The influence of oyster reefs and surrounding sediments on nitrogen removal - An in-situ study along the East coast of Australia.

Oyster reefs play a crucial role in the removal of nitrogen (N) from aquatic systems by facilitating nutrient regeneration and denitrification, both in their tissues and shells and surrounding sediments. However, we still have a limited understanding about the contribution of each component of the reefs (e.g. oysters vs sediments) to N processes, and whether rates are dependent on site-specific characteristics. To address these knowledge gaps, we conducted an experiment across six oyster reefs along 1080 km of the Eastern Australian coast with different sediment characteristics. By using in-situ clear and dark incubation chambers, we assessed how benthic metabolism, nutrient and dinitrogen gas (N2) fluxes varied among the following treatments: 'oysters', 'sediments', and 'sediments + oysters' that were used to represent components of the whole reef habitat (i.e. reef matrix vs surrounding sediments vs the interaction among them, respectively), and sites. We found that during dark conditions and at siltier sites, N2 effluxes from oysters can be up to 23 times higher than sediments, while N2 effluxes from chambers with both sediments and oysters were similar to sediment treatments, and lower than oyster treatments. These results can be explained by sediment processes including nutrient assimilation by benthic microalgae and/or lower nutrient diffusion into interstitial space. Additionally, oyster treatments showed an uptake of nitrate (NO3-) that was likely converted into N2, whereas sediment treatments showed an overall release of NO3-. In dark conditions, ammonium (NH4+) fluxes remained consistent across treatments and sites, indicating that any exports from oyster excretion (in those treatments including oysters) were either counterbalanced by or comparable to exports from sediments. This study provides evidence that the crucial contribution of oyster reefs to N removal is dependent on interactions between reef components and environmental factors.

Matching Ablation Endpoints to Long-Term Outcome: The Prospective Multicenter Italian Ventricular Tachycardia Ablation Registry.

BACKGROUND: Multicenter ventricular tachycardia (VT) ablation studies have shown poorer outcomes compared with single-center experiences. This difference could be related to heterogeneous mapping and ablation strategies. OBJECTIVES: This study evaluated a homogenous simplified catheter ablation strategy for different substrates and compared the results with those of a single referral center. METHODS: This was a multicenter prospective VT ablation registry of patients with the following 4 causes of VT: previous myocardial infarction; previous myocarditis; arrhythmogenic right ventricular dysplasia; or idiopathic dilated cardiomyopathy. The procedural protocol included precise mapping and ablation steps with the combined endpoint of late potential (LP) abolition and noninducibility of VT. The long-term primary efficacy endpoint was freedom from VT. RESULTS: A total of 309 patients were enrolled. LPs were present in 70% of patients and were abolished in 83%. At the end of the procedure 74% of LPs were noninducible. The primary combined endpoint of LP abolition and noninducibility was achieved in 64% of patients with LPs at baseline. Freedom from VT at 12 months was observed in 67% of patients. In the overall study group, VT inducibility was the only predictor of freedom from VT (P = 0.013). In patients with LPs, the VT recurrence rate was lower both for patients with complete LP abolition (P = 0.040) and for patients meeting the composite endpoint (P = 0.035). CONCLUSIONS: A standardized VT mapping and ablation technique reproduced the procedural outcomes of a single referral center in a multicenter prospective study. LP abolition and noninducibility were effective in reducing VT recurrences in patients with 4 causes of cardiomyopathy. (Ventricular Tachycardia Ablation Registry; NCT03649022).

Shellfish as a bioremediation tool: A review and meta-analysis.

Over the last century, human activities have increased the amount of nutrients inputs to terrestrial and aquatic ecosystems. These activities have altered nitrogen (N) and phosphorus (P) cycling, causing substantial changes in ecosystem function such as provision of clean air and water. Strategies that reduce and remove excess nutrients are urgently needed to remediate impacted systems. Reef-forming shellfish (oysters and mussels) can play a crucial role in nutrient cycling, particularly in N removal from aquatic systems by providing substrate for microbial colonisation and enhancing microbial denitrification in the surrounding sediments. However, the potential for shellfish to enhance nutrient cycling (and denitrification) will likely vary spatially and in response to several environmental factors. Here, we used 1) a qualitative analysis to review nutrient processes occurring on shellfish; and 2) a meta-analysis to evaluate the influence of shellfish on benthic metabolism and nutrient cycling in surrounding sediments, and how that is influenced by environmental factors such as grain size, seasonality, water body type, and tidal position. Overall, we found that shellfish increased oxygen consumption, with consequent release of ammonia (NH4+) and phosphate (PO43-) from shellfish and their surrounding sediments. These parameters did not depend on grain size, water body type and tidal height, but the release of PO43- was variable between seasons, being highest during summer and autumn. Shellfish presence also enhanced denitrification measured as dinitrogen gas (N2) efflux on both reefs and sediments. Denitrification was highest in lagoons; in sandy sediments; and during the warmest season (summer). Thus, our findings highlight that environmental context can mediate the effects of shellfish reefs on sediment function. This information is important for managers seeking to use these animals as an effective bioremediation tool.

Variations in benthic fluxes of sediments near pier pilings and natural rocky reefs.

Marine artificial structures such as pilings are replacing natural habitats, and modifying surrounding areas, often resulting in local decreases in species diversity and facilitation of bioinvasion. Most research on the impacts of artificial structures in marine ecosystems has primarily focused on rocky bottom habitats and biodiversity, overlooking the effects of these structures on the functioning of nearby sedimentary habitats. Here we compared, for the first time, benthic metabolism (O2 fluxes) and sediment-water nutrient (inorganic nitrogen, phosphate, and dissolved organic nitrogen) fluxes in shallow water sediments adjacent to pilings and natural reefs. We also measured sediment properties (grain size, total organic carbon, total nitrogen, C:N ratio and chlorophyll-a content). We found that sediments near pilings were generally finer with greater C:N ratios than those near reefs, while differences in other sediment properties between types of habitats were dependent on the site. We found significant differences in the oxygen consumption, primary productivity, and net ecosystem metabolism in sediments around pilings compared to sediments near natural reefs, but these patterns differed by site. Net nutrient fluxes were similar in sediments near pilings and reefs at both sites. This study showed that although pilings can be associated with changes in the functioning of sedimentary habitats, patterns and the direction of change seem to vary depending on local conditions.

Season specific influence of projected ocean changes on the response to cadmium of stress-related genes in Mytilus galloprovincialis.

Anthropogenic inputs of carbon dioxide in the atmosphere are driving ocean warming and acidification. The potential threat represented by these changes for marine species could be amplified in coastal areas, characterized by higher levels of pollutants. In addition, temperate organisms may exhibit a different seasonal tolerance to stressors influenced by fluctuations of environmental and physiological factors. In this study, Mediterranean mussels Mytilus galloprovincialis collected both in summer and winter were exposed to combinations of two temperatures (SST, seasonal surface temperature and SST+5 °C) and two levels of pH (8.20 and 7.40) in clean or cadmium contaminated seawater (20 µg/L Cd). mRNA levels of genes related to metal-induced stress response were investigated, including metallothionein mt-20, heat-shock protein hsp70, superoxide dismutase Cu/Zn-sod, catalase cat, glutathione peroxidase gpx1 and glutathione S-transferase gst-pi. To further elucidate if tissues with different physiological roles could exhibit different responsiveness, such analyses were carried out in digestive gland and in gills of exposed mussels. mt-20 mRNA increase after Cd-exposure was higher in the digestive gland than in the gills, with few modulations by temperature or pH only in the latter. Acidification, alone or in combination with other stressors, increased hsp70 mRNA, with seasonal- and tissue-specificities (higher in summer and in digestive gland). Among antioxidants, gpx1 mRNA was affected by Cd in both tissues and seasons, with further modulations due to pH and temperature variation tissue- and season-specific; in winter the combination of Cd, warming and acidification affected Cu/Zn-sod both in digestive gland and gills and cat only in gills, while weak seasonal variations were observed for gst-pi transcripts only in digestive gland. The overall results highlighted the importance of considering seasonality and responsiveness of different tissues to predict the effects of sudden changes in environmental parameters on responsiveness to and toxicity of chemicals in marine coastal organisms.

Sediment bacterial communities associated with environmental factors in Intermittently Closed and Open Lakes and Lagoons (ICOLLs).

Intermittently Closed and Open Lakes and Lagoons (ICOLLS) are important coastal systems that are periodically separated from the ocean by a sand barrier or a berm. In urban ICOLLs, continuous inputs of organic material and nutrients into coastal lagoons are contributing to eutrophic conditions that, together with natural environmental factors have implications for the resident sediment bacterial communities. We used molecular tools to investigate the ecological communities of four ICOLLs; Narrabeen, Dee Why, Curl Curl and Manly in Sydney, Australia, which have been subjected to increasing pressure from anthropogenic activities over the last century. We used targeted gene sequencing of the prokaryotic 16S ribosomal RNA gene to describe the bacterial diversity and community structure and discuss differences with respect to environmental factors at the ICOLL scale (e.g. size, shape, normalised N loading) and site scale (e.g. water and sediment quality) within each lagoon. Due to differences in hydrological patterns, we expected that sediment bacterial communities would be more heterogenous in displacement-dominated lagoons (Curl Curl and Manly) than the mixing-dominated lagoons (Narrabeen and Dee Why). Interestingly, we did not find strong relationships between the main bacterial groups and distance from the lagoon entrance (used as a proxy for salinity and silt) in the displacement-dominated lagoons. Moreover, we found that α diversity was highest in Narrabeen and Manly lagoons despite differences in lagoon size and shape. Furthermore, while bacterial community structure was similar in Curl Curl and Dee Why, communities in Manly and Narrabeen differed along temperature/TOC and salinity/silt gradients respectively. In Manly lagoon, we found relatively more anaerobic bacteria such as Epsilonbactereota which is involved in the oxidation and reduction of sulfur compounds. Moreover, we identified several bacterial taxa (including sulfur metabolising Chlorobiaceae) related to increasing TOC that could be investigated further as potential indicators of excess enrichment.

Two additional reports of deletion on the short arm of the Y chromosome.

Deletions on the short arm of the Y chromosome involving the amelogenin Y gene (AMELY), located on Yp11.2, can be misleading for sex typing with serious consequences in forensic applications and prenatal diagnosis. In this study, we describe two AMELY null cases concerning two unrelated Italian males from Northeast Italy. PCR amplification of short tandem repeats on the Y chromosome (Y-STRs) showed a lack of AMELY and DYS458 markers. The presence of all the other markers located on the Y chromosome and of the SRY gene in both samples led us to conclude that a deletion had occurred in a portion of the short arm of the Y chromosome. Twenty-three Y-specific sequence tagged sites (STSs) were chosen to delineate the deletion's length, which was estimated to be in the range of 3.35-3.87Mb for one sample and 1.51-2.58Mb for the other. These and previous findings suggest that in all cases where potential AMELY drop out has occurred, it should be used additional specific Y chromosome markers or human DNA quantification methods that specifically quantify male DNA using target male genomic markers, which not being located within the deletion regions, allow an accurate sex identification.

STR typing of archival Bouin's fluid-fixed paraffin-embedded tissue using new sensitive redesigned primers for three STR loci (CSF1P0, D8S1179 and D13S317).

Three new mini-STR primer sets are suggested for three conventional STRs, CSF1P0, D8S1179 and D13S317, included in multiplex PCR kits commercially available and commonly used for DNA typing in forensic applications. The primer pairs for the three loci were redesigned in order to reduce or eliminate the flanking regions of the polymorphism obtaining amplification products, which have dimensions less than 120bp in size. A comparison of results for typing carried out with the newly designed primers on DNA extracted from 100 blood samples provided by healthy donors, previously typed with conventional STRs, showed no genotype difference underlining their precision and reproducibility. The forensic usefulness of the new mini-STR primers was evaluated on highly degraded DNA from casework samples (e.g. archival post-mortem Bouin's fluid-fixed paraffin-embedded tissue specimens) for which commercial STR kit had proven inefficient.

Development and forensic validation of a new multiplex PCR assay with 12 X-chromosomal short tandem repeats.

One multiplex system for the co-amplification of 12 X-chromosomal short tandem repeats (STRs) DXS7132, DXS8378, DXS6809, DXS7133, DXS6789, DXS7424, GATA172D05, HPRTB, DXS7423, GATA31E08, DXS101, DXS6807 and amelogenin was analysed in a sample of 200 (100 males and 100 females) unrelated healthy individuals living in Northern Italy. The chi2-test for genotype distribution of the X-chromosomal STRs showed no significant deviation from the Hardy-Weinberg equilibrium (HWE). Allele frequencies between female and male samples were not significantly different in all examined markers. In the kinship cases involving 40 family trios with daughter and 10 father/daughter duos, no mutation was detected. The combined power of discrimination (PDc) of the 12 X-STRs for both females and males was PDc > 0.999999.

Intracoronary ST-segment shift soon after elective percutaneous coronary intervention accurately predicts periprocedural myocardial injury.

BACKGROUND: Elevation of cardiac biomarkers after coronary angioplasty (percutaneous coronary intervention [PCI]) reflects periprocedural myocardial damage and is associated with adverse cardiac events. We assessed whether periprocedural myocardial damage that occurs despite successful PCI could be rapidly and easily identified by intracoronary ST-segment recording with the use of a catheter guidewire. METHODS AND RESULTS: In 108 consecutive stable patients undergoing elective single-vessel PCI, we recorded unipolar ECG from the intracoronary guidewire in the distal coronary before PCI and 2 minutes after the last balloon inflation. After PCI, intracoronary ST-segment shift > or = 1 mm from baseline was considered significant. Troponin I levels were measured at baseline and at 8 and 24 hours after intervention, and myocardial damage was defined as troponin I increase above the upper normal value after intervention. All patients had normal cardiac marker values before PCI, and PCI was successful in all (residual stenosis < 20%, Thrombolysis in Myocardial Infarction grade 3 flow). After PCI, long-term follow-up data were collected; myocardial damage was detected in 50 patients (46%), although abnormal creatine kinase-MB values were documented in only 11 (10%). Significant intracoronary ST-segment shift after PCI was present in 40 patients (37%; group A) and absent in the remaining 68 (63%; group B). Procedural myocardial damage was documented in 37 group A patients (93%) and in 13 group B patients (19%; P<0.001); significant ECG changes were found on standard ECG after intervention in only 5 patients (13%) and 1 patient (1%) (P<0.05). Sensitivity of intracoronary ST-segment shift for predicting myocardial damage was 74%, and specificity was 95%, with positive and negative predictive values of 93% and 81%, respectively. On multivariate analysis, intracoronary ST-segment shift was the sole independent predictor of myocardial damage (odds ratio, 54.1; 95% confidence interval, 12.1 to 240; P<0.0001). At a median follow-up of 12+/-5 months, major coronary event-free survival was significantly worse in group A patients (log-rank test chi2=4.0; P<0.05). CONCLUSIONS: After successful single-vessel PCI, intracoronary ST-segment shift allows the prompt and inexpensive identification of patients developing myocardial injury, who may require adjunctive therapy and longer in-hospital stay.

Intracoronary ST segment evolution during primary coronary stenting predicts infarct zone recovery.

In patients with acute myocardial infarction (AMI), early ST segment elevation resolution on ECG predicts myocardial reperfusion and LV recovery. Intracoronary ECG is more sensitive than surface ECG to detect regional ischemia. In patients undergoing primary percutaneous coronary intervention (PCI), we investigated if failed myocardial reperfusion, despite successful infarct vessel recanalization, could be rapidly and easily identified by intracoronary ST segment monitoring from guidewire recording. We recorded intracoronary and standard ECG during primary coronary stenting (PCI) in 50 patients with AMI (59 +/- 11 years; anterior AMI in 66%). All patients had a successful PCI and underwent 2D echocardiography soon after PCI and 6 months later. Following PCI, intracoronary ST resolution >/= 50% from baseline was documented in 39 patients (78%; group A; from 11 +/- 8 to 1 +/- 2 mm) but not in 11 (22%; group B; from 11 +/- 8 to 8 +/- 5 mm). Group A had slightly shorter ischemic time (202 +/- 94 vs. 238 +/- 112 min in B; P = 0.2) and smaller peak CK values (2,752 +/- 2,038 vs. 4,802 +/- 3,671 U/L in B; P = 0.02). After PCI, ST resolution was found on standard ECG in 34 (87%) group A and in 3 (27%) group B patients. At 6-month follow-up, left ventricular ejection fraction was greater in group A (47% +/- 8% vs. 39% +/- 8% in B; P < 0.001) with improved wall motion score index (from 2.2 +/- 0.3 to 1.7 +/- 0.3 in A; from 2.3 +/- 0.4 to 2.1 +/- 0.4 in B; P < 0.001). There were no significant differences between intracoronary and standard ECG for sensitivity (92% vs. 86%) and specificity (62% vs. 57%) to predict improved infarct zone recovery after 6 months. ST elevation resolution on intracoronary recording during PCI predicts infarct zone recovery. Monitoring ST segment evolution by intracoronary ECG allows prompt and inexpensive identification in the catheterization laboratory of those patients without myocardial reperfusion, who may require adjunctive therapeutic interventions after successful infarct vessel recanalization.