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  • Hydrothermal activities on ultraslow-spreading ridges exhibit diverse characteristics, long histories with multiple participants, and might form large-scale, high-grade sulfide deposits. The Duanqiao hydrothermal field (DHF) is located at the segment with the thickest oceanic crust and a large axial magma chamber on the Southwest Indian Ridge, providing unique perspective of sulfide metallogenesis on ultraslow-spreading ridges. Previous studies revealed that DHF sulfide exhibits distinct features of enrichment of ore-forming elements in comparison with those of hydrothermal fields on sediment-starved mid-ocean ridges. However, the genesis and processes responsible for such differences remain poorly constrained. In this study, mineralogical, geochemical and S and Pb isotopic analyses were performed on relict sulfide mound samples to characterize DHF formation. The samples show clear concentric mineral zonation from the interior to the exterior wall. Assemblages of chalcopyrite, sphalerite, and pyrite are distributed mainly in the interior wall, whereas pyrite and marcasite are distributed mainly in the exterior wall. The low Cu content and Pb isotopic composition of the sulfide indicate that the metals are derived mainly from basement basalts. The δ34S values exhibit positive values distributed over a reasonably narrow range (2.42‰–7.97‰), which suggests approximately 62.1%–88.5% of S with basaltic origin. Compared with most hydrothermal fields along the sediment starved mid-ocean ridges, the DHF sulfide shows particularly high contents of Pb (263–2630 ppm), As (234–726 ppm), Sb (7.32–44.3 ppm), and Ag (35.2 to >100 ppm). The δ34S values exhibit an increasing tendency from the sample exterior to the interior. We propose that these features probably reflect the existence of a subsurface zone refining process. Our results provide new insight into the sulfide formation process and contribute to understanding the metallogenic mechanism of hydrothermal sulfides on ultraslow-spreading ridges.

  • Anthropogenic noise can be hazardous for the auditory system and wellbeing of animals, including humans. However, very limited information is known on how this global environmental pollutant affects auditory function and inner ear sensory receptors in early ontogeny. The zebrafish (Danio rerio) is a valuable model in hearing research, including investigations of developmental processes of the vertebrate inner ear. We tested the effects of chronic exposure to white noise in larval zebrafish on inner ear saccular sensitivity and morphology at 3 and 5 days post-fertilization (dpf), as well as on auditory-evoked swimming responses using the prepulse inhibition (PPI) paradigm at 5 dpf. Noise-exposed larvae showed a significant increase in microphonic potential thresholds at low frequencies, 100 and 200 Hz, while the PPI revealed a hypersensitization effect and a similar threshold shift at 200 Hz. Auditory sensitivity changes were accompanied by a decrease in saccular hair cell number and epithelium area. In aggregate, the results reveal noise-induced effects on inner ear structure–function in a larval fish paralleled by a decrease in auditory-evoked sensorimotor responses. More broadly, this study highlights the importance of investigating the impact of environmental noise on early development of sensory and behavioural responsiveness to acoustic stimuli.

  • Genotoxic effects of dicofol on the edible clam Meretrix meretrix were investigated through a mesocosm experiment. Individuals of M. meretrix, were exposed to environmental concentration (D1 = 50 ng/L) and supra-environmental concentration (D2 = 500 ng/L) of dicofol for 15 days, followed by the same depuration period. DNA damage (i.e., strand breaks and alkali-labile sites) was evaluated at day 1, 7 and 15, during uptake and depuration, using Comet assay (alkaline version) and nuclear abnormalities (NAs) as genotoxicity biomarkers. The protective effects of dicofol against DNA damage induced by ex vivo hydrogen peroxide (H2O2) exposure were also assessed. Comet assay results revealed no significant DNA damages under dicofol exposure, indicating 1) apparent lack of genotoxicity of dicofol to the tested conditions and/or 2) resistance of the animals due to optimal adaptation to stress conditions. Moreover, ex vivo H2O2 exposure showed an increase in the DNA damage in all the treatments without significant differences between them. However, considering only the DNA damage induced by H2O2 during uptake phase, D1 animals had significantly lower DNA damage than those from other treatments, revealing higher protection against a second stressor. NAs data showed a decrease in the % of cells with polymorphic, kidney shape, notched or lobbed nucleus, along the experiment. The combination of these results supports the idea that the clams used in the experiment were probably collected from a stressful environment (in this case Pearl River Delta region) which could have triggered some degree of adaptation to those environmental conditions, explaining the lack of DNA damages and highlighting the importance of organisms’ origin and the conditions that they were exposed during their lives.

  • The recently explored inactive Tianzuo hydrothermal field, in the amagmatic segment of the ultraslow-spreading Southwest Indian Ridge (SWIR), is closely associated with detachment faults. In this site, sulfide minerals are hosted by serpentine-bearing ultramafic rocks and include high-temperature (isocubanite, sphalerite, and minor pyrrhotite) and low-temperature (pyrite I, marcasite, pyrite II, and covellite) phases. In this study, trace-element concentrations of isocubanite and pyrite II were used to elucidate mineralization processes in ultramafic rocks hosting sulfides. Results show that isocubanite is enriched in metals such as Cu, Co, Sn, Te, Zn, Se, Pb, Bi, Cd, Ag, In, and Mn, and pyrite II is enriched in Mo and Tl. The marked enrichment in Te, Cu, Co, and In in isocubanite (compared with Se, Zn, Ni, and Sn, respectively) is most likely due to the contribution of magmatic fluids from gabbroic intrusions beneath the hydrothermal field. The intrusion of gabbroic magmas would have enhanced serpentinization reactions and provided a relatively oxidizing environment through the dissolution of anhydrite precipitated previously in the reaction zone, within high temperature and low pH conditions. This might have facilitated the extraction of metals by initial hydrothermal fluids, leading to the general enrichment of most metals in isocubanite. Metals in pyrite II have compositions similar to those of isocubanite, except for strong depletion in magmatically derived Te, Cu, Co, and In. This means that serpentinization processes had a dominating role in pyrite II precipitation as well. The enrichment of pyrite II in Mo and Tl is also indicative of seawater contribution in its composition. The study concludes that serpentinization reactions contribute effectively both to high- and low-temperature sulfide mineralization at Tianzuo hydrothermal field, with gabbroic intrusions further promoting high-temperature sulfide mineralization, providing additional metals, fluids and heat. In contrast, low-temperature sulfide mineralization occurred during the cooling of gabbroic intrusions, with decreasing rates of serpentinization reactions and a significant influence of seawater.

  • Mangrove forests are one of the most ecologically valuable ecosystems in the world and provide a wide variety of ecosystem services to coastal communities, including cities. Macao, a highly urbanized coastal city located on the southern coast of China west of the Pearl River, is home to several species of mangroves with many associated flora and fauna. Mangrove forests in Macao are vulnerable to threats due to pressure from rapid and massive urban developments in the area, which led to mangrove loss in the past decades. To address this issue, the local authorities established special Ecological Zones for the management of the local mangroves. To reinforce local conservation efforts, educating the local population about the value of mangroves, especially school students, is of utmost importance. To evaluate the impact of environmental education activities on the environmental orientation, knowledge, and values of students toward mangrove conservation in Macao, a quasi-experimental study was undertaken. The effectiveness of a mangroves exhibition and field visit were evaluated using the New Environmental Paradigm (NEP) Scale—Macao version in a group of local school students who participated in the activities. Overall, the results provided consistently positive evaluations of the impact of the environmental education program. The strongest improvements were found in the students’ pro-environmental orientations, knowledge about mangroves, and value for environmental protection.

  • AimsVascular calcification is a common clinical complication of chronic kidney disease (CKD), atherosclerosis (AS), and diabetes, which is associated with increased cardiovascular morbidity and mortality in patients. The transdifferentiation of vascular smooth muscle cells (VSMCs) to an osteochondrogenic phenotype is a crucial step during vascular calcification. The transcription factor CCAAT/enhancer-binding protein alpha (C/EBPα) plays an important role in regulating cell proliferation and differentiation, but whether it regulates the calcification of arteries and VSMCs remains unclear. Therefore, this study aims to understand the role of C/EBPα in the regulation of vascular calcification.Methods and ResultsBoth mRNA and protein expression levels of C/EBPα were significantly increased in calcified arteries from mice treated with a high dose of vitamin D3 (vD3). Upregulation of C/EBPα was also observed in the high phosphate- and calcium-induced VSMC calcification process. The siRNA-mediated knockdown of C/EBPα significantly attenuated VSMC calcification in vitro. Moreover, C/EBPα depletion in VSMCs significantly reduced the mRNA expression of the osteochondrogenic genes, e.g., sex-determining region Y-box 9 (Sox9). C/EBPα overexpression can induce SOX9 overexpression. Similar changes in the protein expression of SOX9 were also observed in VSMCs after C/EBPα depletion or overexpression. In addition, silencing of Sox9 expression significantly inhibited the phosphate- and calcium-induced VSMC calcification in vitro.ConclusionFindings in this study indicate that C/EBPα is a key regulator of the osteochondrogenic transdifferentiation of VSMCs and vascular calcification, which may represent a novel therapeutic target for vascular calcification.

  • In southeast Asia, males of the Siamese fighting fish, Betta splendens, have been selected across centuries for winning paired staged fights and previous work has shown that males from fighter strains are more aggressive than wild-types. This strong directional selection for winners is likely to have targeted aggression-related endocrine systems, and a comparison between fighter and wild-type strains can bring into evidence the key hormones implicated in aggression. Here, we compared the plasma levels of the androgen 11-ketotestosterone (KT) and of the corticosteroid cortisol (F) in F2 males of a fighter and a wild-type strain raised under similar laboratory conditions. We show that F was generally lower in fighter as compared with wild-type males, while no overall differences in KT levels were detected between strains. When presented with a mirror-induced aggressive challenge, post-fight levels of F increased but more significantly so in wild-type males, while KT increased in males of both strains. After the challenge, fighter males had higher levels of KT as compared with wild-type males, while the pattern for F was opposite. As compared with animals in social groups, wild-type males placed under social isolation had lower F levels, while KT decreased for fighters. Taken together, this data suggests that while wild-type males responded to aggression with an increase in circulating levels of both androgens and corticosteroids, males selected for winning fights maintained a blunt F response, increasing only KT levels. These data agree with the hypothesis that a combination of high levels of androgens and low levels of corticosteroids is associated with high aggression. Overall, these results seem to indicate that selection for winning had a stronger impact in the hypothalamus-pituitary-interrenal axis than in the hypothalamus-pituitary–gonadal axis in B. splendens.

  • Environmental education (EE) has long been practiced worldwide, while Nature-based solutions (NBS) is a relatively new concept. This chapter aims to provide an overview of the EE and NBS practices in East Asia and evaluate how these two valuable applications can be used concurrently. East Asia has a well developed environmental education (EE) programs and activities, both in formal and informal education. These ranges from developing green schools and campuses to establishing policies and acts. While EE has been actively practiced for decades in the region, the adoption of NBS to address environmental and societal challenges is limited. The educational benefits and opportunities from NBS are also lacking. Although there are some projects that can be classified as NBS, like the use of wetlands for wastewater treatment, they are not clearly categorized as one. These projects are also not integrated into environmental education programs. Considering this, the region should develop innovative environmental education programs for schools, universities and communities, that integrate NBS projects. Integrating the two together will boost the effectiveness of environmental education in raising environmental awareness and changing the environmental attitude and behavior of people, which will also help address societal issues.

  • As safety is one of the most important properties of drugs, chemical toxicology prediction has received increasing attentions in the drug discovery research. Traditionally, researchers rely on in vitro and in vivo experiments to test the toxicity of chemical compounds. However, not only are these experiments time consuming and costly, but experiments that involve animal testing are increasingly subject to ethical concerns. While traditional machine learning (ML) methods have been used in the field with some success, the limited availability of annotated toxicity data is the major hurdle for further improving model performance. Inspired by the success of semi-supervised learning (SSL) algorithms, we propose a Graph Convolution Neural Network (GCN) to predict chemical toxicity and trained the network by the Mean Teacher (MT) SSL algorithm. Using the Tox21 data, our optimal SSL-GCN models for predicting the twelve toxicological endpoints achieve an average ROC-AUC score of 0.757 in the test set, which is a 6% improvement over GCN models trained by supervised learning and conventional ML methods. Our SSL-GCN models also exhibit superior performance when compared to models constructed using the built-in DeepChem ML methods. This study demonstrates that SSL can increase the prediction power of models by learning from unannotated data. The optimal unannotated to annotated data ratio ranges between 1:1 and 4:1. This study demonstrates the success of SSL in chemical toxicity prediction; the same technique is expected to be beneficial to other chemical property prediction tasks by utilizing existing large chemical databases. Our optimal model SSL-GCN is hosted on an online server accessible through: https://app.cbbio.online/ssl-gcn/home.

  • Microbial and hydrothermal venting activities on the seafloor are important for the formation of sediment-hosted stratiform sulfide (SHSS) deposits. Fe isotopic compositions are sensitive to both microbial and hydrothermal activities and may be used to investigate the formation of these deposits. However, to the best of our knowledge, no Fe isotopic studies have been conducted on SHSS deposits. In the Devonian Dajiangping SHSS-type pyrite deposit (389 Ma), South China, laminated pyrite ores were precipitated from exhalative hydrothermal fluids, whereas black shales were deposited during intervals with no exhalation. Pyrite grains from black shales mostly display positive δ56Fe-py (0.01–0.73‰), higher than marine sediments (ca. 0‰), due to pyrite deriving Fe from basinal shuttled Fe(III) (hydr-)oxides and slowly crystallizing in pores of sediments with equilibrium fractionation, except for negative δ56Fe-py (−0.17‰ to −0.24‰) of two samples caused by mixing of Fe from underlain laminated ores. The positive δ34S-py (3.50–24.5‰) of black shales reflect that sulfur of pyrite originated from quantitative reduction of sulfate in closed pores of sediments. In contrast, pyrite grains of laminated ores have negative δ56Fe-py (−0.60‰ to −0.21‰), which were not only inherited from the negative δ56Fe of hydrothermal fluids but also caused by kinetic fractionation during rapid precipitation of a pyrite precursor (FeS) in hydrothermal plumes. These ores have negative δ34S-py (−28.7‰ to −1.82‰), because H2S for pyrite mineralization was produced by bacterial sulfate reduction (BSR) in a sulfate-rich seawater column or shallow sediments. The δ56Fe-py values of laminated ores co-vary positively with δ34S-py and δ13C-carbonate along the ore stratigraphy, with δ13C-carbonate values ranging from −12.0‰ to −2.50‰. However, they correlate negatively with aluminum-normalized total organic carbon (TOC/Al2O3). Organic carbon is thus considered to enhance the production of H2S by BSR activities, increase pyrite precipitation rates and promote the expression of kinetic fractionation of Fe isotopes. Intriguingly, in the ore units with vigorous hydrothermal venting activities, δ56Fe-py, δ34S-py and δ13C-carbonate values display a consistently increasing trend. Such results suggest that venting hydrothermal fluids significantly inhibited the H2S production of BSR, which then reduced the pyrite crystallization rate and decreased the kinetic fractionation of Fe isotopes. Our study reveals that the formation of SHSS deposits relies on H2S from microbial activities and metals from hydrothermal exhalation on the seafloor, but that vigorous exhalation can inhibit microbial activities and thus sulfide precipitation rates. The integrated use of Fe, S, and C isotopes can effectively elucidate these dynamic interactions between hydrothermal venting and microbial activities during the formation of SHSS deposits.

  • The occurrence of endocrine disrupting chemicals (EDCs) is a major issue for marine and coastal environments in the proximity of urban areas. The occurrence of EDCs in the Pearl River Delta region is well documented but specific data related to Macao is unavailable. The levels of bisphenol-A (BPA), estrone (E1), 17α-estradiol (αE2), 17β-estradiol (E2), estriol (E3), and 17α-ethynylestradiol (EE2) were measured in sediment samples collected along the coastline of Macao. BPA was found in all 45 collected samples with lower BPA concentrations associated to the presence of mangrove trees. Biodegradation assays were performed to evaluate the capacity of the microbial communities of the surveyed ecosystems to degrade BPA and its analogue BPS. Using sediments collected at a WWTP discharge point as inoculum, at a concentration of 2 mg l−1 complete removal of BPA was observed within 6 days, whereas for the same concentration BPS removal was of 95% after 10 days, which is particularly interesting since this compound is considered recalcitrant to biodegradation and likely to accumulate in the environment. Supplementation with BPA improved the degradation of bisphenol-S (BPS). Aiming at the isolation of EDCs-degrading bacteria, enrichments were established with sediments supplied with BPA, BPS, E2 and EE2, which led to the isolation of a bacterial strain, identified as Rhodoccoccus sp. ED55, able to degrade the four compounds at different extents. The isolated strain represents a valuable candidate for bioremediation of contaminated soils and waters.

  • The Dayingezhuang gold deposit in the Jiaodong district, eastern margin of the North China Craton is hosted in Mesozoic granitic rocks and consists of quartz-sulfide veins/veinlets and sulfide disseminations in alteration envelopes. Previous studies mainly focused on the geochronology, sources of ore-forming fluids and metals to investigate the ore genesis. However, enrichment mechanism of Au and other associated trace metals remain unclear. In this study, we present detailed textures and in-situ LA-ICP-MS trace-element compositions of different generations of pyrite, as well as EMP analysis of Au-bearing minerals to discuss the occurrence and enrichment mechanism of Au at this deposit. Three generations of pyrite (Py1, Py2 and Py3) formed during three hydrothermal ore stages (I, II, and III) at Dayingezhuang. Py1 occurs as disseminations in sericitic alteration assemblages and is characterized by low Au (mean 0.15 ppm), Ag, As and Te contents. The time-resolved depth-concentration profiles indicate that Au in Py1 mainly occurs as nanoparticles and/or micron-sized inclusions. Py2 can be further divided into the early undeformed Py2a and later Py2b, which is the product of deformed Py2a with different degrees of brittle to plastic deformation and recrystallization. Py2a in pyrite-siderite-quartz veins is relatively enriched in invisible Au (mean 0.41 ppm), Ag, As, and Te compared to Py1, and contains numerous micron-sized Au inclusions. In contrast, Py2b contains lesser invisible Au (0.21 ppm) and host abundant gold minerals along the grain boundaries and microfractures. Py3 in polymetallic sulfide veins has little Au. As a whole, Au in pyrite is positively correlated with Ag and Te, which is consistent with the results of EMP analysis showing the occurrence of Au as electrum, native gold and minor petzite in pyrite. Such evidences show that the deformation and recrystallization of auriferous Py2a potentially caused local remobilization of Au (mainly as micron-sized inclusion Au) via solid-state ductile flow and subsequent reconcentration of Au in microfractures of Py2b. The pyrite deformation and Au remobilization events were suggested to be related to the continuous reactivation of the regional Zhaoping Fault contemporaneous with gold mineralization. Our study highlights the importance of remobilization and reconcentration of Au triggered by syn-ore tectonic activities at Dayingezhuang and possibly other Au deposits in the Jiaodong district.

  • The role of hormones as modulators of aggressive behavior in fish remains poorly understood. Androgens and corticosteroids, in particular, have been associated with aggressive behavior in fish but it is still not clear if animals adjust the secretion of these hormones to regulate behavior during ongoing fights, in response to fight outcomes in order to adjust aggressive behavior in subsequent fights, or both. With its stereotyped displays and high aggression levels, the Siamese fighting fish Betta splendens is an excellent model to investigate this question. Here, we compared the behavioral and endocrine response of male B. splendens to fights where there is no winner or loser by presenting them with a size-matched live interacting conspecific behind a transparent partition or with a mirror image. The aggressive response started with threat displays that were overall similar in frequency and duration towards both types of stimuli. Fights transitioned to overt attacks and interacting with a live conspecific elicited a higher frequency of attempted bites and head hits, as compared with the mirror image. There was a pronounced increase in plasma androgens (11-ketotestosterone and testosterone) and corticosteroids (cortisol) levels in response to the aggression challenge, independent of stimulus type. Post-fight intra-group levels of these hormones did not correlate with measures of physical activity or aggressive behavior. A linear discriminant analysis including all behavioral and endocrine data was a poor classifier of fish from the conspecific and mirror trials, showing that overall the behavioral and endocrine response to mirror images and conspecifics was similar. The results show that fight resolution is not necessary to induce an evident increase in peripheral levels of androgens and corticosteroids in B. splendens. However, the function of these hormones during present and future aggressive contests remains to be clarified.

  • The Qiyugou gold deposit, located in the Xiong’ ershan area of the North China Craton, contains abundant bismuth-sulfosalts that are closely associated with gold mineralization. Pyrite is the dominant Au-hosted mineral, and has been formed in three generations (Py1, Py2, and Py3). Py1 grains, generally intergrown with milky quartz, are coarse (>1 mm), euhedral in shape, and Au-depleted in composition. In contrast, subhedral Py2 grains, associated with light gray quartz, are medium to coarse (0.2–3 mm) and are enriched in gold that is both invisible and visible. Py3 grains (0.1–0.5 mm), intergrown with abundant sulfide minerals, are relatively fine and Au-depleted. The time-resolved LA-ICP-MS depth profiles of the Py2 grains indicate that invisible gold occurs as either solid solution or nano-particles of native gold and electrum. Visible gold occurs as small blebs in the Py2 grains where inclusions of native bismuth, galenobismutite, lillianite homologs, tetradymite, and galena are also present. In addition, it is common that electrum in microfracture infillings or along grain boundaries of the Py1 and Py2, are intergrown with bismuthinite derivatives, Bi-Cu sulfosalts, emplectite, tetradymite, chalcopyrite, galena, and Py3. Based on textural relationships and mineral assemblages, calculation of physicochemical conditions show that gold was formed in conditions of fTe2 = ~10−11 and fS2 = ~10−11 to 10−12 for Py2, and fTe2 = ~10−9 to 10-11and fS2 = ~10−10 to 10−11 for Py3. We thus proposed that such physicochemical conditions may have triggered the precipitation of Bi melt, and sulfidation driven by cooling or increase in sulfur content results in the transformation of the Au-Bi liquid into a stable assemblage of native gold and bismuthinite. These bismuth minerals are associated with native gold/Au-bearing minerals, indicating that the Au mineralization of the Qiyugou gold deposit might be genetically associated with Bi melt. The present study highlights the role of Bi as important gold scavengers in arsenic-deficient ore-forming fluid.

  • Mangroves are a unique group of plants growing along tropical and sub-tropical coastlines, with the ability to remove several types of contaminants such as heavy metals and other persistent organic compounds in coastal waters. However, little attention has been given to the possible role of mangroves in the removal of organochlorinated pesticides (OCPs) from the environment. Used worldwide, these pesticides were banned in the late 80s, withal they can still be quantified in aquatic environments due to their high stability. Moreover, as persistent and lipophilic compounds, OCPs are known for their tendency to bioaccumulate and biomagnify through the food chain, affecting local ecosystems, and potentially human health. This work aimed to investigate the potential benefits of mangrove ecosystems as OCP phytoremediators. For this purpose, a total of seventy-three articles from non-mangrove and mangrove areas around the world were gathered, integrated and re-analysed as a whole. These data include information from four different matrices (water, sediment, benthic fauna and mangrove plants). A common trend of less pesticide contamination in mangrove areas was observed for all the selected matrices. As a complement, average concentrations were discussed considering International Directives, such as the European legislation 2013/39/EU for water policy and the Dutch List together with the International Sediment Quality Guideline, for sediments. Additionally, theoretical risk assessments were also included. Since information regarding OCPs in mangroves ecosystem is very scarce compared to non-mangrove areas, this review provides valuable insights regarding these environments, and the importance of preserving them as a relevant remediation unit.

  • Seafloor massive sulfide (SMS) deposits are important deep-sea mineral resources expected to occur predominantly on slow- and ultraslow-spreading mid-ocean ridges. Resource estimates are already available for some of the largest SMS deposits on slow-spreading ridges but not on ultraslow-spreading ridges. Based on geological mapping and sampling, this study investigates the distribution and content of sulfide-rich deposits in the Yuhuang-1 hydrothermal field (YHF), located on the ultraslow-spreading Southwest Indian Ridge. The sulfide-rich deposits in the YHF are composed of two areas ∼500 m apart: the southwest sulfide area (SWS) and the northeast sulfide area (NES). We calculated the volume of sulfide-rich mounds in the YHF and arrived at a total accumulation of ∼10.6 × 106 tons, including at least ∼7.5 × 105 tons of copper and zinc and ∼18 tons of gold. Furthermore, considering the coverage of layered hydrothermal sediment mixed with sulfide-rich breccias, which may have underlying massive sulfide deposits, the maximum total mass was estimated at ∼45.1 × 106 tons. This suggests that the YHF is one of the largest SMS deposits worldwide and confirm that ultraslow-spreading ridges have the greatest potential to form large-scale SMS deposits.

Last update from database: 3/28/24, 7:44 PM (UTC)