The Exploitation and Environmental Legacy of Amphibole Asbestos : A Late 20th Century Overview

The risk to human health associated with the inhalation of amphibole asbestos has become devastatingly ap- parent this century. The most commonly utilised asbestiform amphiboles, crocidolite (blue asbestos) and amosite (brown asbestos), are implicated in a variety of diseases related to cell damage within the respiratory tract and adjacent areas. Blue and brown asbestos fibres have a morphology and mineralogy which makes them more biopersistent and biochemically reactive than chrysotile (white asbestos). The long-term pres- ence of such fibres within and around the lungs can result in fibrous scarring, lung cancer, and is the major cause of the once extremely rare tumour known as diffuse malignant mesothelioma. Therefore, despite the fact that asbestiformamphiboles comprise approximately 5%of industrially utilised asbestos (the rest being chrysotile), they have been disproportionately pathogenic. Almost all amphibole asbestos sold on the world market was mined from Palaeoproterozoic ironstones in either the northern Cape Province (blue) or Trans- vaal (brown and blue) areas in South Africa. Production peaked from 1966–1978 when around 2 million tons of crocidolite and1million tons of amosite were produced and exported to be used mainly in asbestos-based cement products and many types of building materials. Crocidolite mixed with chrysotile was commonly used in pressure pipes and gaskets, whereas amosite mixed with chrysotile was especially suitable for gut- ters, roofing, and insulation boards. Amid- to late 20th century amphibole asbestos-related cancer epidemic has consequently struck not only mining and milling communities in producer countries, but many groups of workers (and their relatives and neighbours) exposed to amphibole asbestos-bearing materials in importer countries. Although belated closure and reparation of the mines and imposition of threshold safety limits in the workplace will eventually stem this epidemic the death toll has not yet peaked. Given the long latency period (decades) typical of mesothelioma and bronchogenic carcinoma and the fact that amphibole asbes- tos-bearing materials are still present in some buildings, asbestos-related cancer will inevitably continue to be a major cause of death in many countries worldwide well into the next century.     

Cytotoxicity of single-walled carbon nanotubes,multi-walled carbon nanotubes,and chrysotile to human lung epithelial cells

Carbon nanotubes (CNTs) have found numerous applications in various industries. Recently, adverse effects of these materials on human and animal cells in vitro have been reported. In the present study, the cytotoxicity of single-walled carbon nanotubes (SWCNTs), multi-walled carbon nanotubes (MWCNTs), and chrysotile asbestos in human lung epithelial cells has been studied using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. The cells were exposed for 6 h and 24 h to between 0.97 and 1500 μg mL^?1 of CNTs and chrysotile fibers prepared in two culture media containing 5% serum and 0.5% dimethylsulfoxide. Dose–response curves were obtained to determine the nonobservable adverse effect concentration and the half-maximum inhibitory concentration (IC₅₀). The way of dispersion affects the cytotoxicity of CNTs. For MWCNT, the toxicological indexes were lower than for SWCNT. Chrysotile fibers were even less cytotoxic than CNTs. Therefore, workplace control measures are recommended as priority for occupational and environmental conditions.     

A state of the science review of the potential health hazards associated with asbestos in shielded metal arc welding rods in the United States

Due to its unique chemical properties, chrysotile asbestos was historically incorporated into a wide variety of products, including the outer covering, or “flux” of certain classifications of general arc mild steel welding electrodes. The purpose of this analysis is to review the historical engineering, toxicology, regulatory, and epidemiology information relevant to asbestos in mild steel welding rods in order to assess whether mild steel welders are at increased risk of developing asbestos-related diseases as a result of welding rod use. We divided our analysis into four distinct time periods, based on what we perceived to be seminal events in welding technology or in the evolution of knowledge regarding asbestos and asbestos-related diseases. These time periods are as follows: late 1800s to 1929, 1930–1955, 1956–1970, and 1971–2006. We found that studies that attempted to measure airborne asbestos directly in welding rod fumes found no measurable fibers; this is likely due to the fact that the fibers degrade at high temperatures present in the welding arc. In addition, “worst-case” use simulation studies, specifically intended to generate airborne flux particles, reported that airborne asbestos concentrations were either undetectable or very low. The airborne concentrations generated were always below the current OSHA permissible exposure limit (0.1 f/cc TWA), and the lifetime doses associated with “worst-case” use were found to be far below the plausible thresholds for mesothelioma and lung cancer. The epidemiology of mild steel welders is difficult to interpret due to confounding exposures such as (1) possible exposure to nickel and hexavalent chromium in stainless steel welding fumes (lung cancer), (2) smoking (lung cancer), and (3) bystander exposure to amphibole asbestos (lung cancer and mesothelioma). None of the published welder studies controlled for all three factors, yet when any single source of bias was controlled for, the majority of the studies reported no significant risk in either asbestos-related disease. Furthermore, none of the investigators in any of the welder studies suggested that asbestos in welding rods might be a risk factor for lung cancer or mesothelioma, and there is not a single case report in the medical literature that attempts to link welding rod use to an asbestos-related disease. We conclude that the weight of evidence indicates that welders were not historically at risk of developing asbestos-related diseases as a result of welding rod use.     

State-of-the-science assessment of non-asbestos amphibole exposure: Is there a cancer risk?

The distinction between amphibole asbestos fibers and non-asbestos amphibole particles has important implications for assessing potential cancer risks associated with exposure to amphibole asbestos or amphibole-containing products. Exposure to amphibole asbestos fibers can pose a cancer risk due to its ability to reside for long periods of time in the deep lung (i.e., biopersistence). In contrast, non-asbestos amphibole particles are usually cleared rapidly from the lung and do not pose similar respiratory risks even at high doses. Most regulatory and public health agencies, as well as scientific bodies, agree that non-asbestos amphiboles possess reduced biological (e.g., carcinogenic) activity. Although non-asbestos amphibole minerals have been excluded historically from Federal regulations, non-asbestos structures may be counted as asbestos fibers on the basis of dimensional criteria specified in analytical protocols. Given the potential to mischaracterize a non-asbestos structure as a “true” asbestos fiber, our objective was to assess whether exposure to non-asbestos amphiboles that may meet the dimensional criteria for counting as a fiber pose a cancer risk similar to amphibole asbestos. We reviewed analytical methods as well as the mineralogical, epidemiological, and toxicological literature for non-asbestos amphiboles. No evidence of demonstrable cancer effects from exposure to non-asbestos amphiboles that may be counted as fibers, under certain assessment protocols, was found. Data gaps (industrial hygiene data for amphibole-exposed cohorts), inconsistencies (analytical laboratory methods/protocols used to count fibers), and sources of potential bias from misclassification of exposure were identified.     

Characterization and in vitro biological effects of ambient air PM10 from a rural,an industrial and an urban site in Sulaimani City,Iraq

Abstract

High urban atmospheric pollution is caused by economic and industrial growth, especially in developing countries. The objective of this study was to assess possible relationships between in vitro effects on human alveolar epithelial cells of source-related dust types collected at Sulaimani City (Iraq), and to determine their mineralogical and chemical composition. A passive sampler was used to collect dust particles at a rural, an industrial and an urban sampling site during July and August 2014. The samples were size-fractionated by a low-pressure impactor to obtain respirable dust with aerodynamic diameters of less than 10?µm. The dust was mainly composed of quartz and calcite. Chrysotile fibers (white asbestos) were also found at the urban site. Dust from the industrial and urban sites triggered cytotoxic and genotoxic effects in the cells, whereas only minor effects were observed for the sample from the rural site.     

Erionite series minerals: mineralogical and carcinogenic properties

Erionite is a human and animal carcinogen and one of the most toxic minerals known. Erionite deposits have been reported in many countries; however, it is only in the area of three villages of Cappadocia, Turkey, that environmental exposure to erionite has been demonstrated to be the cause of an epidemic of the disease mesothelioma. In the USA, no cases of mesothelioma have been reliably proven to be the result of erionite exposure, though the possibility exists. Erionite samples from three villages of the Cappadocia region were characterized mineralogically and compared with three different standards from the USA. Micro morphological details of erionite minerals using a high-resolution field-emission SEM showed that microstructures of "bundles", "fibers", and "fibrils" are important physical properties of fibrous erionite minerals. Typical lung burden of erionite and asbestos fibers were compared in terms of number of fibers. Assuming the lung burden of fibers in a human mesothelioma victim is about 1 mg, and the hazardous fibers are approximately 1 mum in diameter and 10 mum long, that milligram contains approximately 40 million asbestos and 50 million erionite fibers. These microstructures of erionite minerals draw attention to the concepts of surface area or surface-area-to-volume ratio and their relationship to the carcinogenicity of the mineral. The larger surface area creates a wider platform for mineral-cell interaction and thus more possibilities of proliferative transformation of mesothelial cells. Consequently, understanding the exact mineralogical properties will help determination of the true carcinogenic mechanism(s) of the mineral for prevention and possibly treatment of malignant mesothelioma.     

Lipid peroxidation in the presence of iron oxides

Exposure to iron oxides dusts may lead to lung cancers among exposed population. To evaluate the involvement of iron‐containing particles in lipid peroxidation by production of reactive oxygen species, hematite, magnetite (iron oxides), and crocidolite (asbestos compound) were tested. The peroxidation was followed by the evaluation of some degradation products of lino‐lenic acid. In a buffered medium, magnetite is higher active than hematite. The addition of an iron chelator (EDTA) or of a reducing agent (ascorbate) in this medium enhances the activity of hematite and magnetite, and the combination of both EDTA and ascorbate increases their activity. Crocidolite is the most active whatever the medium. The appearance of the EDTA‐iron‐oxygen complexes related to the reactivity of these oxides is postulated. These results suggest that the oxidizing activity triggered by hematite and magnetite, in a medium containing an iron chelator and a reducing agent, may lead to damages in biological medium.     

The assessment of the malignant mesothelioma cases and environmental asbestos exposure in Sivas province, Turkey

One of the most significant diseases related to environmental asbestos exposure is malignant mesothelioma (MM). Sivas province is located in the Central Anatolia where asbestos exposure is common. We aimed to study clinical, demographical and epidemiologic features of the patients with MM in Sivas, along with the history of asbestos exposure. In total, 219 patients with MM who were diagnosed in our hospital between 1993 and 2010 were retrospectively analyzed in terms of demographical and clinical features. Rock, soil and house plaster samples were taken from the habitats of those patients and were evaluated with optical microscopy and X-ray diffraction methods. The age of the patients ranged between 18 and 85 years. The male-to-female ratio was 1.4:1. Most of the patients confirmed an asbestos exposure history. The most frequent symptoms of the patients were chest pain (60 %) and dyspnea (50 %). The gap between the start of first symptoms and the diagnosis date was approximately 4 months in average. The plaster materials used in most of the houses were made up of mainly carbonate and silicate minerals and some chrysotile. Ophiolitic units contained fibrous minerals such as serpentine (clino + orthochrysotile) chiefly and pectolite, brucite, hydrotalcite and tremolite/actinolite in smaller amounts. MM is not primarily related to occupational asbestos exposure in our region, and hence, environmental asbestos exposure may be indicted. Yet, single or combined roles and/or interactions of other fibrous and non-fibrous minerals in the etiology of MM are not yet fully understood and remain to be investigated.     

论城市室内环境中气溶胶污染问题

本文全面评述了室内环境中气溶胶的种类来源，化学组成，形成和自然清除机理。空气环境中气溶胶大致可分为：１．燃烧型气溶胶，主要包括各类燃料燃烧产生的烟雾和烟草烟雾等；２．矿物型气溶胶，包括粉尘，飞灰，石棉及其它天然纤维尘等；３．生物型气溶胶，包括植物生气溶胶，如动物皮屑及各类微生物等。     

纳米二氧化钛暴露人胚肺细胞差异表达基因相关通路

为探讨纳米二氧化钛(Nano-TiO2)对人胚肺(HPF)细胞差异表达基因相关通路的影响,采用半致死浓度(0.437mg·mL-1)的10nmTiO2暴露体外培养的人胚肺细胞24h,提取RNA,应用基因芯片技术筛选差异表达基因,分析纳米TiO2对基因通路的影响.结果表明,纳米TiO2暴露人胚肺细胞,导致514条肺中表达的基因发生差异表达,涉及多个KEGG通路和BioCarta通路.纳米TiO2暴露人胚肺细胞可能产生以下生物学效应:1)众多位于细胞外区域和细胞膜上的基因(特别是细胞膜受体基因)差异表达,对外界环境胁迫产生应激反应;2)与炎症相关的细胞因子基因表达大量改变,调控细胞的炎症反应;3)钙离子通路的膜受体基因差异表达,导致大量钙离子内流,调控钙离子信号传导;4)某些基因的差异表达(如OCLN下调),降低了细胞紧密联接力;5)与造血细胞增殖和分化相关的基因差异表达,刺激产生大量白细胞以抵抗感染.     

纳米二氧化钛暴露人胚肺细胞差异表达基因分析

为探讨纳米二氧化钛(Nano-TiO2)颗粒对人胚肺(HPF)细胞基因表达和基因功能的影响,使用粒径10nmTiO2暴露体外培养的人胚肺细胞24h,提取RNA,应用基因芯片方法,寻找差异表达基因,并对差异基因进行基因本体(GeneOntology,GO)分类.结果表明,纳米TiO2暴露人胚肺细胞,导致514条肺中表达的基因发生差异表达,基因分类显示400条基因涉及生物学过程;415条基因涉及分子学功能;391条基因涉及细胞构成.纳米TiO2作为外界刺激物质,与细胞膜上的受体结合,影响钙、钾离子通道,上调细胞免疫和炎症反应相关的细胞因子TNF、IL1B、IL1A等基因表达.     

甲醛对大鼠肺细胞醛糖还原酶的影响及醛糖还原酶功能研究

为了研究甲醛作用后大鼠肺细胞醛糖还原酶(AR)的活性变化以及AR在肺细胞损伤中的功能,首先通过大鼠肺细胞分离培养,观察甲醛对大鼠肺细胞AR活性的影响及AR抑制剂——盐酸小檗碱对AR活性的抑制作用;其次通过流式细胞仪检测甲醛及盐酸小檗碱对大鼠肺细胞周期和凋亡的影响.结果表明:1.0mmol·L-1甲醛作用24h使大鼠肺细胞AR活性显著增加,盐酸小檗碱对AR活性具有抑制作用;甲醛能够引起大鼠肺细胞凋亡,使G0/G1和S期的细胞比例下降,G2/M期的细胞比例增加;甲醛和盐酸小檗碱共同作用使大鼠肺细胞凋亡率较单纯甲醛组更高,同时G0/G1期的细胞比例增加,S和G2/M期的细胞比例下降.结果提示甲醛能够使大鼠肺细胞AR活性增加;AR活性增加可减少细胞凋亡,对甲醛引起的大鼠肺细胞损伤具有保护作用.     

Effects of three different nanoparticles on bioaccumulation,oxidative stress,osmoregulatory, and immune responses of Carcinus aestuarii

Abstract

In this study, the toxicity of CuO (40?nm), α-Al₂O₃ (40?nm), and α-Fe₂O₃ (20–40?nm) nanoparticles was comparatively investigated on Carcinus aestuarii. Crabs were semi-statically exposed to 1?mg/L of each for 14?days and their accumulation and distribution in tissue and hemolymph, potential oxidative stress mechanism, total hemocyte counts and types, and the osmoregulatory and ionoregulatory responses were determined. The tissue distribution of CuO nanoparticles was hepatopancreas?>?hemolymph?≥?gill?> muscle, for α-Fe₂O₃ gill?>?hepatopancreas?>?muscle?> hemolymph, and for α-Al₂O₃ gill?>?muscle?≥?hemolymph?> hepatopancreas. While α-Al₂O₃ and α-Fe₂O₃ NPs, induced lipid peroxidation and changes in antioxidant enzyme activity in the hepatopancreas tissue, the oxidative damage caused by the CuO nanoparticles was minimal. All three nanoparticles, copper in particular, elicit osmoregulatory and ionoregulatory toxicity at this concentration, due to the inhibition of Na⁺, K⁺-ATPase activity in the gill and depletion of hemolymph and carcass ion concentrations.     

Raw single-walled carbon nanotube-induced cytotoxic effects in human bronchial epithelial cells: comparison to asbestos

Single-walled carbon nanotubes (SWCNT) are being developed to be used in many industrial and biomedical applications. However, SWCNT's durability and likely fibrous morphology have raised health concerns. The present investigations were focused on understanding the cellular and molecular mechanisms induced by raw SWCNT (SWCNT) in human bronchial-epithelial cells (BEAS-2B). Asbestos (crocidolite) was used as a positive control. Exposure of BEAS-2B cells to SWCNT induced apoptosis, DNA damage, and oxidative stress. The generation of hydroxyl radical (^?OH) and increase of superoxide dismutase (SOD) activity were concentration-dependent. The increase in apoptosis was associated with activation of caspase-3, caspase-7, and poly (ADP-ribose) polymerase-1 (PARP-1). A short recovery period of 6?h of cells from SWCNT exposure resulted in reversal of caspase-3 and caspase-7, and a partial reversal of PARP-1 activation. The activation of PARP-1, caspase-3, and caspase-7 was only partially diminished after a recovery of 6?h from the exposure to crocidolite. Exposure of BEAS-2B cells to SWCNT resulted in the phosphorylation of protein p42/44 (p42/44) and protein p38 (p38). SWCNT did not induce protein serine-threonine kinase (AKT) phosphorylation. For all the above end points, crocidolite induced a greater response compared to SWCNT. SWCNT induced a significant activation of activator protein-1 (AP-1) and nuclear factor kappa B (NF-κB), and the effect was inhibited by mitogen-activated protein kinase (MAPK) inhibitors. SWCNT also induced significant increase in the expression levels of c-Jun, βIGH3, and CD44 genes. The results of this study show that the molecular mechanism for raw SWCNT-mediated toxicity in BEAS-2B cells is through the activation of caspase-3, caspase-7, and PARP-1. Furthermore, the mechanism of AP-1 and NF-κB activation is through MAPK. This bioactivity of raw SWCNT is associated with the generation of oxidative stress and DNA damage. Considering the role of airway epithelium as a critical barrier for normal pulmonary function and focal point for tumor development, this study demonstrates that raw SWCNT activate molecular events which may be linked to adverse biological responses implicated in pulmonary diseases.     

诺氟沙星对海月水母螅状体Hsp70基因、GTP结合蛋白基因和氧化应激蛋白基因表达的影响

诺氟沙星(norfloxacin,NFLX)广泛应用于水产养殖中的鱼类细菌性疾病治疗。为探讨诺氟沙星对海洋生物的毒性作用,选择海月水母螅状体为受试生物,考察了不同浓度诺氟沙星和不同暴露时间对GTP结合蛋白(GTP binding protein)、氧化应激蛋白(oxidative stress protein)和热休克70 k Da蛋白(heat shock 70 k Da protein,Hsp70)表达量的影响。结果表明,NFLX对海月水母螅状体的48 h的半致死浓度(48 h-LC_(50))是415.1 mg·L~(-1),NFLX对海月水母螅状体的毒性属于低毒。随着NFLX浓度的升高和培养时间的延长,Hsp70基因在第7天浓度300 mg·L~(-1)时表达量受到显著性诱导(P0.05),较对照组升高9.1倍;GTP结合蛋白基因和氧化应激蛋白基因的表达量都呈现先急剧升高后降低的趋势。2个基因均在第1天受到显著性诱导(P0.05),分别在NFLX浓度100 mg·L~(-1)和300 mg·L~(-1)时表达量达到最大值,较对照组升高10.15倍和50.5倍。Hsp70基因、GTP结合蛋白基因和氧化应激蛋白基因在实验期间都对NFLX表现出较好的响应。     

Effects of perfluorooctanoic acid on neural genes expression and neuronal morphology in the planarian Dugesia japonica

Perfluorooctanoic acid (PFOA) is an emerging persistent organic pollutant that is considered as a neurotoxicant in animal studies. However, these neurotoxicological studies did not evaluate potential neural deficits in aquatic animals associated from PFOA exposure. In this study, the effects of PFOA on neural genes expression and neuronal morphology in the planarian Dugesia japonica have been investigated. The results show that PFOA exposure results in neural-related genes expression alteration and neuronal morphology defects in planarians. These results suggest that the planarian D. japonica is a good bioindicator for the detection and evaluation of PFOA effects on freshwater invertebrates. The basal phylogenetic position of planarians makes them a good indicator species that can predict toxicity for higher taxa, providing information for the overall ecological toxicity of PFOA in aquatic environment.     

Biomarkers indicate mixture toxicities of fluorene and phenanthrene with endosulfan toward earthworm (<Emphasis Type="Italic">Eisenia fetida</Emphasis>)

α-Endosulfan and some polycyclic aromatic compounds (PAHs) are persistent in the environment and can reach crop products via contaminated agricultural soils. They may even be present as mixtures in the soil and induce mixture toxicity in soil organisms such as earthworms. In this study, the combined toxicities of PAHs with α-endosulfan were determined in Eisenia fetida adults using an artificial soil system. α-Endosulfan and five PAHs were tested for their acute toxicity toward E. fetida in artificial soils. Only α-endosulfan, fluorene, and phenanthrene showed acute toxicities, with LC₅₀ values of 9.7, 133.2, and 86.2 mg kg^?1, respectively. A mixture toxicity assay was conducted using α-endosulfan at LC₁₀ and fluorene or phenanthrene at LC₅₀ in the artificial soils. Upon exposure to the mixture of fluorene and α-endosulfan, earthworms were killed in increasing numbers owing to their synergistic effects, while no other mixture showed any additional toxicity toward the earthworms. Along with the acute toxicity results, the biochemical and molecular changes in the fluorene- and phenanthrene-treated earthworms with or without α-endosulfan treatment demonstrated that enhancement of glutathione S-transferase activity was dependent on the addition of PAH chemicals, and the HSP70 gene expression increased with the addition of α-endosulfan. Taken together, these findings contribute toward understanding the adverse effects of pollutants when present separately or in combination with other types of chemicals.     

Repeated-dose toxicity attributed to aluminum nanoparticles following 28-day oral administration,particularly on gene expression in mouse brain

Nanotechnology is a rapidly growing industry that has elicited much concern due to the lack of available toxicity data. Aluminum oxide nanoparticles (AlNP) were listed as a high-priority group in the Organization for Economic Co-operation and Development (OECD) Steering Group for Test Guidelines. In this study, AlNP 35 ± 18.8 nm in size were administered daily at doses of 15, 30, or 60 mg k^?1 for 28 days. A significant decrease in white blood cells (WBC), neutrophils, lymphocytes, and monocytes was observed in the group treated with 60 mg kg^?1 of AlNP, accompanied by a significant increase in platelets. The concentration of aluminum (Al) rose significantly in the thymus, lung, and brain of the group treated with 60 mg kg^?1 of AlNP. However, no significant changes in histopathology were observed. The expression for feeding behavior, energy expenditure, and neurodegeneration-related genes were up-regulated more than twofold by 60 mg kg^?1 AlNP. Consequently, data suggest that exposure to AlNP may result in adverse health effects, including but not limited to growth inhibition, immunosuppression, and neurodegeneration.     

Biological properties of mud extracts derived from various spa resorts

Spa resorts are known for thousands of years for their healing properties and have been empirically used for the treatment of many inflammatory conditions. Mud is one of the most often used natural materials for preventive, healing and cosmetic reasons and although it has been used since the antiquity, little light has been shed on its physical, chemical and biological properties. In this study we examined the effect of mud extracts on the expression of adhesion molecules (CAMs) by endothelial cells as well as their effects on monocyte adhesion to activated endothelial cells. Most of mud extracts inhibited the expression of VCAM-1 by endothelial cells and reduced monocyte adhesion to activated endothelial cells, indicating a potent anti-inflammatory activity. Furthermore, the mud extracts were tested for their antimicrobial activity; however, most of them appeared inactive against S. aureus and S. epidermidis. One of the mud extracts (showing the best stabilization features) increased significantly the expression of genes involved in cell protection, longevity and hydration of human keratinocytes, such as, collagen 6A1, forkhead box O3, sirtuin-1, superoxide dismutase 1 and aquaporin-3. The present study reveals that mud exerts important beneficial effects including anti-inflammatory and anti-aging activity as well as moisturizing effects, implicating important cosmeceutical applications.     

Ocean acidification reduces biomineralization-related gene expression in the sea urchin, Hemicentrotus pulcherrimus

Although recent studies have demonstrated that calcification in a wide range of marine organisms is profoundly affected by CO₂-induced ocean acidification, the mechanism of this phenomenon is still unclear. To clarify the effects of ocean acidification on the calcification process at the molecular level, we evaluated the expression of three biomineralization-related genes in the sea urchin Hemicentrotus pulcherrimus exposed under control, 1,000, and 2,000?ppm CO₂ from egg to pluteus larval stage. We found that the expression of the gene msp130, which is proposed to transport Ca²⁺ to the calcification site, is suppressed by increased CO₂ at pluteus larval stage. Meanwhile, expression of the spicule protein matrix genes SM30 and SM50 was apparently not affected. The results suggest that the combined effects of ocean acidification on the expression of skeletogenesis-related genes as well as the change in seawater carbonate chemistry affect the biomineralization ability of sea urchins.