expression of extrinsic pathway molecules such as FasL and FADD were elevated, and procaspase-8 was processed by A5E. In addition, intrinsic pathway related molecules were altered. The Bcl-2 and Bcl-xl levels decreased, Bax increased, and cytochrome C was released. In addition, the mitochondrial membrane potential collapsed, and caspase-3 and poly-(ADP-ribose) polymerase were processed by A5E. Moreover, A5E affected the cellular survival pathway involving phosphatidylinositol 3-kinase (PI3K)/Akt and NF-${\kappa}B$. PI3K and Akt were downregulated, also NF-${\kappa}B$ expression was decreased, and nuclear translocalization was inhibited by A5E. These results suggested that A5E delays proliferation, inhibit cell cycle progression and induce apoptosis in human lung cancer cell. We conclude that A5E is a potential anticancer agent for human lung carcinoma.

• Journal of the Institute of Convergence Signal Processing
• /
• v.24 no.1
• /
• pp.76-81
• /
• 2023

To distinguish cancer cells from normal cells, H&E (Hematoxylin & Eosin) staining is required. Pathological staining requires a lot of money and time. Recently, a digital dyeing method has been introduced to reduce such cost and time. In this paper, we propose a novel digital pathology algorithms. The first algorithm is the Pair method. This method learns the dyed phase image and unstained amplitude image taken by FPM (Fourier Ptychographic Microscopy) and converts it into a dyed amplitude image. The second algorithm is the unpair method. This method use the stained and unstained fluorescence microscopic images for modeling. In this study, digital staining was performed using a generative adversarial network (GAN). From the experimental results, we noticed that both the pair and unpair algorithms shows the excellent performance.

• Clinics in Shoulder and Elbow
• /
• v.21 no.2
• /
• pp.87-94
• /
• 2018

Background: Popeye deformity is common after rupture of the biceps muscle's long head tendon. Herein, we report on histological changes in biceps brachii muscles following tenotomy of the long head biceps tendon. Methods: Twelve Sprague-Dawley rats (12-week-old) underwent tenotomy of the long head biceps tendon in the right shoulder. At postoperative weeks 4, 7, and 10, the operative shoulders were removed by detaching the biceps brachii muscle from the glenoid scapula and humerus; the opposite shoulders were removed as controls. H&E staining was performed to elucidate histological changes in myocytes. Oil-red O staining was performed to determine fatty infiltration. Myostatin antibody immunohistochemistry staining was performed as myostatin is expressed by skeletal muscle cells during myogenesis. Results: H&E staining results revealed no changes in muscle cell nuclei. There were no adipocytes detected. Compared with that of the control biceps, the cross-sectional area of the long head biceps was significantly smaller (p=0.00). Statistical changes in the total extent of the 100 muscle cells were significant (p=0.00). Oil-red O staining revealed no fatty infiltration. Myostatin antibody immunohistochemical staining revealed no significant difference between the two sides. Conclusions: Muscular changes after tenotomy of the long head biceps included a decrease in the size of the individual muscle cells and in relative muscle mass. There were no changes observed in muscle cell nuclei and no fatty infiltration. Moreover, there were no changes detected by myostatin antibody immunohistochemistry assay.

• Biomedical Science Letters
• /
• v.29 no.1
• /
• pp.48-52
• /
• 2023

Formaldehyde use is associated with serious health risks, which can affect medical personnel and technicians. Therefore, we investigated the efficacy of an alternative fixative, with respect to two types of formalin fixatives, by hematoxylin and eosin (H&E) staining, periodic acid Schiff (PAS) staining, immunohistochemical (IHC) staining, and RNA extraction. For H&E staining, the circular nucleus was stained dark blue by the basic dye hematoxylin and the cytoplasm was stained red by the acid dye eosin in all three fixative samples. No difference was found in the Duksan General Science (DGS), Sigma-Aldrich, and Core-Fix fixative samples (Corebiotech) used to fix kidney tissue, after PAS staining. IHC staining showed that CD4 was significantly increased in the lippolysaccharide (LPS)-treated group compared to the control group (vehicle), confirming the changes in specific molecules. The quantity and quality of RNA from tissues fixed in the three types of fixatives were evaluated. The average concentration of RNA was 106 ng/µL and average purity at A 260/280 ratio was 1.7~2.0, regardless of fixative used. For quality of protein, glyceraldehyde 3-phosphate dehydrogenase (GAPDH) protein was confirmed by Western blotting. In conclusion, Core-Fix can be used as a fixative for pathological tissues, in histological and molecular diagnoses.