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Original Article
- Proposal of an Appropriate Decalcification Method of Bone Marrow Biopsy Specimens in the Era of Expanding Genetic Molecular Study
- Sung-Eun Choi, Soon Won Hong, Sun Och Yoon
- J Pathol Transl Med. 2015;49(3):236-242. Published online May 15, 2015
- DOI: https://doi.org/10.4132/jptm.2015.03.16
- 12,967 View
- 269 Download
- 41 Web of Science
- 42 Crossref
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Abstract
PDF - Background
The conventional method for decalcification of bone specimens uses hydrochloric acid (HCl) and is notorious for damaging cellular RNA, DNA, and proteins, thus complicating molecular and immunohistochemical analyses. A method that can effectively decalcify while preserving genetic material is necessary. Methods: Pairs of bilateral bone marrow biopsies sampled from 53 patients were decalcified according to protocols of two comparison groups: EDTA versus HCl and RDO GOLD (RDO) versus HCl. Pairs of right and left bone marrow biopsy samples harvested from 28 cases were allocated into the EDTA versus HCl comparison group, and 25 cases to the RDO versus HCl comparison group. The decalcification protocols were compared with regards to histomorphology, immunohistochemistry, and molecular analysis. For molecular analysis, we randomly selected 5 cases from the EDTA versus HCl and RDO versus HCl groups. Results: The decalcification time for appropriate histomorphologic analysis was the longest in the EDTA method and the shortest in the RDO method. EDTA was superior to RDO or HCl in DNA yield and integrity, assessed via DNA extraction, polymerase chain reaction, and silver in situ hybridization using DNA probes. The EDTA method maintained intact nuclear protein staining on immunohistochemistry, while the HCl method produced poor quality images. Staining after the RDO method had equivocal results. RNA in situ hybridization using kappa and lambda RNA probes measured RNA integrity; the EDTA and RDO method had the best quality, followed by HCl. Conclusions: The EDTA protocol would be the best in preserving genetic material. RDO may be an acceptable alternative when rapid decalcification is necessary. -
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Case Report
- Pulmonary Calciphylaxis Associated with Acute Respiratory and Renal Failure Due to Cryptogenic Hypercalcemia: An Autopsy Case Report
- Na Rae Kim, Jin Won Seo, Young Hwan Lim, Hyoung Suk Ham, Wooseong Huh, Joungho Han
- Korean J Pathol. 2012;46(6):601-605. Published online December 26, 2012
- DOI: https://doi.org/10.4132/KoreanJPathol.2012.46.6.601
- 7,497 View
- 45 Download
- 5 Crossref
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Abstract
PDF
Metastatic calcification is rare; it is found during autopsy in patients who underwent hemodialysis. Diffuse calcium precipitation of small and medium-sized cutaneous vessels, known as calciphylaxis, can result in progressive tissue necrosis secondary to vascular calcification. This condition most commonly involves the skin; however, a rare occurrence of visceral calciphylaxis has been reported. Here we report on an autopsy case. Despite a thorough evaluation, and even performing an autopsy, the underlying cause of acute-onset hypercalcemia, resulting in the production of pulmonary calciphylaxis and metastatic renal calcification associated with acute respiratory and renal failure, could not be determined. Metastatic calcification often lacks specific symptoms, and the degree of calcification is a marker of the severity and chronicity of the disease. This unusual autopsy case emphasizes the importance of rapidly progressing visceral calciphylaxis, as well as its early detection.
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Citations
Citations to this article as recorded by- Acute Respiratory Distress Syndrome Induced by Parathyroid Storm
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Original Article
- Calcifying Synovial Sarcoma.
- Nam Bok Cho, Tae Jin Lee, Mi Kyung Kim, Yong Wook Park, Kye Yong Song
- Korean J Pathol. 1995;29(4):536-539.
- 1,428 View
- 21 Download
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Abstract
PDF
- A case of calcifying synovial sarcoma, occuring in the deep muscle fascia of the left thigh is reported. The presence of extensive calcification in synovial sarcoma is a favorable sign for prognosis. The patient was a 31-year-old female. The tumor mass had been present for 3 years, accompanying local tenderness. X-ray revealed a soft tissue tumor with central calcification, which was located between the adductor magnus and brevis of the left thigh. The tumor size was 7 x 5.5 cm. There was no connection with the knee joint or the femur. Grossly, the tumor was a relatively well circumscribed hard tumor with massive calcification. Microscopically, the tumor was composed of predominantly spindle cells with accompanying hyalinization, numerous spherical concretions and ossification. The epithelial component was not clearly noted. Mitotic figures were rarely noted in the densely cellular area. Immunohistochemical staining for EMA, S-100, vimentin, and carcinoembryonic antigen was negative while vimentin and cytokeratin were weakly and focally positive. EM study revealed multiple desmosome-like calcification intercellular junctions with a slit-like lumen and an incomplete basal lamina, which suggest that tumor show these cells were undergoing epithelia] differentiation. Above evidence suggest that this tumor is a synovial sarcoma associated with a large area of calcification, a so called calcifying synovial sarcoma. No recurrence was noted in one and half years of follow up.
Case Report
- Fine Needle Aspiration Cytology of Salivary Duct Carcinoma with Calcification in Submandibular Gland: A Case Report .
- Ki Jung Yun, Weon Cheol Han, Hyang Jeong Jo, Kwang Man Lee
- Korean J Cytopathol. 2001;12(1):49-52.
- 1,428 View
- 21 Download
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Abstract
PDF
- Salivary duct carcinoma is an uncommon aggressive malignant epithelial neoplasm with similarity to intraductal carcinoma of the breast. This neoplasm occurs most often in the parotid gland of middle-aged and older males. About 7% of reported tumors occured in the submandibular gland. The report of salivary duct carcinoma with calcification is rare. We report a case of salivary duct carcinoma with calcification in the submandibular gland. The patient was a 73-year-old male with a mass of the right submandibular gland for 1 year. On the fine needle aspiration cytology, the aspirate showed scant cellularity, small clusters of tumor cells, and scattered small calcifications. Nuclei of the tumor cells showed mild pleomorphism and round to oval in shape, and cytoplasm was abundant and finely granular. Nucleoli were indistinct and necrosis was not noted. There were no cribriform or papillary arrangements of tumor cells. Cytologic findings of salivary duct carcinoma are variable depending on histologic findings, and calcifications could be an additional cytologic finding.
Original Article
- An Effective Role Pulsed Unipolar Magnetic Field for Bony Decalcification.
- Suk Keum Lee, Eun Young Chung, Gi Jin Kim, Dae Beom Song, Jo Ho Kim, Je G Chi
- Korean J Pathol. 1993;27(2):125-133.
- 1,379 View
- 11 Download
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Abstract
PDF
- To achieve optimal decalcification in tissue and tissue preservation, we have tried magnetic field method and made some promising results. We used pulsed unipolar magnetic field obtained by rectification of 250 V-60 cycle, A.C. As a new method of bony decalcification, using 5% nitric acid, 10% formic acid and 10% formic acid+3% hydrochloric acid solutions, experimental groups were decalcified in the center of the magnetic field. The concentration of calcium ion in the decalcifying solution was measured by calcium-oxalate turbidity test by photometry method, and direct visualization of calcium radiopacity was obtained by soft X-ray view during the decalcification process. The pH change during decalcification was continuously checked and needle penetration method was also used. All the decalcification solution used in this study showed accelerated effect of bony decalcification in the strong magnetic field. Among them 5% nitric acid produced complete decalcification for the medium size bony specimen (less than 10x10x10 mm) within 24 hours, and the histologic feature was almost free of acid-chemical degeneration. The pH of all the decalcification solutions decreased in the strong magnetic field, maximum within 4~6 hours, and kept strong acidity throughout the decalcification procedure. After removal of the magnetic field the pH of all the decalcification solution returned to their original values after 24 hours. It was presumed that the cause of the accelerated decalcification in the magnetic field was due to combined effects of the rapid increase of acidity and the increased molecular resonance to stimulate the ionization of mineral elements.
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