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\ 436 SAMIMI, ZISKINO, AND WfclLL REFERENCES Allen, G., Samimi, B., Ziskino, M., ano Wlill. H. ((974). X-Ray Diffraction determination of alpha- quartz in respirable and total dust samples from sandblasting operations. Airier. Indust. Hyg. Assoc. / 35,711-717. ' BoeEAfL J.. Hanemann. S.. ano Beven. T. (1962). Silicosis in Louisiana: Ncu or unrecognized industrial hazards. / la. State Med. Sac. 114,391-398. Carlson. A~ and Banks, C. 0952V Spectrophotooteiric determination of siliceous atmospheric contaminations. Anal. Chem. 24,472-476. Drinker. P., ano Hatch. T. (1954). Industrial Dust. 2nd ed. McGraw-Hill. New York. Hunter, D. (1962V The Diseases ofOccupation, 3rd ed. McGraw-Hill. New York. Lipfman. M. (1970). Respirable dust sampling. Amcr. Indust. Hyg, Assoc. / 31, 138-159. Mexewether, E. (1965). Industrial Medicine. Butterworth. London. Patty, A. (1948). Industrial Hygiene and Toxicology. Interscience. New York. Samimi, B. (1973V Silica Dust in Sandblasting Operation. Ph.D. Dissertation. Tulane University, New Orleans, Louisiana. Samimi. B,, Neiuon, A, Weill, H., ano Zismno. M. (1975V The efficiency of protective hoods used by sandblasters to reduce silica dust exposure. A mer. Ind. Hyg. Ass.J. 36, 140-148. Samimi. B-, Weill, H-, and Ziskino. M. (1974). Respirable silica dust exposure of sandblasters and associated workers in steel fabrication yards. Arch. Environ. Health 29,61-66. TALvms, N-. and Hyslop. F. (1958V Colorimetric determination of siliceous atmospheric con taminations. A mer. Indust. Hyg. Assoc. J. 19,54-58. Weidner, R. (1968). Personal Respirable Mass Sampling Procedure. National Center for Urban and Industrial Health, Cincinnati, Ohio. J ECOTOXICOLOGY AND fcNYIRONMLNTAI. SAH TT 1.437--446(1978) al<i_ Some Problems on Health Effects of Polychlorinated Biphenyls Gensyo Umeda, Hiroyukt Nakajima, and Kuniko Yatsuki Kitakyusyu Simitt Kogai Kenkyusyo. Tobata-ku Nakabaru lligasi 3-10-17 Kiiakyushu. P.C. 804. Japan Received June 24.1977 Quantitative and qualitative analyses of polychlorinated biphenyls (PC8) were performed by gas chromatography and the relative pealt height method. Samples for the determination of PCB were obtained from Yusho patients and from normal persons and after necropsy. The relative peak height ratio of PCB in the blood of Yusho patients was applied to differentially diagnose the disease, hut it was difficult to detect the health efTects of PCB taken in through the food chain. The pathological processes of PCB poisoning were not always accompanied by specific signs and developed rather insidiously. The higher chlorinated components of PCB were gradually accumulated into the human organism. It was especially important that PCB crossed thepjacentt^bai^er and was accumulated into the fetus. Th^JetusjvayhejjfigetTor^Rc pfR in the frnfju- The daiaTrom necropsy showed that PC If "cboid also qosi the bloorizh^^BameriFrorn the viewpoint of ontogeny and phytogeny, it is hard to deny tkesenousiniloence uponuwneat generation. In 1968, there occurred, mainly in southern Japan, the so-called Kanemi Yusho Incident. From 14,000 to 15,000 persons were thought to be affected, with polychlorinated biphenyls (PCB) as the causative agent, but only about 1400 persons have been diagnosed as having Yusho disease. As the classification of the disease by Umeda (1972a, 1973a) has shown, the development of the disease ranged widely from typical poisoning to latent or vague health effects. Yusho disease was a typical PCB poisoning that had been caused by consuming food cooked in oil contaminated with Kanechlor 400. the main components of which were tetrachlorobiphenyls. The intoxication was different from the occupational poisoning, which was brought about by exposure to pollutants by inhalation or direct cutaneous contact. Many problems remain which have not yet been resolved practically and theoretically in the medical field, because the Kanemi Yusho Incident was indeed unprecedented in the history of medicine. It was suggested that higher chlorinated biphenyls and their metabolites might still be producing unknown effects in the human organism. We began to study the Yusho disease in 1969 and discovered several fundamental principles in the pathological processes of PCB poisoning and probable relationship between environmental pollution with PCB and its health effects. The fetus is considered to be the target for PCB concentration in the human body, and. from the srandpoint of the health effects of environmental pollutants, we describe in this report (he particular problem of fetal contamination with PCB because this problem is the most serious matter for the coming generation. 437 0147-4513/74/0014-0437 S02.00f0 Copyn*ta'O 1471 toy Ai'aiMm*,-PrTM. If*. Afl n*hu o( reprodw.twn in any fora rewrvai. PnnttU Ui Great fefMua DSW 032498 DEC 0.. 1980 STLCOPCB4016461 \ 438 UMEDA, NAKAJIMA, AND YATSUKI MATERIALS AND METHODS Blood samples were collected by venipuncture from patients and controls, tissue samples were obtained after necropsy or from surgically removed materials, and placentas were supplied by volunteers after spontaneous delivery or stillbirth. PCB was determined by gas chromatography. The analysis was carried out using a Shimazu GC-5A chromatograph equipped with an electron-capture detector fitted with a 2-m by 3-mm coiled glass column packed with 2% SE-30 on Chromosorb W (AWDMCS), 100-120 mesh. The nitrogen flow rate was 50 ml/min and the oven was operated at 200C. The injector and detector temperatures were adjusted at 250 and 210C, respectively. Quantitative analysis was made by peak height measurements. Each peak height in the gas chromatogram was compared with that of peak 16 in Fig. 2, and the relative peak height ratio was calculated by the following formula (Nakajima et al,, 1974a). **i-Phl/Ph,, where R H is the relative height ratio of peak i to peak 16, Ph, is the height of peak i, and Ph,, is the height of peak 16. The lipoperoxide in human blood serum was determined by the thiobarbituric acid (TBA) reaction. The TBA reaction was carried out according to the method of Yamasaki (1972). The absorbance at 535 nm obtained by this method was recorded as the TBA value. The analysis was made on 73 samples from Yusho patients and on 19 samples from normal individuals. RESULTS AND DISCUSSION The clinical picture of Yusho disease (Umeda, 1972a, 1973a) has become extremely complicated, and this is probably due to the effects of higher chlorinated biphenyls and their metabolites in the organism. Some investigations were made on the relationship of the changes in clinical findings to the quantity and quality of PCB in the blood. As indicated in Table 1, the concentrations of blood PCB in Yusho patients were higher than those in controls in 1972 (P < 0.05) and in 1973 (P < 0.05). In 1974, however, there was no significant difference between them. In this way, the TABLE 1 Concentrations of PCB in Human Blood Samples" Yusho patients Controls Mean* + SD Number of samples 1972 13+11* 18 1973 10 + 6* 23 1974 i+3 % 1972 4+2 17 1973 5+2 8 " A 3-year comparison of Yusho pauents with controls. * Mean value of PC B concentrations in pans per billion on a wet weight basts. r The difference from controls is statistically significant. P < 0.05. 1974 4+1 18 HEALTH EFFECTS OF POLYCHLORINATED BIPHENYLS 439 concentrations of blood PCB in Yusho patients have gradually decreased year after year. The relative peak height ratio of PCB isomers in the blood of Yusho patients and controls is shown in Fig. 1. There were differences in the peak height ratio between Yusho patients and controls, that is, the relative height ratio of peak 14 to peak 16 in thepatients was lower than that in controls, but the ratio of peak 21 in the patients was Fig. I. Relative peak height ratios of PCB in human blood samples. Relative peak height ratio of various peaks is in tdation to peak 16 1 1.00. Comparison of Yusho patients with controls for 3 years. #. 1972; a 1973; A. 1974. higher than that in controls. These differences have contributed io diagnosing Yusho disease. Epidemiological surveys in the Goto islands were made in 1972 to establish the relationship between environmental pollution with PCB and its effects upon the inhabitants. As indicated in Table 2, the concentrations of PCB in marine organisms were higher than those in human blood. The gas chromatograms of PCB in marine products and human blood are shown in Fig- 2. The relative height ratio of peak 14 to peak 16 in human blood was higher than that in marine organisms (visceral organs of OSW 032499 440 UMEDA, NAKAJIMA, AND YATSUKI Sepioleathis Iessoniana). As discussed later on the basis of analyzing human visceral organs (Fig. 4), it was suggested that the lower chlorinated biphenyls might be easily . carried out of the body. The relative increase in higher chlorinated isomers in marine animals was extremely important in the light of health effects of pollutants through the food chain. TABLE 2 . Concentrations of PCB rN Human Blood Samples and in Marine Organisms Type of sample Human blood Tamanoura Controls Possible cases* Probable eases* Naru Possible cases* Probable cases* Classical cases* Marine organisms Carartx equula Muscles Visceral organs Trackurus japonicus Muscles Visceral organs Sphyraena japonica Muscles Visceral organs Sepioteathis Iessoniana Muscles Visceral organs Number of samples PCB 19 S 2 19 7 3 n 8+4 20 6 + 4 4 5 + 2 19 12 + 5 1 130 1 100 1 no l no no 1 420 1 150 1 160 Epidemiological su/veys were performed in two regions. Tsretnoura and Naru, in the Goto Islands in 1972. Values of PCB concentration are means * SO in parts per billion on a wet weight basis. * Of Yusho disease. In this regard, a study was done on PCB contamination of the placenta and umbilical cord blood obtained from controls. As shown in Table 3, the concentrations of PCB in the various parts of the placenta were higher than those in the umbilical cord and in the blood. This fact showed that the accumulation of PCB into the placenta of controls could reflect the general condition of environmental pollution in Japan. Simultaneously, we were able to indicate a very important fact, that PCB does concentrate in humans, particularly in the fetus. Table 4 shows the concentrations of PCB in various tissues obtained after necropsy of a stillborn baby bom to the Y usho mother. PCB was accumulated specifically into HEALTH EFFECTS or POLYCHLORINATED BIPHENYLS 441 Fig. 2. Gas chromatograms of PCB in marine organisms and human blood samples. A. authentic sample (PCB. 300:400:500:600 = l: 1:1:1); B. marine organism {Sepioieaihis lessonianak C. human blood (one of the habitants}. the subcutaneous and adipose tissues of humans. The same results were obtained in normal adults (Nakajima et a!., 1974b), and the amount of PCB in them could indicate the environmental contamination as well. Figure 4A shows a gas chromatogram of PCB in the liver of a control stillborn baby and Fig. 4B shows PCB in the liver of a stillborn baby with typical PCB poisoning born in 1968 to a mother with Yusho disease. Figures 4C and 4D show gas chromatograms of PCB in tissue obtained from a stillborn baby born in 1975 to the mother of the baby in Fig. 4B. This baby had no specific signs of PCB poisoning, such as the characteristic dark brownish skin coloration. Figure 4B shows many lower chlorinated components which resemble the causative material of Yusho disease. TABLE 3 Concentrations of PCB in Several Control Placental Parts Samples PCB" Placental villi Umbilical cord Chorion and amnion Umbilical cord blood II + 5 3+ l 10 + 9 4 +3 ' Means t SD of convemralions of live determinations in parts per billion on a .cl weight basis. DSW 032500 442 UMEDA, NAKAJI.MA. AND YAISUKI On the other hand, most of the lower chlorinated components in Fig. 4C decreased, and higher chlorinated components relatively increased. As indicated in Fig. 4. many higher chlorinated components were detected in various parts of the brain of these babies. PCB was detected in the brains of adult Yusho patients as well (Nakajima el a!., 1975), and PCB diffusely infiltrated into the brain cortex and subcorneal structure. The relationship between the phases of poisoning in mother and newborn baby is shown in Table 5. The physical signs in newborn babies did not always correspond to the phases of the disease in the mothers. Adult Yusho patients have developed many complicated symptoms such as general tiredness, disturbance of recent memory, and drastic changes in behavior during the table 4 Concentrations of PCB in Various Tissues of a Stillborn Baby' Sample PCB* Kidney Right Left Spleen Liver Bone marrow Bile Blood Brain Temporal and occipital lobes Frontal lobe Limbic system Subcutaneous adipose tissues Abdominal wall Gluteal region 30 23 15 49 3 13 i 9 10 10 806 683 * Born in 1975 to a mother with Yusho disease. * Concentrations of PCB in parts per billion on a wet weight bails. past few years (Umeda. 1973b; Umeda and Takeuchi. 1973; Umeda ei al., 1975), all of which are characteristic aspects of the slow phase of the pathological processes of Yusho disease (Umeda, 1973c). To make the process of these general nonspecific reac tions clear, we investigated lipoperoxide in blood serum of Yusho patients and controls. As shown in Fig. 5, a significant increase in scrum lipoperoxide was obtained as the TBA value in the patients. Most of the Yusho patients have complained of nervous symptoms, and PCB was detected in the brain. However, we never identified them with any change in the electroencephalograms of Yusho patients (Umeda, 1973d). Furthermore, it was deduced on the basis of the clinical observation and of the examination of transient ischemic attack (Umeda, 1972b) that PCB brought about some change in the central nervous system. To resolve this problem, we administered the Uchida-Kraepelin psychodiagnostic test to the patients and controls. The study was conducted on 45 HEALTH EFFECT'S OF POLYCHLORINATED BIPHENYLS 443 Yusho patients and 69 normals. The rate of the rest effect was calculated by the following formula. R ~ (m Jm,) x 100. where R is the rale of the rest cfTcci and m, and in, are the scores before and after the rest. The distribution of the rale of the rest effect is shown in Fig. 6. The rate of the rest effect in (he patients was significantly lower than that in controls. It became Fic. 3. Gas chromatograms of PCB in several parts of placenta and others obtained from controls. A. authentic sample (PCB, 300:400:500:600 = 1:1:1:1); B, piacenui villi; C, chorion and amnion; 0, umbilical cord blood. obvious that PCB brought about the functional disorders in the higher pans of the centra] nervous system, especially the cerebral cortex. From the viewpoint of onto genesis and phylogenesis, this is a serious matter, because the brain cortex of the fetus is less resistant to harmful materials such as PCB. SUMMARY We reported clinical studies on Yusho disease and epidemiological surveys on the contamination with PCB of humans and of the environment: (1) Yusho disease is caused by (he ingestion of contaminated cooking oil with PCB, and the clinical picture changes into a more complicated one after the onset of the disease. On the basis of studies on Yusho disease, it has been shown that it is very OSW 032501 STLCOPCB4016464 444 UMEDA, NAKAJIMA. AND YATSUKI $ 10 20 30 40 SO sin. Fc. 4- Gu chromatograms of PCB in a stillborn baby. A. liver (control, 1968); 8. liver (bom to a manifest-type mother with Yusho disease. 1968); C. liver (bom to the same mother as in 8. 1975); D. limbic system (the same case as in C. 1975; other parts of the brain showed the same pattern as in D in the gas chromatogram). A and 8 were provided by Professor M. Kikuchi, Fukuoka University School of Medicine. ' TABLE 5 Relationship between Poisoning Phases in Mother and Newborn Baby Case* Mother, phase of disease Newborn baby, resulted in . 1 Latent, free of symptoms Typical manifest* 2 Manifest, mild Atypical, free of symptoms 3 Manifest, moderate Manifest, mild 4 Manifest, advanced Typical manifest* 5 Manifest, poor in symptoms i Fetuses aborted, the same levels 6 Manifest, advanced j of PCB concentrations 4 1. mother was of latent-type poisoning and had no symptoms, but delivered a typical manifest type; 2-4. newborn babies reflected well the seventy of the disease in the mothers, ranging from atypical to typical poisoning; 5 and 6, PCB concentrations in both fetuses (3 and 4 months old) were at the same levels independently of the symptoms and signs ui the mothers. Characteristicdark brownish skin coloration. HEALTH EFFECTS OF POLYCHLORINATED BIPHENYLS 445 j* r r J. Controls Patients Fig. 5. Lipopcroxide levels as ihc TBA value of blood serum of Yusho patients and controls. Means : SD: Yusho patients. 0.025 0.007 (n =. 73); controls. 0.014 + 0.004 (n 19). difficult to detect clearly the health effects of PCB taken in through the food chain because PCB intoxication is not always accompanied by specific physical signs, as the classification of Yusho disease by Umeda had indicated. (2) Quantitative and qualitative analyses of PCB in the blood were done to diagnose Yusho disease, particularly its visceral and latent types of poisoning, whiefuare l------------------- 1--------------------1------------- * M I . ~ : ' Ra no 8 gftOOO no 0 90 Agtf P. . C. D P So 8000 F .1 8 ?. c. 0 So 8goo gooao t 0 p. c. air.. o oo L l oo ooo * So J 4 r. c. ;.r. f, i 808080000 888oo.YOOuO.) ooooOoooojo 8* M* 8 f\ c. Fofj 1 Mtif III 0 117.j 11:. o i: i. i iij.4 1:1.4 m i i; i i i.i i :o ; rSt 4 :7.9 !7.S .29.: s.G :7 i 9.5 *9 4 - ij 1 18 .S J3 1 1 IN -> SL o.<>: i*<o o5 o o:*f-u.ui 1 dill- P-0 Ol Fio. 6. Distribution of the race of rest effect. P. patterns, C. controls; SL. significant level. R * x 100. where R is the rale of rest effect and m, and m, are (he scores before and after the rest. DSW 032502 446 UMEPA, NAK.AJ1MA, AND YATSUK1 composed mainly of nonspecific pathological processes. However, to analyze the pathophysiological changes in (lie human organism as a whole, further examinations are needed, for example, of Upoperoxide as the TBA value in blood serum and in other organs. (3) The fetus is considered the target for PCB concentration in humans because PCB induced fetopathy, and the data obtained after necropsy have indicated that PCB crossed the placental barrier and accumulated into the fetus. . (4) Simultaneously, it was very important that PCB was detected in the brains of adults and stillbirth infants, because this makes It clear that PCB could also cross the blood-brain barrier. The cerebral cortex is the youngest part of the central nervous system and, from the standpoint of ontogenesis and phylogenesis, is less resistant to hazardous agents. Therefore, it is hard to deny the serious influence of PCB on the next generation. REFERENCES Nakajima. H-, Yatsukl K,, and Umeda. G. (1974a). Polychlorinated biphenyls in human WoodComparison of ?C8s intoxicated cases with normal control subjects, Med. Biol. 89,3-9. Nakahma, H., Yatsukz, K., and Umeda. G. (1974b). Determination of PCB in human blood and tissues. 47th Cong. Japan fndusi. Hyg. 258-759- Najajima. H-, YArsuxu K-. and Umeda, G. (1975). Analysis of gas chromatogram patterns of PCB in various tissues obtained from necropsy of Yusho patient (preliminary report). Japan. J. Indust. Health. io press. Umeda. G. (1972a). PCB poisoning in Japan. AMBlO 4,132-134. Umeda. G. (1972b). TlA-like symptom in PCB poisoning. Japan. J. Indust. Health 14, 53--54. Umeda. G. (1973a). On problems around poisoning from PCB./. Japan, Sci. ft, 19-25. Umeda, G. (1973b). Clinical study on PCB poisoning. Rodo no Kagaku 28,36--42. Umeda, G. (1973c). Symptoms of PCB poisoning. 1st Hat Symp. HESC. abstr.. 59-61. Umeda. G. (1973d). Clinical aspects of PCB poisoning (continued). 46th Congr. Japan fndusi. Hyg. 358-- 359. . Umeda. G_ Nakajima, H-. and Yatsuki. K. (1975). Some problems on clinical picture of Yusho as PCB poisoning. Japan. J. Hyg. 30,133. Umeda. G.. and Takeuchl M. (1973). Pathophysiology of higher nervous activity in PCB poisoning. Japan. J. Brain Physiol. 113,126. Yamasaki. S. (1972). Quantitative determination of lipoperoxide. Japan. J. Clin. Pathol. Special Issue No. 19, 128-140. ECOTOXICOl.OOY AND ENVIRONMENTAL SAFETY 1,447-455 (1978) Radionuclide Movement in an Ecological Chain B. Glowiak ano J. Pacyna Environment Protection Engineering Institute. Technical University of Wroclaw, ui. Wybreze Stanislava Wyspianskiego 27, JO-370 Wroclaw, Poland Received May 12.1977 Radioactive substances create a considerable hazard, is they move through the environment of man and form an ecological chain. The first link of this chain is atmosphere; the next links are formed by water, soil, plants and animals. At the end cf the chain is man. The present paper provides a model of strontium-90 and cesium-137 transfer from the atmosphere to man over Wroclaw (Poland) and its vicinity. It was found that air contamination is responsible for contamination of the remaining media. This correlation is shown in terms of mathematical formulas. Strontium-90 determinations were made in samples by using radiochemical methods, whole Ca-137 was determined by spectrometry. The apparatus employed was made in Poland. Our testing methods and apparatus have gained a high mark in the Intercomparison Runs sponsored by IAEA. Radioactive substances create a special form of contamination. The noxiousness of those substances considered to be a hazard for the natural environment is well known. However, the development of nuclear engineering, as well as nuclear weapons tests, aad the search for new and cheap power sources have considerably increased the concentration of radionuclides in particular environment media. Radioactive con tamination is known to move through the environment and form an ecological chain. The first link of the chain from which radionuclides begin to move is atmosphere. The subsequent links are formed by fallout, soil, water, plants, and animals, respectively. At the end of the chain is man, constituting the last link in the system. Of the many radionuclides present in the atmosphere, stromium-90 and cesium-137 require special attention. These radionuclides are characterized by a long half-life period. That is why we have chosen to use these when carrying out investigations intended to give an idea ofthe movement of radioactive pollution in an ecological chain. Studies and measurements concerned with this problem were performed at the Environment Protection Engineering Institute of the Technical University of Wroclaw (Poland) during the last $ years. Measurements were carried out continuously in the city centre of Wroclaw (Glowiak and Pacyna, 1974a). Results obtained from investigating the *Sr and l,TCs concentrations in the atmosphere during the recent 3 years are shown in Fig. 1. An analysis of those data indicates the appearance of a maximum concentration in the spring months. This concentration stems from increased coal combustion during the winter heating period when larger amounts of pollutants are emitted into the atmosphere. K similar maximum concentration is observed when investigating fallout and water radioactivities. The results of fallout measurements are shown in Fig. 2. 447 OH7-4JH/78/QOU-04<7SO.'00 0 As-kJcrn*, Prt,, All npH> of ,n fiMitt rcMrvcU FruiicU m G'cai Bniiin DSW 032503