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Reply to Drs. Perkins and Cauchi
CORRESPONDENCE
Hydatoxi tuatba -An artifact
To the Editors: A great deal of interest has been generated by the report of Lueck and associates (AM. J. 0BSTET. GYNEcoL. 145:15, 1983) that an organism (Hydatoxi lualba) was detected in patients with gestational trophoblastic disease and in patients with "toxemia of pregnancy." We endeavored to repeat these findings and have, indeed, found organisms as described by Lueck and associates. We submit that these organisms are an artifact for the following reasons. 1. They were present in all samples of blood examined irrespective of origin. 2. We could reproduce these artifacts by the addition of debris, such as fibers from Kleenex tissues and cotton wool, to the blood prior to spreading the blood film. 3. In plasma from normal subjects which had been filtered through a 0.22 ~m Millipore filter, no organisms were detected. However, if fibers (as described in item 2) were then added to this plasma, numerous "'organisms" could be seen. 4. Using these techniques, we could observe the socalled "larva-like" and "egg-like" forms, as well as the "worm-like" forms. (Photographs of these "organisms" are available on request.) We submit that these "organisms" were artifacts that resulted from debris that was introduced into the blood at any time from collection until spreading onto a slide. Onto this backbone of debris, a coagulum of plasma proteins was deposited during the spreading and drying of the blood. The harsh method of staining used, involving ether-sulfuric sulfation, will expose alcohol groups which will stain metachromatically with the toluidine blue staining procedure. Diane Perkins M. N. Cauchi Department of Pathology The Royal Women's Hospital 132 Grattan Street Carlton. Victoria, 3053, Australia
Reply to Drs. Perkins and Cauchi To the Editors: Drs. Perkins and Cauchi have reproduced artifacts similar to three of the four forms described by us ("worm-like," "larva-like," and "egg-like"), by contaminating blood samples with fibers from Kleenex tissue and cotton wool. We also reproduced such artifacts, as
stated in the article. We hope to have the opportunity to review their material and compare it with ours before we can comment any further. It is important to point out that Drs. Perkins and Cauchi have not reproduced the "sperm-like" structure. Neither did we. judith Lueck John I. Brewer, M.D., Ph.D. Silvio Aladjem, M.D. Marilyn Novotny, M.S. Loyola Stritch School of Medicine 2160 South First Avenue Maywood, Illinois 60153 Northwestern University School of Medicine 303 East Chicago Avenue Chicago, Illinois 60611
Vermiform structures in blood in gestational trophoblastic disease To the Editors: A recent report by Dr. Lueck and associates (AM. J. OBSTET. GYNECOL. 145:15, 1983) described vermiform structures in peripheral blood of patients with preeclampsia, eclampsia, and gestational trophoblastic disease. The authors observed morphologic similarities between these structures and several orders of helminths; the authors considered these structures to be an organism, which they tentatively labeled Hydatoxi lualba. In a companion report (AM. J. OBSTET. GYNECOL. 145:27, 1983), the authors ascribed an etiologic role for this organism in the development of a "toxemia-like" syndrome in beagles. These "worm-like" structures do not appear to be helminths. Several lines of evidence support this conclusion. First, the structures are markedly pleomorphic. Such pleomorphism for an organism is without precedent in parasitology. Moreover, the evidence is insufficient to suggest that the various structures are related to each other. Second, the staining properties of these structures are atypical for helminths. Although Giemsa stain (which stains DNA, RNA, and protein) is especially useful for staining metazoan parasites, only the "sperm-like" structures described by Dr. Lueck and associates stained with Giemsa stain. Since toluidine blue 0- will metachromatically stain complex carbohydrates, 1 the structures stained red in Figs. 1 and 6, A of the first report appear to be composed of carbohydrates, rather than protein and nucleic acid, as would be expected of a helminth.
469
Hydatoxi tuatba -An artifact
To the Editors: A great deal of interest has been generated by the report of Lueck and associates (AM. J. 0BSTET. GYNEcoL. 145:15, 1983) that an organism (Hydatoxi lualba) was detected in patients with gestational trophoblastic disease and in patients with "toxemia of pregnancy." We endeavored to repeat these findings and have, indeed, found organisms as described by Lueck and associates. We submit that these organisms are an artifact for the following reasons. 1. They were present in all samples of blood examined irrespective of origin. 2. We could reproduce these artifacts by the addition of debris, such as fibers from Kleenex tissues and cotton wool, to the blood prior to spreading the blood film. 3. In plasma from normal subjects which had been filtered through a 0.22 ~m Millipore filter, no organisms were detected. However, if fibers (as described in item 2) were then added to this plasma, numerous "'organisms" could be seen. 4. Using these techniques, we could observe the socalled "larva-like" and "egg-like" forms, as well as the "worm-like" forms. (Photographs of these "organisms" are available on request.) We submit that these "organisms" were artifacts that resulted from debris that was introduced into the blood at any time from collection until spreading onto a slide. Onto this backbone of debris, a coagulum of plasma proteins was deposited during the spreading and drying of the blood. The harsh method of staining used, involving ether-sulfuric sulfation, will expose alcohol groups which will stain metachromatically with the toluidine blue staining procedure. Diane Perkins M. N. Cauchi Department of Pathology The Royal Women's Hospital 132 Grattan Street Carlton. Victoria, 3053, Australia
Reply to Drs. Perkins and Cauchi To the Editors: Drs. Perkins and Cauchi have reproduced artifacts similar to three of the four forms described by us ("worm-like," "larva-like," and "egg-like"), by contaminating blood samples with fibers from Kleenex tissue and cotton wool. We also reproduced such artifacts, as
stated in the article. We hope to have the opportunity to review their material and compare it with ours before we can comment any further. It is important to point out that Drs. Perkins and Cauchi have not reproduced the "sperm-like" structure. Neither did we. judith Lueck John I. Brewer, M.D., Ph.D. Silvio Aladjem, M.D. Marilyn Novotny, M.S. Loyola Stritch School of Medicine 2160 South First Avenue Maywood, Illinois 60153 Northwestern University School of Medicine 303 East Chicago Avenue Chicago, Illinois 60611
Vermiform structures in blood in gestational trophoblastic disease To the Editors: A recent report by Dr. Lueck and associates (AM. J. OBSTET. GYNECOL. 145:15, 1983) described vermiform structures in peripheral blood of patients with preeclampsia, eclampsia, and gestational trophoblastic disease. The authors observed morphologic similarities between these structures and several orders of helminths; the authors considered these structures to be an organism, which they tentatively labeled Hydatoxi lualba. In a companion report (AM. J. OBSTET. GYNECOL. 145:27, 1983), the authors ascribed an etiologic role for this organism in the development of a "toxemia-like" syndrome in beagles. These "worm-like" structures do not appear to be helminths. Several lines of evidence support this conclusion. First, the structures are markedly pleomorphic. Such pleomorphism for an organism is without precedent in parasitology. Moreover, the evidence is insufficient to suggest that the various structures are related to each other. Second, the staining properties of these structures are atypical for helminths. Although Giemsa stain (which stains DNA, RNA, and protein) is especially useful for staining metazoan parasites, only the "sperm-like" structures described by Dr. Lueck and associates stained with Giemsa stain. Since toluidine blue 0- will metachromatically stain complex carbohydrates, 1 the structures stained red in Figs. 1 and 6, A of the first report appear to be composed of carbohydrates, rather than protein and nucleic acid, as would be expected of a helminth.
469