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      Morphologic Analysis of Cytomegalovirus Infected Cells in Bronchial Washing Cytology: Comparison of Liquid-Based Preparation and Conventional Smear

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          Abstract

          Background:

          The cytopathic effects of cytomegalovirus (CMV) infection have been well described since the virus was first reported; however, the morphology of CMV infection has not been clearly studied. We examined the difference in detailed cytologic findings in bronchial washing cytology between liquid-based and conventionally prepared smears.

          Methods:

          Bronchial washing cytology was processed using either the conventional preparation (CP) or liquid-based preparation (LBP). Sixty-nine cells with typical cytopathic effects of CMV infection were detected on CP slides and 18 cells on LBP slides. Using the image analyzer, area, circumference, major axis, and minor axis of the cytoplasm, nucleus, and intranuclear inclusion were measured in singly scattered CMV-infected cells, and histiocytes were used as a control.

          Results:

          The mean cytoplasmic area of CMV-infected cells was 1.47 times larger than that of histiocytes in CP and 2.92 times larger in LBP (p<.05). The mean nuclear area of CMV-infected cells was 2.61 times larger than that of histiocytes in CP and 4.25 times larger in LBP (p<.05). The nucleus to cytoplasm ratio and intranuclear inclusion to cytoplasm ratio of the mean area, circumference, major axis, and minor axis in CP were larger than those in LBP (p<.05).

          Conclusions:

          The sizes of cytoplasm, nucleus, and intranuclear inclusion were larger in LBP than in CP, indicating that CMV-infected cells are easily detectable in LBP. However, the nucleus-to-cytoplasm ratio was larger in CP, suggesting that differentiation from malignancy or regenerative atypia requires caution in CP.

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          Most cited references39

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          Liquid-based preparations versus conventional cytology: specimen adequacy and diagnostic agreement in oral lesions.

          To compare specimen adequacy and diagnostic agreement between liquid-based preparations and conventional smears in oral lesions, and to test the viability of immunocytochemical assay in liquid-based preparations from oral carcinoma lesions.
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            Isolation of intranuclear inclusion producing agents from infants with illnesses resembling cytomegalic inclusion disease.

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              A FILTERABLE VIRUS PRESENT IN THE SUBMAXILLARY GLANDS OF GUINEA PIGS

              In the lesions of herpes simplex and similar conditions due to filterable viruses, cells are present which show characteristic alterations, particularly in the nucleus. The nucleus of these cells contains a mass which stains with acid dyes. Surrounding this mass is a clear space or halo, within which there are large granules staining with basic stains. These cells are little if at all enlarged. In a few human cases, especially in infants, enlarged cells have been found which contain nuclei showing changes similar to those seen in the abnormal cells of herpes simplex. In the ducts of the submaxillary glands of guinea pigs, Jackson observed structures which she considered to be protozoan parasites. Our own studies indicate, however, that these structures are greatly swollen epithelial cells with nuclei having the same characters as the nuclei of the atypical cells in the lesions of herpes simplex. These cells are usually surrounded by a mononuclear cellular reaction. They were found in 84 per cent of the full grown guinea pigs examined but they were present in only three of forty-three young guinea pigs less than 1 month old. The resemblance of these cells, except as regards size, to the atypical cells present in lesions due to filterable viruses suggested that they also may be the result of an infection with a similar agent. That they are usually not present in guinea pigs less than 1 month old indicates that natural infection usually occurs after this period. Experiments were therefore undertaken to determine whether or not an infective agent is concerned in this condition and if so to learn something of its nature. When an emulsion of the submaxillary glands of full grown guinea pigs is injected into the brains of young guinea pigs the animals have fever and exhibit symptoms of cerebral irritation. They usually die in 5 to 7 days and in sections of the brain a diffuse subacute meningitis is found. In the exudate there are large numbers of cells having all the characteristics of the abnormal cells of herpes simplex. Similar cells are present in the lesions resulting from the injection of the same emulsion into the testicle, lung, tongue, and submaxillary glands of young guinea pigs. In none of these lesions, however, are the cells greatly enlarged as they are in the lesions in old guinea pigs. These results support the view that the lesion in the submaxillary gland of old guinea pigs is due to an infective agent. Attempts were therefore made to transmit this agent through a series of young guinea pigs. When the injections were all made into the same organ all the experiments but one gave negative results, but when the site of injection was changed at each transfer it was possible in a number of instances to reproduce the lesions through two animals in series and in one experiment through three animals in series. By modifying the technique, efforts were made to transmit the infection indefinitely but these attempts were unsuccessful. No explanation can be offered for this failure. Studies made to determine some of the properties of the infective agent have shown that it is destroyed by heating at 54° for 1 hour, and that it is not injured by preservation in 50 per cent glycerol for as long as 11 days. After the material had remained in 50 per cent glycerol for 28 days, however, it was found to be no longer infective. The infective agent was not held back by a Berkefeld N filter which was impermeable to bacteria. It seems probable therefore that the infective agent belongs in the group of filterable viruses, though further work will be necessary to learn more of its exact nature. These observations present additional evidence that the presence of cells with nuclear inclusions in any lesion indicates that the injury is probably due to an infective agent belonging in the group of filterable viruses.
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                Author and article information

                Journal
                J Pathol Transl Med
                J Pathol Transl Med
                JPTM
                Journal of Pathology and Translational Medicine
                The Korean Society of Pathologists and the Korean Society for Cytopathology
                2383-7837
                2383-7845
                March 2016
                15 February 2016
                : 50
                : 2
                : 147-154
                Affiliations
                Department of Pathology, Gachon University Gil Medical Center, Incheon, Korea
                Author notes
                Corresponding Author: Seung Yeon Ha, MD, PhD Department of Pathology, Gachon University Gil Medical Center, 21 Namdong-daero 774beon-gil, Namdong-gu, Incheon 21565, Korea Tel: +82-32-460-3073 Fax: +82-32-460-2394 E-mail: syha@ 123456gilhospital.com
                [*]

                Jae Yeon Seok and Jungsuk An contributed equally to this work.

                Article
                jptm-2015-12-25
                10.4132/jptm.2015.12.25
                4804150
                26875760
                ad3a794e-5f74-451e-a473-bfc118239a97
                © 2016 The Korean Society of Pathologists/The Korean Society for Cytopathology

                This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License ( http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted noncommercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

                History
                : 3 November 2015
                : 15 December 2015
                : 24 December 2015
                Categories
                Original Article

                cytomegalovirus,bronchial washing,cytology,lung
                cytomegalovirus, bronchial washing, cytology, lung

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