Document 3Nn9JM9EBnQaJ0kBjqY2xdr83

Bull. Environ. Contam. Toxicol. (1993) 50:325-332 1993 Springer-Verlag New York Inc. 'Environmental I Contamination land Tbxieology Asbestos Body and Fiber Concentrations in Pathological Autopsy Tissues of Patients with Malignant Peritoneal Mesothelioma Katsumi Saitoh,*1 Hajime Muto,2 Noriyuki Hachiya,3 and Yukio Takizawa3 'Division of Environmental Science, Akita Prefectural Institute for Fisheries and Fisheries Management, Unosaki 16, Daishima, Funagawakou, Oga-shi, Akita 010-05, Japan; ^Environmental Research Center, Akita University, Akita 010, Japan, and department of Public Health, Akita University School of Medicine, Akita 010, Japan It has long been known that diffuse interstitial pulmo nary fiberosis can be caused by asbestos(Doll and Petro 1985). Definite exposure reponse relations between both level and duration of exposure to asbestos and presence of definite radiographic abnormalities have been shown by many authors(Finkelstein and Vingilis 1984; Copes et al. 1985). Asbestos bodies in lung tissue have been re cognized as a marker of past asbestos exposure (Churg and Warnock 1977). Although there is no convincing evi dence that indirect exposure contributes to the occur rence of mesotheliomas, there have been numerous re ports of this rare tumour in dividuals exposed to as bestos (Anderson et al. 1976; Vianna and Polan 1978). From the review of all cases newly diagnosed in 1982 as a malignant mesothelioma of the pleura or peritoneum (Churg 1985), it has been shown that the incidence rate of mesothelioma in British Columbia has increased near ly six times for men compared to the period 1969 to 1975, but remained roughly unchanged for women, and al most all of the cases in men in this series could be linked to asbestos exposure, de Klerk et al.(1989) have predicted future incidence of asbestos-related disease in former Wittenoom asbestos workers in Western Austra lia for the period 1987 to 2020. They predicated 2898 deaths in this period, 692 cases of mesothelioma, 183 cases of lung cancer, and 482 cases of asbestosis. Ad ditionally, they indicated that the incidence of both lung cancer and asbestosis was greatest in those sub jects with the highest levels of exposure to crocidolite and in smokers(de Klerk et al. 1991). Berry(1991) Send reprint request to Katsumi Saitoh. 325 also predicted from the follow-up study on Wittenoom workers that between 250 and 500 deaths and between 340 and 465 deaths will occur due to mesothelioma and lung cancer, respectively. It is important for mesothelioma to have criteria other than history by which a case may be classified as as bestos-related (Warnock 1989). The risk of malignant mesothelioma associated with low-level asbestos expo sure is an important unresolved issue today(Mowe et al. 1985). We report here a case study on malignant perito neal mesothelioma associated with asbestos, using a scanning electron microscope with x-ray microanalyzer and a phase-contrast microscope. MATERIALS AND METHODS The source population for both cases and controls was the population of Akita city in the North part of Japan in 1987. The incidence rate per year of carcinomatous peritonitis including malignant peritoneal mesothelioma in Japan has been reported as about 3.3 per million for general population from 1985 to 1989(Ministry of Health and Welfare, Japan 1985-1989). Two patients diagnosed as malignant peritoneal mesothelioma in Akita Kumiai General Hospital were subjected, and three populations who died of myocardial infarction or dissecting aortic aneurysm in the hospital were chosen as controls. Table 1 shows their profiles. Analytical procedure used to determine asbestos bodies and fibers in pathological autopsy samples has been described elsewhere(Ashcroft and Heppleston 1973).About 5 g of the sample was homogenized with distilled water at high speed in a Ultrahomogenizer(Physcotron: Ikemoto Sci. Technol. Co., Japan). An aliquot of the sample was transferred to a centrifugal tube of 50 ml equipped with a condenser and saponified with 20 ml of 40 % KOHethanol solution in water bath at 100C for an hour. After cooling, the sample was filtrated through a mem brane filter(millipore, AA 0.8^u mx47^mm) with 150 ml of distilled water. Asbestos sample trapped on filter was kept in a desicator for the determination using a phase-contrast microscope(Olympus, BH- I , Japan) and a 326 Table 1. A profile of cases and controls Case/Control Age/Sex Smoking Occupation Cause of Death Case 1 2 62/M 42 /F none none coachbuilder(+) nurse)-) malignant peritoneal mesothelioma malignant peritoneal mesothelioma 1 Control 2 3 63/M 79/F 76/F 9 pk/yr none none painter(+?) housewife(-) housewife(-) myocardial myocardial dissecting aneurysm infarction infarction aortic Note. Probability of occupational asbestos exposure: probable(+), possible(+?), unlikely or unknown(-). scanning electron microscope (Hitachi S-7000, Japan) equipped with an energy-dispersive x-ray microanalyzer (Kevex, Deltea HI, U.S.A). Prior to the determination, asbestos sample was prepared by the acetone-triacetin method reported elsewhere (Japan Asbestos Assoc. 1988). The dry weight of sample was equal to one-half of wet one. Chemicals were of reagent quality and obtained from Wako Pure Chemical Industries (Japan). RESULTS AND DISCUSSION Asbestos body and fiber concentrations in lung, greater omentum, and large intestine tissues of cases and con trols are shown in Table 2. For 62-year-old male of Table 2. Concentrations of asbestos bodies and naked fibers in lung, greater omentum, and large intestinal tissue samples from two patients with malignant peri toneal mesothelioma and three general populations (per g dry base) Case Control Age/Sex 62/male 42/female 63/male 79/female 76/female Tissue/Sample ABa NF^ AB NF AB NF AB NF AB NF o 1 Is Lung Nontumor 20-28 36-48* NAC NA 20-28 12 4 4 ND-4 Greater Tumor 12-44* 20-36* ND-4 8-12 NA cmentum Nontumor ND-12 24-72* ND ND ND NA NA NA NA NA ND-12 ND-4 8-12 ND ND Larqe Tumor intestine Nontumor NA NA NA ND ND NA NA NA NA NA NA NA ND ND ND-16 ND-4 ND-4 ND ND ND Note. Three samples per tissue were prepared. The ranges shown in the table are data from three observations. aAsbestos body, bNaked fiber, cNot analyzed,dNot detected, * p<0.05 by the Mann- Whitney U Test. 327 case 1, asbestos body concentrations in lung tissues ranged from 20 to 28 per g dry base and corresponded to the lev el which Zhang(1987) has classified to the slight exposure level by asbes tos body as the ranges of 11 to 100 per g wet lung. Asbestos body size of 100 n m at the maxi mum was observed. The naked fiber concentra tions in lung tissues were approximately two times, compared to those of asbestos bodies. For greater omentum, the bodies and fibers were Figure 1. A photograph of asbes tos body (bell shape type) in greater omentum sample of case 1. also found in tumor or nontumor tissues. Their concentrations in tomor tissues were similar to the lung tissue levels, and their sizes were about 20 yum for body and a few hundred yum for fiber. Most of body types were bell and club shape types. A phase-con trast microscopic photograph of asbestos body in great er omentum tissue of case 1 is shown in Figure 1. Fiber concentrations for lung and greater omentum of case 1 were significant(p<0.05) as compared with controls, us ing the Mann-Whitney U Tests. Furthermore, chrysotile fibers were identified in lung tissues of case 1, using the scanning electron microscope with energy-dispersive x-ray microanalyzer (see Figures 2 and 3). However, in order to the small number of patients and the lack of specimens of greater omentum and large intestinal tumor samples in the control patients, it is not clear whe ther or not the differences in fiber concentrations be tween case 1 and controls are shown. Fiber size is believed to play a role in determining risk of a particular asbestos-related disease (Lippmann 1988). Warnock(1989) has quantified the lung asbestos 328 NBS348-20kv Vert" 2000 counts Dlsp* 1 Si 9 Figure 2. A photograph of chrysotile fiber in lung sample of case 1, using an electron microscope. <- 0.000 Range- 10.230 keV Figure 3. X-ray diffraction spectra for lung sample of case 1, using an electron microscope with x-ray micro analyzer. burden in shipyard and construction workers with meso thelioma, and reported that their burden was signifi cantly greater than the burden found in men of general populations(p<0.001). Furthermore, because that the median concentration for total amphibole fibers in sub jects with mesothelioma did not differ significantlly as compared with subjects with asbestosis, it was hypo thesized that fiber size, especially amosite of the most prevalent type, would differ among asbestos-re lated disease. For our case study, it was suggested that the signifi cant difference in fiber concentrations in greater omentum tissues of a patient (case 1) with malignant peritoneal mesothelioma would associate with asbestos exposure. Then, the pleural plaques were not observed from pathological findings of case 1. However, for the relation between asbestos fiber types and pleural plaques in a general autopsy population, it has been suggested that the presence of pleural plaques corre lates with a modest(50-fold) increase in numbers of long high-aspect ratio commercial amphiboles (amosite and crocidolite) in lung tissue (Churg 1982). For 42- 329 NBS348-20kv Vert* 2778 counts Dlsp* 1 Si Figure 4. A photograph of sil ica fiber in greater omentum sample of case 2, using an electron microscope. Figure 5. X-ray diffraction spectra for greater omentum sample of case 2, using an electron microscope with xray microanalyzer. year-old female of case 2, although a few numbers of asbestos bodies and fibers were detected in tumor tis sues of greater omentum, their concentrations were much low, and the body types were similar to case 1. How ever, they were not observed in nontumor tissues of greater omentum and in tumor and nontumor tissues of large intestine. On the other hand, a large quantity of fiberous substances of needle-type were observed in greater omentum tissues. It was found that the materi als were silica fibers (see Figures 4 and 5), using the scanning electron microscope. Silica fibers were not in above tissues of case 1 and controls. Considering that mesothelioma is also induced by the stable and fiberous substances other than asbestos in vivo (Stanton et al. 1981), it was suggested that asbestos as well as fiber ous substances such as silica fibers might play an important role to malignant peritoneal mesothelioma. 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