Gene polymorphism and frequencies of the NPC1L1 gene (rs2072183, rs217434 and rs217428) in Japanese patients with dyslipidemia.

WHAT IS KNOWN AND OBJECTIVE: Niemann-Pick C1-Like 1 (NPC1L1) plays a pivotal role in intestinal cholesterol absorption. Ezetimibe is known as an inhibitor for NPC1L1 and decreases concentration of low-density lipoprotein cholesterol (LDL-C) in blood. Responses of the decrease of serum LDL-C levels to ezetimibe have been reported to be different among NPC1L1 variants. However, there are still limited data concerning the genetic variation in the NPC1L1 gene, specifically, in Japanese patients with dyslipidemia. The purpose of this study is to elucidate genotype and allele frequencies of the NPC1L1 gene in Japanese patients with dyslipidemia. METHODS: Written informed consent was obtained from all participants. All patients were administered ezetimibe at the dose of 10 mg for once a day either alone or coadministered with statins. Patient's data were retrospectively obtained from their medical records. Genomic DNA was extracted from whole blood samples and analysed three NPC1L1 SNPs (rs2072183, rs217428 and rs217434) by the direct sequencing method. RESULTS AND DISCUSSION: We found that there is a significant difference of genotype frequencies between healthy Japanese and dyslipidemic subjects in rs2072183. No significant differences were observed in rs217428 and rs217434; however, comparison of our data with literature reports suggests that there are significant differences in the frequencies of rs217428 and rs217434 between Canadian and Japanese dyslipidemic patients. WHAT IS NEW AND CONCLUSION: Our study is the first report concerning the genotype and allele frequencies of the gene coding for NPC1L1 in Japanese patients with dyslipidemia. The most notable result was to demonstrate that there exists a significant difference in rs2072183 variant between healthy Japanese and dyslipidemic subjects and also found that there exists genetic variation of rs2072183 between Japanese and Canadian patients with dyslipidemia. Our results are expected to facilitate research in the proper use of ezetimibe-based mono- or combination therapies. Further studies will be required to evaluate the effects of rs2072183 on the efficacy of LDL cholesterol reduction by ezetimibe.

Effect of interleukins response to ECM-induced acquisition of drug resistance in MCF-7 cells.

AIM: To examine the effect of various components of extracellular matrix (ECM) on acquisition of drug resistance to taxol and camptothecin by breast carcinoma cell line MCF-7. METHODS: Cancer cells were cultured on bovine serum albumin (BSA), vitronectin (VN), fibronectin (FN), collagen type I (COL-I), or Matrigel-coated plates with or without taxol (paclitaxel) or camptothecin treatment. The effect of anticancer drugs on cell growth was accessed by XTT assay, and the alterations of cellular morphology were examined by phase contrast microscopy. Immunofluorescence study was performed using monoclonal anti-beta-tubulin antibody. RESULTS: All cell lines showed a significant decrease in cell survival when treated with anticancer drugs without components of ECM, whereas survival rates of Caco-2, MCF-7 and NCI-H292 were significantly increased when cells were cultured on COL-I- and Matrigel-coated dishes after treatment with paclitaxel or camptothecin. MCF-7 cells showed and maintained a colony formation when cultured on the COL-I- and Matrigel-coated dish. Moreover, cytotoxicity (IC50) was decreased by taxol (paclitaxel) or camptothecin treatment during colony formation in MCF-7 cells, suggesting that morphological changes could increase survival of cells treated with anticancer drugs. Thick circumferential bundles of microtubules around the periphery of the cells and chromatin condensation was not observed for MCF-7 cells on COL-I- and Matrigel-coated dishes treated with paclitaxel. To confirm this, spheroid cells were prepared, and we found that cytotoxicity was decreased for these cells, and significantly increased when cells were co-cultured on Matrigel- or COL-I-coated upper wells. The effect of anticancer drugs on cell survival was efficiently inhibited by interleukin-6 (IL-6) and interleukin-8 (IL-8). CONCLUSIONS: Present results suggested that not only integrin-ECM interactions but also other factors such as IL-6and IL-8secreted by cancer cells, cultured on COL-I and Matrigel dishes, are involved in the acquisition of drug resistance by MCF-7.

Dendriform pulmonary ossification in patient with rare earth pneumoconiosis.

The case history is presented of a patient with pathologically proven dendriform pulmonary ossification and rare earth pneumoconiosis confirmed by analytical transmission electron microscopy. This is thought to be the first report of this association.

Airborne fiber concentration and size distribution of mineral fibers in area with serpentinite outcrops in Aichi prefecture, Japan.

Airborne fiber concentrations and size distributions of both asbestos and nonasbestos fibers were determined at property boundaries of 4 serpentinite quarries producing crushed stone (quarry property boundary), 10 sites within 10 km of the area with serpentinite outcrops (serpentinite area), and 2 sites in a reference area. The asbestos was identified in 7 rock and 3 soil samples collected in the serpentinite area. The geometric means of airborne concentrations of asbestos and nonasbestos fibers longer than 0.2 microm in length were: 384 and 447 fibers/liter (f/L) in the quarry property boundary (in operation), 12 and 124 f/L in the quarry property boundary (in closed), 5 and 103 fibers/liter in the serpentinite area, and less than 2 and 59 fibers/liter in the reference area, respectively. There was a significant difference in airborne concentrations of both asbestos and nonasbestos fibers among the areas. Ttremolite/actinolite were found with chrysotile in the stones. Airborne concentrations of tremolite/actinolite were higher than those of chrysotile at the quarry property boundary and in the serpentinite area. Tremolite/actinolite were also found in the soils. There was no significant difference among the areas in the arithmetic means of fiber size distribution of both asbestos and nonasbestos fibers.

An operated case of lung cancer with pleural plaques: its asbestos bodies, fiber analysis and asbestos exposure.

This case was a 79-year-old man with pleural plaques, which had been pointed out in the left lung field on chest X-ray six years ago. A new shadow in the right chest appeared in 1999 and was closely examined. Cytological class IV carcinoma was detected in his lung tissue obtained by broncho-fiberscope. Lobectomy of the right upper lobe was performed, and calcified pleural plaques were found on the chest wall. The clinical diagnosis was poorly differentiated squamous cell carcinoma, T1N0M0. In World War II when he was 26 years old, he had worked as a boiler man on a battle cruiser for one year. The amount of asbestos bodies (AB) was 3,348 per gram dry lung tissue. The cores of AB and asbestos fibers were examined and showed that amosite was the most prevalent and crocidolite, tremolite and chrysotile were present in that order. After leaving the navy, he had worked as a farmer throughout his life, suggesting that he had never contacted asbestos occupationally after being a boiler man. It is strongly suggested that he had been exposed to asbestos during his work as a boiler man and that produced pleural plaques and lung cancer 50 years' later.

Clinical, pathological and mineralogical features in two autopsy cases of workers exposed to agalmatolite dust.

An agalmatolite miner and processor showed large shadows at the bilateral hila accompanied by surrounding emphysematous changes and irregular shadows on chest X-ray films. Chest CT scans were characterized by a mixture of tiny irregular structures and small round opacities. Histopathological examination revealed massive fibrosis, which corresponded to large shadows, but only a small number of typical silicotic nodules, indicating mixed dust pneumoconiosis. Mineralogical examination of the autopsy lungs showed quartz, pyrophyllite, mica, and kaolinite. Quartz accounted for 70% of the amount of all mineral dust in both patients, but pyrophyllite accounted for 10.8% and 14.4%. The pulmonary mineral dust composition in the two patients was well consistent with the mineral composition of the raw clays in the agalmatolite mine. In the two patients, chest X-ray findings and histopathological findings of the lungs also suggested agalmatolite pneumoconiosis, which was confirmed by mineral analysis of the lungs.

Biological effects of man-made mineral fibers (II)--their genetic damages examined by in vitro assay.

In order to study and compare genetic damage induced by 10 kinds of man-made mineral fibers (JFM fibers) in cells, human lung epithelial cells (A549) were exposed to JFM fibers and chrysotile for 1 h, then single-cell gel electrophoresis (SCGE) assay was used to detect DNA strand breaks, DNA-DNA interstrand crosslink and the ability of DNA to repair; The results showed that all 10 JFM fibers could induce DNA strand breaks, DNA-DNA interstrand crosslinks and inhibit the ability of DNA repair. When human embryo lung (HEL) cells were exposed to JFM fibers and chrysotile for 24 h respectively, the chromosomal aberration was analyzed and the results showed that chrysotile and most of JFM fibers at 5.0 micrograms/ml induced structural chromosomal aberration, but all of these effects were lower than that of chrysotile and were different among them, suggesting that 10 types of JFM fibers had genotoxicity with different degree in vitro, but all of them were lower than that of chrysotile.

Acute biological effects of intratracheally instilled titanium dioxide whiskers compared with nonfibrous titanium dioxide and amosite in rats.

The dimensions of man-made mineral fiber whiskers are similar to those of some kinds of asbestos. Thus these mineral fibers raise the concern for potential health hazard for workers exposed in the occupational environments. This study was designed to define acute biological effects of intratracheally administered titanium dioxide whiskers (TO1) compared with nonfibrous titanium dioxide (TOP) and UICC amosite (Ams), and their relations to acute lung inflammation in rats. The observed geometric mean length (microm) and width (microm) and geometric standard deviation are: TO1(2.1[2.0], 0.14[1. 53]); Ams (4.3[3.3], 0.31[1.9]); and TOP (50 nm, 1-2 microm aggregates). Ten-week-old Wistar-Jcl male rats received a single tracheal injection of test materials at doses between 0.05 and 1.0 mg/rat. Control animals were injected with the same volume of saline. Lung tissue and bronchoalveolar lavage (BAL) fluid were collected from rats on days 1, 3, and 7 after administration. In the group injected with TO1, total protein, cytokine-induced neutrophil chemoattractant (CINC)/growth-regulated gene product (GRO), interleukin (IL) 1beta, and tumor necrosis factor (TNF) alpha increased on day 1. Subsequently, total elastolytic activity and fucose levels in BAL increased by day 3. All parameters, except for fucose in BAL, recovered to the normal levels. Animals in the Ams group showed increased total protein and CINC/GRO and decreased total elastolytic activity in a dose-dependent manner on day 1. The fucose level increased on day 3 in the Ams group. All parameters returned to their control levels on day 7. Animals in the TOP group did not show significant changes any of parameters during the experimental period. Gene expression of TNF-alpha and monocyte chemoattractant protein (MCP) 3 in the lung increased dose-dependently in the animals treated with the three materials. The mRNAs for eotaxin and MIP-1alpha were overexpressed in the lung of animals treated with Ams and TO1, while RANTES mRNA was overexpressed dose-dependently in the lung of animals treated with Ams on day 1. Onset of inflammatory response was more rapid in the Ams group than the TO1 group. Recovery of the fucose level in BAL was slower in the TO1 group than in the Ams group, though we observed similar histopathological changes in the lung of animals with TO1 or Ams. We conclude that whisker-induced acute biological effects in the lung may be related to the shape of the whiskers and not to their chemical composition or surface crystal structure, showing biological effects similar to those of UICC amosite.

A retrospective cohort study of male workers exposed to PVA fibers.

In order to ascertain whether PVA fibers can produce cancer in humans or not, we have conducted a retrospective cohort study of workers exposed to PVA fibers. A total of 447 exposed and 2416 non-exposed male workers who were engaged before 1980 were followed up until the end of 1996. The SMR for all causes was 0.57 (observed 38, 95% CI: 0.41-0.78) for the exposed, and 0.66 (observed 210, 95% CI: 0.58-0.75). As for lung cancer, its SMR was 0.77 (observed 3, 95% CI: 0.15-2.24) for the exposed workers and 0.67 (12 observed, 95% CI: 0.34-1.16) for the non-exposed workers. Lung cancer SMR was 0.86 (observed 2, 95% CI: 0.10-3.11) for the workers with 20 or more years' employment. This study showed no difference in lung cancer risk between the workers exposed to PVA fibers and the non-exposed workers.

Hexavalent chromium responsible for lung lesions induced by intratracheal instillation of chromium fumes in rats.

Lung toxicity of chromium fumes (Cr fumes) was examined by a single intratracheal instillation into rats of 10.6 mg and 21.3 mg Cr fumes/kg body weight and by repeated (3 times) instillations of 10.8 mg and 21.7 mg Cr fumes/kg. The pathological changes were compared with those induced by single administrations of 3.2 mg and 19.2 mg Na2CO3 solution-insoluble fraction of Cr fumes (Cr-Fr)/kg and 20.8 mg commercially available chromium (III) oxide powder (Cr (III) oxide)/kg. Single and repeated administrations of Cr fumes suppressed growth rate in a dose-dependent manner, but administrations of Cr-Fr and Cr (III) oxide did not. A single administration of Cr fumes produced granulomas in the entire airways and alveoli with progressive fibrotic changes, as well as severe mobilization and destruction of macrophages and foamy cells. Those histopathological changes were aggravated by the repeated administration of Cr fumes. On the other hand, single administrations of Cr-Fr and Cr (III) oxide produced no remarkable histopathological changes. Cr fumes were found to be composed of 73.5% chromium (III) oxide and 26.5% chromium (VI) oxide. The primary particles of Cr fumes and Cr-Fr were similar, 0.02 micron in size (sigma g: 1.25), and Cr (III) oxide particles were 0.30 micron in size (sigma g: 1.53), measured by analytical electron microscopy (ATEM). Diffuse clusters of the primary particles in Cr fumes were identified as Cr (VI) oxide. The present results suggested that the lung toxicity of Cr fumes was mainly caused by these Cr (VI) oxide (CrO3) particles in Cr fumes.

Biological effects of man-made mineral fibers (I)--Reactive oxygen species production and calcium homeostasis in alveolar macrophages.

10 types of standard mineral fiber samples (JFM fibers) were tested for their cytotoxicity in alveolar macrophages (AM) in vitro experiments, in which UICC chrysotile B was used as a positive control. The cytotoxicity included the production of superoxide anion radical and hydrogen peroxide, depletion of glutathione (GSH) and increase of intracellular free calcium. The results showed that chrysotile and most of the 10 mineral fibers could increase the production of superoxide anion and hydrogen peroxide, deplete the concentration of GSH and increase the level of free intracellular Ca2+ in AM. But all the effects of JFM fibers were lower than that induced by UICC chrysotile B. Although the cytotoxicity of JFM fibers were lower than that of asbestos, these mineral fibers should be used with highly care for workers in industries.

Anthophyllite exposure and endemic pleural plaques in Kumamoto, Japan.

OBJECTIVES: This study explored the high prevalence of pleural plaques in the town of Matsubase in Kumamoto, Japan. METHODS: Small-size chest X-ray film was used for screening, and all persons with pleural plaques were confirmed by computed tomography (CT). The prevalence rate of pleural plaques in the 4 districts of Matsubase and its surrounding towns and cities were also examined. The age-adjusted mortality rate for lung cancer in this town was compared with that of its surrounding towns and cities. RESULTS: Pleural plaques were found in 1357 persons (724 men and 633 women) among the inhabitants who were more than 20 years of age in Matsubase between 1988 and 1993. CT scans ascertained 938 cases with pleural plaques among the 11 14 persons who participated. Thus at least 9.5% of the inhabitants over 20 years of age in this town had pleural plaques. The neighboring towns had a higher rate than the more distant towns. A large-scale open-cast asbestos mine and mill had been in operation in Matsubase between 1883 and 1970. Mineral analysis revealed anthophyllite fibers. Most of the plaques were found in persons who had never worked in the mine or mill. CONCLUSIONS: The high prevalence of pleural plaques in Matsubase was due to anthophyllite exposure, mainly environmental. No mesotheliomas were found, however. These findings agree with those from an earlier study from Finland.

Fiber numbers per unit weight of JFM standard reference samples determined with a scanning electron microscope. Japan Fibrous Material.

A standard reference 10 sample-set of fibrous minerals were prepared by the Japan Fibrous Material Research Association (JFMRA) under the name of JMF standard reference samples. In this paper, the fiber number per unit weight of JFM standard reference samples was determined with a scanning electron microscope. Fiber numbers per unit weight (f/microgram) and standard deviations observed in this experiment were as follows: glass wool (GW1) 7.0 +/- 0.1 x 10(2), rock wool (RW1) 1.7 +/- 0.2 x 10(3), micro glass fiber (MG1) 6.5 +/- 0.4 x 10(4), refractory ceramic fibers; (RF1) 8.8 +/- 0.7 x 10(3), (RF2) 8.7 +/- 0.8 x 10(3), mullite fibers (RF3) 3.5 +/- 0.7 x 10(3), potassium titanate whisker (PT1) 5.9 +/- 0.3 x 10(5), silicon carbide whisker (SC1) 4.1 +/- 0.4 x 10(5), titanium oxide whisker (rutile) (TO1) 6.4 +/- 0.6 x 10(5), and wollastonite (WO1) 2.4 +/- 0.1 x 10(4). Fiber numbers per unit weight would change in proportion to the cube or cube root of the fiber size if the fibers have the same density and the same aspect ratio. JFM standard reference samples should be used taking into consideration the difference in fiber number per unit weight when users conduct in vitro and/or in vivo (injection) biological experiments using these samples.

Differences in the extent of inflammation caused by intratracheal exposure to three ultrafine metals: role of free radicals.

Nickel and cobalt, which belong to the same elemental group, are known to cause interstitial lung disease and bronchial asthma. The ability of these metals to injure lung cells and cause inflammation is likely to be important in their pathogenicity but comparative studies are rare. Additionally, ultrafine (uf) forms of these metals are used increasingly and there is little available information on their toxicity. Thus the inflammatory response following intratracheal instillation of ultrafine particles of Co, Ni, and TiO2 was compared. Physiological saline (PS) was used as a vehicle control and DQ12 quartz as a positive control. Male Wistar rats were intratracheally instilled with the 3 particle types at a dose of 1 mg suspended in physiological saline. At 1, 3, 7, 15, and 30 d after the injection, lung weight and the cellular and biochemical changes in bronchoalveolar lavage fluid (BALF) were determined. By all of the indices, Uf-Ni appeared to be the most injurious to the lung, causing severe and sustained inflammation, cytotoxicity and increased epithelial permeability. The next most toxic material was DQ12 quartz, with Uf-Co being closely similar in ability to cause inflammation. Uf-TiO2 was more active than the saline control in all of the indices, but was the least toxic of the particles studied. The present study reveals that three ultrafine particles of the same diameter are dramatically different in their ability to cause inflammation. The three ultrafines were compared as to their ability to cause free-radical damage to supercoiled plasmid DNA, and the result of free-radical activity was found to be Uf-TiO2 << Uf-Co = Uf-Ni. Difference in free-radical-generation activity therefore could underlie the difference in inflammation of these three ultrafine particle types.

Preparation and characteristics of standard reference samples of fibrous minerals for biological experiments.

Standard reference samples of fibrous minerals to be used for in vitro and in vivo (injection) biological experiments were prepared. Ten samples were selected from among the fibrous minerals currently used in industry: man-made mineral fibers (MMMF); glass wool (GW1), rock wool (RW1), micro glass fiber (MG1), refractory ceramic fibers (RF1, RF2), and mullite fibers (RF3), whiskers; potassium titanate (PT1), silicon carbide (SC1) and rutile (TlO2)(TO1), and a natural fibrous mineral; wollastonite (WO1). The MMMF samples were prepared by using the size-selective press method. The whisker samples and WO1 were all commercial products. The prepared samples were examined by X-ray diffraction and X-ray fluorescence analyses, and no impurities were identified in any of the samples at a level of 1%, although WO1 was found to have about 5% calcite and a trace amount of quartz (< 1%). The fiber sizes were measured by TEM. The geometric-mean length (micron), width (micron) and SD (in parenthesis) are: GW1 20.0 (2.58), 0.88 (3.10); RW1 16.5 (2.51), 1.80 (2.32); MG1 3.0 (2.22), 0.24 (2.35); RF1 12.0 (2.36), 0.77 (2.53); RF2 11.0 (1.96), 1.10 (2.00); RF3 11.0 (1.75), 2.40 (1.37); PT1 6.0 (2.04), 0.35 (1.51); SC1 6.40 (2.45), 0.30 (1.58); TO1 2.1 (2.00), 0.14 (1.53); WO1 10.5 (2.03), 1.00 (1.72). We concluded that these prepared fiber samples can be safely used for biological experiments. Sample sets containing about one gram of each sample known as JFM standard reference samples are provided by the Japan Fibrous Material Research Association (JFMRA).

Inhibition of arachidonate lipoxygenase activities by 2-(3,4-dihydroxyphenyl)ethanol, a phenolic compound from olives.

The effects of olive fruit extract on arachidonic acid lipoxygenase activities were investigated using rat platelets and rat polymorphonuclear leukocytes (PMNL). Olive extract strongly inhibited both 12-lipoxygenase (12-LO) and 5-lipoxygenase (5-LO) activities. One of the compounds responsible for this inhibition was purified and identified as 2-(3,4-dihydroxyphenyl)ethanol (DPE). DPE inhibited platelet 12-LO activity (IC50, 4.2 microM) and PMNL 5-LO activity (IC50, 13 microM) but not cyclooxygenase activity in cell-free conditions. It also inhibited 12-LO activity in intact platelets (IC50, 50 microM) and reduced leukotriene B4 production in intact PMNL stimulated by A23187 (IC50, 26 microM). The inhibition by DPE of both lipoxygenase activities was stronger than that by oleuropein, caffeic acid, or 7 other related phenolic compounds, especially in intact cells. These results suggest that DPE is a potent specific inhibitor of lipoxygenase activities.

Length-reduction method for man-made mineral fibers for biological experiments.

A simple fiber length-reduction method was developed to obtain a large amount of fiber samples with different length distributions for use in various biological experiments. This press method is only to press a raw fiber sample charged in a stainless cylinder at an adequate pressure, and is effective for man-made mineral fibers (MMMF) such as glass wool, rock wool and wool and refractory fibers (ceramic fibers, mullite fibers) and some brittle natural mineral fibers such as fibrous brucite and wollastonite. The mean fiber-length of man-made mineral fibers became shorter with the increase in the pressure applied without diameter change. We could obtain a length-reduced fiber sample with a suitable length distribution by this method. This press method is therefore a size-selective method able to produce a large amount of pulverized fiber sample depending on the press cylinder size for biological experiments. A very small amount of non-fibrous particles with aspect ratios (length vs. diameter) of under 3 was seen in the pulverized fiber samples. To eliminate such non-fibrous particles as well as too long fibers from the sample, separation by sedimentation in water was somewhat effective.

Mineral phases and some reexamined characteristics of the International Union Against Cancer standard asbestos samples.

Standard asbestos samples to be used for biomedical research were first prepared by the International Union Against Cancer (UICC) in 1966 in the United Kingdom and South Africa. Using modern techniques, X-ray diffractometry, analytical transmission electron microscopy, and thermal analysis, we have now analyzed these UICC samples to determine the mineral compositions (mineral phases) and their respective quantities. UICC chrysotile A (from Zimbabwe) contains 2% fibrous anthophyllite as impurity; chrysotile B (from Canada) does not contain any fibrous impurities, only non-fibrous minerals. UICC amosite and crocidolite are almost pure. UICC anthophyllite has 20-30% talc as impurity. The chemical compositions and fiber size distributions of the UICC asbestos samples have also been determined. The mean widths of the fibers of chrysotile A and B are smaller than those of the amphibole fibers. This agrees well with the earlier results which showed the two chrysotile samples to have a larger respirable fraction than the amphiboles.

[Concentration and fiber size of asbestos in lungs of residents living close to the serpentinite area].

Concentrations and fiber size distribution of asbestos in the lung tissue of 16 ( 8 males and 8 females) rural residents living close to the serpentinite area (neighboring group) were analyzed by low temperature ashing-analytical transmission electron microscopy, and were compared with those of 5 (5 males) rural and 53 (34 males and 19 females) urban residents living away from the serpentinite area (reference group). The results are as follows; 1. Pulmonary asbestos fibers were found in all rural residents. The types of asbestos fibers were chrysotile, amosite, crocidolite, actinolite, and tremolite. These findings were very similar to results for urban residents. 2. In males, the geometric mean concentration of actinolite-tremolite in the neighboring group (1.78 x 10(6) fibers/g dry lung) was significantly higher than those in the reference groups (rural: 0.32 x 10(6), urban: 0.31 x 10(6), p < 0.01). In females also, the geometric mean concentration of actinolite-tremolite in the neighboring group (0.59 x 10(6)) was significantly higher than that in the reference group (urban: 0.16 x 10(6), p < 0.05). There were no significant differences in the actinolite-tremolite concentrations between males and females in the neighboring group. 3. The serpentinite in this area have contained not only chrysotile but also both actinolite and tremolite. There was no significant relationship between actinolite-tremolite concentration and occupational history in the neighboring group. It is possible that actinolite and tremolite are liberated from the serpentinite to the atmosphere and inhaled by the residents near the serpentinite area which increases pulmonary actinolite-tremolite concentration. 4. The difference in the geometric mean concentration of chrysotile between the neighboring and the reference group was not statistically significant for both sexes, possibly due to the poor durability of chrysotile in the lung tissue. 5. There were no significant differences in the geometric mean of fiber length and diameter of actinolite-tremolite between the neighboring and the reference group.