[Association between polymorphism of CASP and NOX3 with risk of noise-induced hearing loss].

Objective: To explore the effect of gene polymorphism on workers suffering from noise induced hearing loss (NIHL) . Methods: In May 2019, a case-control study was conducted to select noise exposed workers in five factories in Zhejiang Province from 2017 to 2018. The average hearing threshold of binaural high frequency (3, 4, 6 kHz) was >25 dB (A) as the NIHL group, and the hearing threshold of any language frequency (0.5, 1, 2 kHz) was ≤25 dB (A) as the non NIHL group, with 307 people in each group. The general demographic data, occupational history, pure tone audiometry results and oral swab mucosal samples of noise exposed workers were collected, and the DNA of oral mucosal cells was extracted. The relationship between genetic risk score (GRS) and NIHL was analyzed, single nucleotide polymorphisms (SNP) were genotyped, the relationship between genotype and NIHL was analyzed by logistic regression, and the relationship between haplotype and NIHL was analyzed by R language. Results: After adjusting for gender, age, education and working years, the risk of NIHL among workers carrying cysteine-aspartic acid protease 3 gene (CASP3) rs1049216 recessive model GG genotype, rs6948 recessive model TT genotype, NADPH oxidase 3 gene (NOX3) rs12195525 additive model GT genotype and dominant model TT+GT genotype decreased (P<0.05) , the risk of disease was higher in workers with AA genotype carrying cysteine-aspartic acid protease 7 gene (CASP7) rs12415607 additive model (P<0.05) . There was a strong linkage disequilibrium (LD) relationship between rs1049216 and rs6948 (D'>0.8) . Haplotype AT and GG composed of rs1049216-rs6948 increased the risk of NIHL (P<0.05) . The risk of NIHL increased with the increase of GRS (OR=2.69, P<0.05) . Conclusion: Genotype polymorphisms at rs1049216 and rs6948 (CASP3) , rs12195525 (NOX3) , rs12415607 (CASP7) may be associated with susceptibility to NIHL.

Myocardial CT perfusion imaging and atherosclerotic plaque characteristics on coronary CT angiography for the identification of myocardial ischaemia.

AIM: To investigate the value of myocardial computed tomography (CT) perfusion imaging (CTP) and atherosclerotic plaque characteristics (APCs) identified on coronary CT angiography (CCTA) for the detection of myocardial ischaemia by using single-photon-emission CT (SPECT) as a reference. MATERIALS AND METHODS: Thirty-six patients (63.9% males) undergoing combined stress dynamic CTP and CCTA were enrolled and analysed. Myocardial blood flow (MBF) from CTP was quantified and compared between normal and abnormal segments. The ability of CTP and APCs to detect ischaemia was compared to that of SPECT. RESULTS: Nineteen patients with 78 segments had perfusion abnormalities on CTP. A significant difference was seen in MBF values between normal (118.51±27.86 ml/100 ml/min) and hypoperfused (79.60±21.35 ml/100 ml/min) segments (t=15.832, p<0.05). The sensitivity and specificity for identifying ischaemia were 90.91% and 94.97%, respectively, on a per-segment basis, resulting in a r value of 0.737 (p<0.05). On a per-vessel basis, the sensitivity and specificity for detecting ischaemia were 86.67% and 84.62%, respectively, for CTP; 93.33% and 58.97%, respectively, for CCTA; and 86.67% and 87.18%, respectively, for CTP combined with CTA, with an area under the receiver-operator characteristic curve (AUC) being 0.87 (p<0.05) and 0.887 (p<0.05) for CTP and its combination with CTA, respectively. On CCTA, 55 vessels with APCs were detected, with an AUC of 0.737 (p<0.05) for APCs combined with CCTA stenosis and 0.802 (p<0.05) for APCs combined with CTP. CONCLUSIONS: Dynamic stress CTP shows good correlation with SPECT for the detection of ischaemia. Additionally, combining APCs with CCTA stenosis has the ability to discriminate ischaemic stenosis.

Interactive effects of hypoxia and PBDE on larval settlement of a marine benthic polychaete.

Marine benthic polychaete Capitella sp. I is widely known to adapt to polluted habitats; however, its response to xenobiotics under hypoxic conditions has been rarely studied. This research aimed to test the hypothesis that interactive effects of hypoxia and congener BDE-47 of polybrominated diphenyl ethers (PBDE), which is ubiquitous in marine sediments, may alter the settlement of Capitella sp. I. Our results revealed that under hypoxic condition, settlement success and growth in body length of Capitella larvae were significantly reduced compared to those under normoxia of similar BDE-47 concentration. While no significant changes in morphology of settled larvae were noted in both exposure conditions, the presence of BDE-47 could enhance polychaete growth. The present findings demonstrated that the interactive effects of hypoxia and environmentally realistic concentrations of BDE-47 in sediments could affect polychaete settlement, which, in turn, reduce its recruitment and subsequent population size in the marine benthic ecosystem.

Early detection of acute radiation-induced lung injury with multi-section CT perfusion imaging: An initial experience.

AIM: To explore the value of 64-section computed tomography (CT) perfusion imaging (CTPI) in the early diagnosis of acute radiation-induced lung injury (ARILI). MATERIALS AND METHODS: Fifty-one patients with oesophageal cancers or malignant thymomas received postoperative radiation therapy with a 60-62 Gy dose and underwent CTPI at pre- and post-radiation therapy time points (week 0, week 4, week 8, and week 12 respectively). The CTPI values were prospectively compared and analysed in order to evaluate the diagnostic utility of CTPI in the early diagnosis of ARILI. RESULTS: Eighteen cases (18/51) of ARILI were diagnosed. The mean values of relative regional blood flow (rrBF), relative regional volume (rrBV), and relative regional permeability surface (rrPS) in the ARILI group were correspondingly higher than those of the non-ARILI group. At week 4, rrBF, rrBV, and rrPS in the ARILI group were significantly higher than those at pre-radiation (each p < 0.05). In the non-ARILI group, rrBF and rrBV were higher than those at pre-radiation (each p < 0.05); however, rrPS was not statistically different from that of pre-irradiation. Applying the diagnostic threshold value of rrPS = 1.22, the sensitivity, specificity, and positive and negative predictive values of CTPI for early diagnosis of ARILI were better than those of CT. CONCLUSION: CTPI metrics may reflect haemodynamic changes in the post-irradiation lung and can detect cases of early ARILI that appear normal at CT. CTPI is a promising technique for early diagnosis of ARILI.

Distinctive microbial community structure in highly stratified deep-sea brine water columns.

Atlantis II and Discovery are two hydrothermal and hypersaline deep-sea pools in the Red Sea rift that are characterized by strong thermohalo-stratification and temperatures steadily peaking near the bottom. We conducted comprehensive vertical profiling of the microbial populations in both pools and highlighted the influential environmental factors. Pyrosequencing of the 16S rRNA genes revealed shifts in community structures vis-à-vis depth. High diversity and low abundance were features of the deepest convective layers despite the low cell density. Surprisingly, the brine interfaces had significantly higher cell counts than the overlying deep-sea water, yet they were lowest in diversity. Vertical stratification of the bacterial populations was apparent as we moved from the Alphaproteobacteria-dominated deep sea to the Planctomycetaceae- or Deferribacteres-dominated interfaces to the Gammaproteobacteria-dominated brine layers. Archaeal marine group I was dominant in the deep-sea water and interfaces, while several euryarchaeotic groups increased in the brine. Across sites, microbial phylotypes and abundances varied substantially in the brine interface of Discovery compared with Atlantis II, despite the near-identical populations in the overlying deep-sea waters. The lowest convective layers harbored interestingly similar microbial communities, even though temperature and heavy metal concentrations were very different. Multivariate analysis indicated that temperature and salinity were the major influences shaping the communities. The harsh conditions and the low-abundance phylotypes could explain the observed correlation in the brine pools.

De novo assembly of the transcriptome of an invasive snail and its multiple ecological applications.

Studying how invasive species respond to environmental stress at the molecular level can help us assess their impact and predict their range expansion. Development of markers of genetic polymorphism can help us reconstruct their invasive route. However, to conduct such studies requires the presence of substantial amount of genomic resources. This study aimed to generate and characterize genomic resources using high throughput transcriptome sequencing for Pomacea canaliculata, a nonmodel gastropod indigenous to Argentina that has invaded Asia, Hawaii and southern United States. De novo assembly of the transcriptome resulted in 128,436 unigenes with an average length of 419 bp (range: 150-8556 bp). Many of the unigenes (2439) contained transposable elements, showing the existence of a source of genetic variability in response to stressful conditions. A total of 3196 microsatellites were detected in the transcriptome; among 20 of the randomly tested microsatellites, 10 were validated to exhibit polymorphism. A total of 15,412 single-nucleotide polymorphisms (SNPs) were detected in the ORFs. LC-MS/MS analysis of the proteome of juveniles revealed 878 proteins, of which many are stress related. This study has demonstrated the great potential of high throughput DNA sequencing for rapid development of genomic resources for a nonmodel organism. Such resources can facilitate various molecular ecological studies, such as stress physiology and range expansion.

Polybrominated diphenylethers (PBDEs) alter larval settlement of marine benthic polychaetes.

Polybrominated diphenylethers (PBDEs) are found ubiquitously in marine environments worldwide. Sediment is the major sink of PBDEs, with the congener BDE 47 being most abundant. In this study, laboratory experiments were carried out to test the hypothesis that contamination of BDE 47 at environmentally realistic sediment concentrations can alter polychaete larval settlement. Using multiple-choice experiment, settlement of three polychaete species (Pseudopolydora vexillosa, Polydora cornuta, and Capitella sp. I) on four types of spiked sediment was studied and compared: (i) low BDE 47 concentration (0.5 ng g(-1) dry weight); (ii) high BDE 47 concentration (3.0 ng g(-1) dry weight), (iii) hexane (solvent control), and (iv) natural sediment (control). Our results showed that settlement of P. vexillosa and Capitella sp. I larvae was significantly promoted, while settlement of P. cornuta reduced, at high BDE 47 concentration in sediment compared with the respective controls under both short- (24 h) and long-term (4 week) exposures. After 4 weeks, body burden of BDE 47 in all polychaete species was directly related to the spike concentration, and body length of settled juveniles of P. vexillosa and Capitella sp. I at the high-concentration treatment was significantly longer compared with that of other treatments and controls. For the first time, we demonstrated that environmentally realistic concentrations of BDE 47 in sediment can affect polychaete settlement in species-specific and dose-dependent manners. Given the global contamination of PBDE in marine sediment, BDE 47 may potentially alter the settlement pattern of marine polychaetes and hence their benthic composition over large areas.

Temporal variation of coastal surface sediment bacterial communities along an environmental pollution gradient.

Terminal restriction fragment length polymorphism analysis (T-RFLP) was used to track the changes of bacterial community compositions (BCC) in coastal surface sediments along an environmental pollution gradient between 2004 and 2006. BCC in the chronically contaminated sites showed the largest deviation from those in the adjacent sites. Surprisingly, BCC at two contrasting environments (oceanic vs. river-influenced) were more similar. Unexpectedly, the BCC did not recover (when compared to oceanic control site) even after 5 years of pollution abatement initiatives in Victoria Harbour, Hong Kong. On the other hand, disposal of treated sewage for 5 years in one of the sites did not significantly affect the BCC. A striking seasonal variation in the BCC was observed at only the polluted sites. Although factors other than pollution gradients may explain the observed BCC patterns, the information presented here can be useful in predicting long-term effects of pollution on BCC. Furthermore, this study suggests that BCC analysis using T-RFLP is a faster, reliable and easier approach to monitor microbenthic community response to environmental pollution gradient in coastal sediments.

Antifouling and antibacterial compounds from the gorgonians Subergorgia suberosa and Scripearia gracillis.

In this study, we investigated the potential antilarval and antibacterial activity of secondary metabolites of the gorgonians Subergorgia suberosa and Scripearia gracillis from the South China Sea. Fresh specimens of these two gorgonian corals were collected from a shallow reef in Sanya Bay of Hainan Island and extracted with different solvents. Antilarval activity of the chemical extracts and pure compounds was evaluated in settlement inhibition assays with laboratory-reared Balanus amphitrite and Bugula neritina larvae, while antibacterial activity was assessed with disc diffusion bioassay on growth inhibition of 15 marine bacterial species. Using bioassay-guided procedures, we purified and identified nine compounds. The most potent metabolites produced by these gorgonian corals were subergorgic acid and pregn-4-ene-3, 20-dione extracted from S. suberosa. Our results show that the gorgonian coral S. suberosa and S. gracillis can produce potent anti-fouling compounds that deserve further exploration.

Spatio-temporal changes of marine macrobenthic community in sub-tropical waters upon recovery from eutrophication. I. Sediment quality and community structure.

A two-year study was conducted in the vicinity of a harbour in sub-tropical Hong Kong, to examine the progress of improvement in sediment quality and recovery of macrobenthic community upon cessation of organic pollution caused by sewage discharge. Median sediment particle diameter (Mdphi) and levels of total organic carbon (TOC), total Kjeldahl nitrogen (TKN), ammonia-nitrogen (NH(3)-N) and total phosphorus (TP), as well as macrobenthic species composition and abundance were determined bi-monthly at three inside-harbour and two outside-harbour locations. At the two inside-harbour locations, significantly higher levels of TOC, TKN, NH(3)-N and TP in sediments were observed than the outside-harbour locations. However, no significant temporal change of nutrient levels was detected at all sampling locations during the two-year study, except a significant decrease in TKN and NH(3)-N levels at one outside-harbour location and a decline in NH(3)-N content at another outside-harbour location. Spatially, the highest in mean total species number, individual number, species diversity H' and lowest evenness J was found at one outside-harbour location, whereas the other four locations had relatively similar values. H' was negatively correlated with TOC, TKN, NH(3)-N and TP content in sediments while J was positively correlated with MDphi. Across the study period, the patterns of macrobenthic community were significantly different among all samplings and that the spatial difference of benthic community was best correlated with MDphi, TOC, TKN and water depth. A weak sign of recovery in macrobenthic community structure was detected at inside-harbour locations, with replacement of opportunistic by ubiquitous species over the two-year study. The slow recovery of macrobenthic community was related to sediment characteristics. Results of a larval settlement bioassay using the polychaete Capitella sp. I also demonstrated that the inside-harbour sediments were still unfavourable for colonization and larval settlement of species sensitive to pollution. The slow biodegradation of the organic pollutants and continuous presence of heavy metals in the sediment may hinder settlement and colonization of benthic animals. However, increases of H' and J were observed in a longer time scale when comparing the present data with those obtained four years ago. This suggested that a detectable recovery of benthic community in the harbour may take at least three years and a complete recovery may even take longer duration.

Effect of agitation on violacein production in Pseudoalteromonas luteoviolacea isolated from a marine sponge.

AIMS: Experiments were designed to investigate the effect of agitation on the production of violacein by a marine bacterium Pseudoalteromonas luteoviolacea. METHODS AND RESULTS: A marine sponge-associated bacterium, P. luteoviolacea, was grown at different agitation speeds. Agitation did not have a significant effect on bacterial growth, but had a profound effect on the size of bacterial aggregate. The production of violacein was the highest under stagnant conditions and decreased with the increase of the agitation speed. CONCLUSIONS: Agitation affected the aggregation of bacterial cells, which, in turn, affected violacein production by P. luteoviolacea. SIGNIFICANCE AND IMPACT OF THE STUDY: This study suggests that P. luteoviolacea produced the highest amount of violacein when it was cultured under stagnant conditions.

Marine biofilms as mediators of colonization by marine macroorganisms: implications for antifouling and aquaculture.

In the marine environment, biofilms on submerged surfaces can promote or discourage the settlement of invertebrate larvae and macroalgal spores. The settlement-mediating effects of biofilms are believed to involve a variety of biofilm attributes including surface chemistry, micro-topography, and a wide range of microbial products from small-molecule metabolites to high-molecular weight extracellular polymers. The settled organisms in turn can modify microbial species composition of biofilms and thus change the biofilm properties and dynamics. A better understanding of biofilm dynamics and chemical signals released and/or stored by biofilms will facilitate the development of antifouling and mariculture technologies. This review provides a brief account of 1) existing knowledge of marine biofilms that are relevant to settlement mediation, 2) biotechnological application of biofilms with respect to developing non-toxic antifouling technologies and improving the operation of aquaculture facilities, and 3) challenges and future directions for advancing our understanding of settlement-mediating functions of biofilms and for applying this knowledge to real-life situations.

Interactions of rice (Oryza sativa L.) and PAH-degrading bacteria (Acinetobacter sp.) on enhanced dissipation of spiked phenanthrene and pyrene in waterlogged soil.

The effects of cultivation of rice (Oryza sativa L.) and PAH-degrading bacteria (Acinetobacter sp.) separately, and in combination, on the dissipation of spiked phenanthrene and pyrene (0, 50+50, 100+100, 200+200 mg kg(-1)) in waterlogged soil were studied using pot trials. The population of introduced PAH-degrading bacteria remained at 10(5) CFU g(-1) dry soil after 20 days of treatment with Acinetobacter sp. only, but increased to 10(6) when planted with rice simultaneously. Shoot and root biomass of rice when grown alone was adversely affected by spiked PAHs, but significantly increased by 2-55% and 8-409%, respectively, when inoculated with Acinetobacter sp.. Phenanthrene and pyrene concentrations in roots ranged from 1-27 and 20-98 mg kg(-1), respectively, while their concentrations in shoots were generally lower than 0.2 mg kg(-1). The dissipation of phenanthrene was mainly due to abiotic loss as 70-78% phenanthrene was lost from the control soil at the end of 80 days, while removal of 86-87% phenanthrene had been achieved after 40 days in the treatment co-cultivated with Acinetobacter sp. and rice. Compared with the control where only 6-15% of pyrene was removed from soil, a much higher dissipation of pyrene (43-62%) was attained for the treatments co-cultivated with Acinetobacter sp. and rice at the end of 80 days. The results demonstrated that co-cultivation of rice and PAH-degrading bacteria may have a great potential to accelerate the bioremediation process of PAH-contaminated soil under waterlogged conditions.

Antifouling properties of 10beta-formamidokalihinol-A and kalihinol A isolated from the marine sponge Acanthella cavernosa.

Many soft-bodied sessile marine invertebrates such as sponges and soft corals defend themselves against fouling directly through the production of antifouling compounds, or indirectly through regulating the epibiotic microbes that affect larval settlement. In this study, 10beta-formamidokalihinol-A and kalihinol A were isolated and purified from the marine sponge Acanthella cavernosa (Dendy). The results indicated that both compounds inhibited the growth of bacteria isolated from the natural environment whereas kalihinol A suppressed larval settlement of a major fouling polychaete, Hydroides elegans with an EC50 of 0.5 microg ml(-1). Kalihinol A was incorporated in Phytagel that was exposed to the bacterial consortia in natural seawater for biofilm formation. Biofilms that developed on the Phytagel surfaces were analysed for bacterial abundance and bacterial species composition using a DNA fingerprinting technique, terminal restriction fragment length polymorphism (T-RFLP). The results showed that kalihinol A only slightly reduced bacterial abundance (t-test, p = 0.0497), but modified the bacterial species composition of the biofilms. Inhibition of H. elegans larval settlement was observed when biofilms developed under the influence of kalihinol A were exposed to larvae, suggesting that compounds like kalihinol A from the sponge A. cavernosa may change bacterial community composition on the sponge surface, which in turn, modulates larval settlement of fouling organisms.

Long-term changes in water quality and phytoplankton characteristics in port shelter, Hong Kong, from 1988-1998.

This study analysed monthly physico-chemical and phytoplankton data collected from 10 marine monitoring stations in Port Shelter Hong Kong, from 1988 to 1998. Cluster analysis based on all water quality parameters measured shows that the 10 monitoring stations can be grouped into three clusters: Cluster I consists of stations located in Outer Port Shelter; Cluster II consists of stations located in Inner Port Shelter and Cluster III consists of a single station in Hebe Heaven, a well-sheltered Bay in Inner Port Shelter. Factor analysis shows high positive loadings for nutrients and nutrient ratios in the first two factors of the three clusters, which indicates that the stations of the three clusters were primarily grouped according to their respective nutrient conditions. In Inner Port Shelter, and particularly Station PM5 in the sheltered Hebe Haven, the influence of surface runoff and pollution loading from the surrounding catchment area was apparent. Five stations located along a transect from Inner to Outer Port Shelter were selected for trend analysis. An increasing trend for temperature and a decreasing trend for pH are observed in all Stations except PM10 which is at the mouth of Port Shelter. The results of canonical analysis revealed that temperature and pH could also play an important role in determining the density and biovolume of the minor algal groups in Port Shelter.

Effects of the duration and timing of starvation during larval life on the metamorphosis and initial juvenile size of the polychaete Hydroides elegans (Haswell).

The larvae of the polychaete Hydroides elegans were exposed to different feeding schedules to evaluate the effects of starvation on metamorphosis and initial juvenile volume. The timing of starvation (early or late in development) and the total number of days of feeding (3, 5 or 8 days) at 24 degrees C had significant effects on the percentage of larvae that successfully completed metamorphosis and on the volume of the post-metamorphic juveniles. However, the duration of starvation (2, 4, 6 or 8 days) did not influence metamorphosis or juvenile volume. For larvae that had fed for 3 or 5 days, those that were starved at the beginning of development had a higher percentage of successful metamorphosis and produced larger juveniles compared with those starved after an initial 3-day feeding period. Larvae that had been allowed to feed for a total of 8 days were not affected by the timing of starvation. The percentage metamorphosis was lower in larvae that had fed for 3 days (16%), but was not significantly different between those that fed for 5 days (47%) compared to 8 days (47%). However, juvenile volume increased with the number of days of feeding from 0.95 nl after 3 days to 1.35 nl after 5 days, to 1.91 nl after 8 days of feeding. Discontinuous feeding reduced the proportion of larvae undergoing metamorphosis and juvenile volume compared to larvae continuously fed for only 5 days, but had no effect on larvae that fed for 8 days. The larvae of this extremely successful fouling species are remarkably tolerant of starvation and seem to require approximately 5 days of continuous feeding to achieve high levels of metamorphic success and large initial juvenile volume.

Molecular phylogeny and species identification of pufferfish of the genus Takifugu (Tetraodontiformes, Tetraodontidae).

The phylogenetic relationships and species identification of pufferfishes of the genus Takifugu were examined by use of randomly amplified polymorphic DNA (RAPD) and sequencing of the amplified partial mitochondrial 16S ribosomal RNA genes. Amplifications with 200 ten-base primers under predetermined optimal reaction conditions yielded 1962 reproducible amplified fragments ranging from 200 to 3000 bp. Genetic distances between 5 species of Takifugu and Lagocephalus spadiceus as the outgroup were calculated from the presence or absence of the amplified fragments. Approximately 572 bp of the 16S ribosomal RNA gene was amplified, using universal primers, and used to determine the genetic distance values. Topological phylogenic trees for the 5 species of Takifugu and outgroup were generated from neighbor-joining analysis based on the data set of RAPD analysis and sequences of mitochondrial 16S rDNA. The genetic distance between Takifugu rubripes and Takifugu pseudommus was almost the same as that between individuals within each species, but much smaller than that between T. rubripes, T. pseudommus, and the other species. The molecular data gathered from both analysis of mitochondria and nuclear DNA strongly indicated that T. rubripes and T. pseudommus should be regarded as the same species. A fragment of approximately 900 bp was amplified from the genome of all 26 T. pseudommus individuals examined and 4 individuals of intermediate varieties between T. rubripes and T. pseudommus. Of the 32 T. rubripes individuals, only 3 had the amplified fragment. These results suggest that this fragment may be useful in distinguishing between T. rubripes and T. pseudommus.