Denitrification plays a central role in the global nitrogen cycle, reducing and removing nitrogen from marine and terrestrial ecosystems. The flux of nitrogen species through this pathway has a widespread impact, affecting ecological carrying capacity, agriculture, and climate. Nitrite reductase (Nir) and nitric oxide reductase (NOR) are the two central enzymes in this pathway. Here we present a previously unreported Nir domain architecture in members of Phylum Chloroflexi. Phylogenetic analyses of protein domains within Nir indicate that an ancestral horizontal transfer and fusion event produced this chimeric domain architecture. We also identify an expanded genomic diversity of a rarely reported nitric oxide reductase subtype, eNOR. Together, these results suggest a greater diversity of denitrification enzyme arrangements exist than have been previously reported.

The relatively long turnaround time and low sensitivity of traditional blood culture may delay the effective antibiotic therapy in patients with bloodstream infection (BSI). To reduce the morbidity and mortality of BSI, a rapid and sensitive pathogen detection method is urgently required. Acinetobacter baumannii and Klebsiella pneumonia are two major microorganisms responsible for BSI. Here we reported a novel droplet digital PCR (ddPCR) method that can detect A. baumannii and K. pneumonia in whole blood samples within 4 h, with a specificity of 100% for each strain and limit of detection at 0.93 copies/microliter for A. baumannii and 0.27 copies/microliter for K. pneumonia. Clinical validation in 170 patients with suspected BSIs showed that, compared with blood culture that reported 4 (2.4%) A. baumannii cases and 7 (4.1%) K. pneumonia cases, ddPCR detected 23 (13.5%) A. baumannii cases, 26 (15.3%) K. pneumonia cases, and 4 (2.4%) dual infection cases, including the 11 positive patients reported by blood culture. In addition, the positive patients reported by ddPCR alone (n = 42) had significantly lower serum concentrations of procalcitonin and lactate, SOFA and APACHE II scores, and 28-day mortality than those reported by both blood culture and ddPCR (n = 11), suggesting that patients with less severe manifestations can potentially benefit from the guidance of ddPCR results. In conclusion, our study suggests that ddPCR represents a sensitive and rapid method to identify causal pathogens in blood samples and to guide the treatment decisions in the early stage of BSI.

Introduction Pregnancy is a process that causes several physiological changes including all systems as well as cardiovascular system. Ventricular hypertrophy and dilation of cardiac chambers are seen as a result of these changes. Although there are studies about pregnancy-related changes in echocardiographic examination; there is no data about the long-term effects of parity on these alterations. Therefore, we have evaluated the long-term effect of pregnancy on right ventricular (RV) dilation and RV hypertrophy and their relation to the parity number. Methods This prospective study included a total of 600 women (200 consecutive women who had no parity, 200 women who had a parity number of 1 to 4 and 200 women who had a parity number of more than 4). Right chambers’ measurements were compared between the groups. Results In echocardiographic analysis, RV and right atrial dimensions and areas and RV wall thickness were higher in parous women. On the other hand, RV systolic function parameters were significantly lower in parous women. These significant changes showed a gradual increase or decrease by increasing parity number. There were also independent relationship between the number of parity and RV hypertrophy even after adjustment for several confounders. Conclusion Pregnancy-related physiological changes mostly resolve after delivery. This study about long-term effects of pregnancy on RV has demonstrated that there is a significant relation between the number of parity and either RV dilation or RV hypertrophy. Each parity had also additive effect on these changes.

Kawasaki disease (KD) is a self-limited vasculitis with significant morbidity and even mortality if not treated early. The diagnosis and timely treatment in children younger than 3 months is challenging, most of them have an incomplete or atypical presentation. Coronary artery abnormalities are frequent in this type of patients. We present a 6-week-old female infant with Kawasaki disease who developed a giant coronary aneurysm. The timely diagnosis and promptly treatment as well as the echocardiographic and multimodality follow-up allowed us to improve our clinical approach and management.

Abnormal pulmonary venous flow patterns on fetal echocardiography and a nutmeg lung pattern on fetal magnetic resonance imaging are seen in patients with pulmonary venous stenosis. The association between these findings and the degree of pulmonary venous stenosis remains unknown. We report an extremely rare case of a fetus diagnosed with hypoplastic left heart syndrome complicated by an absent atrial septum and supracardiac total anomalous pulmonary venous connection with left pulmonary venous congestion. This case suggests that compared to non-pulsatile continuous pulmonary venous flow, the nutmeg lung pattern can only be observed with severe pulmonary congestion and advanced pulmonary lymphangiectasia.

Objective: To estimate the association between maternal origin and obstetric anal sphincter injury (OASI), and assess if associations differed by length of residence. Design: Population-based cohort study. Setting: The Medical Birth Registry of Norway. Population: Primiparous women with vaginal livebirth of a singleton cephalic fetus between 2008 and 2017 (n=188 658). Methods: Multivariable logistic regression models estimated aORs for OASI with 95% CI by maternal region of origin and birthplace. We stratified models on length of residence and paternal birthplace. Main outcome measures: OASI. Results: Overall 6 373 cases of OASI were identified (3.4% of total cohort). Women from South Asia were most likely to experience OASI (6.2%; aOR 2.24, 95% CI 1.93–2.60), followed by those from Southeast/East Asian/Pacific (5.7%; 1.83, 1.64–2.04), and Sub-Saharan Africa (5.2%; 1.97, 1.72–2.26), compared to women originating from Norway. Among women born in the same region, those with short length of residence in Norway (0–4 years), showed higher odds of OASI. Migrant women across most regions of origin had reduced risk of OASI if they had a Norwegian compared to foreign-born partner. Conclusions: Primiparous women from Asian regions and Sub-Saharan Africa had up to two-fold risk of OASI, compared to women originating from Norway. Migrants with short residence and those with a foreign-born partner had higher risk of OASI, implying that some of the risk differential is due to sociocultural factors. Some migrants, especially new arrivals, may benefit from special attention during labour to reduce morbidity and achieve equitable outcomes.

To investigate the frame of reference with the downward displacement of the posterior leaflet and anterior leaflet of tricuspid valve in children by ultrasound.The downward degree of anterior and posterior tricuspid valve was evaluated with tricuspid annulus and coronary sinus as reference structures under ultrasound, and the position of tricuspid regurgitation orifice was shown by color ultrasound. Color Doppler flow imaging showed that the position of tricuspid regurgitation orifice moved down obviously in all 42 children. One case showed 2.2cm from the root of the anterior valve to the tricuspid annulus in the two-chamber and four-chamber view of the apical right heart. Color Doppler can show that the position and direction of tricuspid regurgitation orifice are obviously deviated to the anterolateral side. The obvious deviation of the tricuspid regurgitation orifice to the anterolateral direction may be an ultrasonic sign for diagnosing the downward displacement of the anterior tricuspid valve in children.

We report first in man implantations of the newly designed Evita-open-NEO hybrid prosthesis (HP) for complex aortic arch disease from 3 different countries in Asia-Pacific including suggestions on how to proceed with optimized perioperative coagulation.

A document by Karunesh Singh . Click on the document to view its contents.

Some iconic symbols representing numbers are more animated, picturesque, and impressive. Here we describe a kind of new numeral symbols from one to septillion. These symbols are regular and simple. Meanwhile it is easy to write and remember these number symbols. They are composed of 27 basic components. After these basic components are regularly combined, simple and effective numeral symbols from one to septillion can be generated. They might be used in games, movies, animations, computer science and even in other arts.

We present a patient with congenital Glanzmann Thrombasthenia, who developed recurrent venous thrombosis. Over time, she developed the clinical picture of a myeloproliferative neoplasm, being JAK2 positive. This case clearly indicates that the platelet aIIbβIII integrin (lacking in Glanzmann thrombasthenia) does not have a role in thrombosis in MPN

Neuropathology studies of amyotrophic lateral sclerosis (ALS) and animal models of ALS reveal a strong association between aberrant protein accumulation and motor neuron damage and activated microglia and astrocytes, the resident CNS innate immune cells. While the role of neuroinflammation in the pathology of ALS is unclear imaging studies support the idea that innate immune activation occurs early disease in both humans and rodent models of ALS. In addition to innate immunity, emerging studies also reveal the presence of peripheral monocytes, macrophages, and lymphocytes in the CNS as well as at the neuromuscular junction. To better understand the association of neuroinflammation (innate and adaptive) with disease progression paraclinical studies including the use of biomarkers and imaging modalities allow monitoring of immune parameters in the disease process. Such approaches are important for patient stratification, selection, and inclusion in clinical trials, as well as to provide readouts of response to therapy. Here, we discuss the different imaging modalities e.g., MRI, MRS, PET as well as other approaches including biomarkers of inflammation in ALS, aid the understanding of the underlying immune mechanisms associated with motor neuron degeneration in ALS.

A Preliminary Argument for the Selective Use of the Robicsek WeaveJohn S. Ikonomidis MD, PhDDivision of Cardiothoracic Surgery, University of North Carolina at Chapel HillWord Count: 886References: 4Address correspondence to:John S. Ikonomidis MD, PhDProfessor and Chief,Division of Cardiothoracic SurgeryUniversity of North Carolina at Chapel Hill3034 Burnett Womack Building160 Dental Circle,Chapel Hill, NC27599e-mail: john_ikonomidis@med.unc.eduTel: (919) 966-3381Proper execution of median sternotomy and its subsequent closure are critical to the success of cardiac surgical outcomes. It is essential that the sternum be divided directly in the midline, and table fractures must be avoided if at all possible by avoiding excessive spreading if the sternum for exposure of the heart. Multiple methods have been described regarding primary sternal closure technique, but the conventional technique of wire circlage, either linear or figure-of-eight, has endured and is also the most cost-effective. Sternal wound complications have an incidence of 0.8% to 1.5% patients, and this number rises to as high as 8% when bilateral internal mammary artery harvest is undertaken. Further established risk factors for deep sternal wound complications include breaches in sterility in the operating room, lengthy operations, re-exploration for bleeding, undrained retrosternal hematoma, incomplete wound closure, obesity, advanced age, diabetes, chronic obstructive pulmonary disease, hospital acquired pneumonias, renal failure, requirement for dialysis, and prolonged mechanical ventilation. Mortality from sternal dehiscence and related complications ranges from 6% to 70%. It is generally felt that early treatment reduces mortality.1Deep sternal wound complications and dehiscence were once thought to be highly feared and challenging complications of cardiac surgery. Modern primary closure techniques, tissue flap coverage options, and negative pressure wound therapy have made these complications more manageable. Nevertheless, it behooves surgeons to avoid this complication due to its considerable negative clinical impact.There are many methods currently available for reconstruction of the sternum after its dehiscence, the most common of which is the sternal weave first described by Robicsek and colleagues in 1977.2 This technique is often used to reinforce the sternum with primary sternal closure in instances where the sternotomy was off the midline leaving a thin weak section of sternum on one side or where some fracturing has occurred, but has also been used as a first line for sternal reconstruction after its dehiscence from primary closure. Data are not available regarding the overall success rate of reinforcement using the Robicsek weave, but at least one multicenter, randomized controlled trial showed that in patients with an increased risk for sternal instability and wound infection after cardiac surgery, sternal reinforcement using the Robicsek technique prior to primary sternal closure did not reduce dehiscence rate.3In addition to the above, antecedent sternal weaving weave may complicate further attempts at sternal closure should dehiscence recur. In this issue of the Journal of Cardiac Surgery,4Seyrek et al. conducted a retrospective review of patients at a single institution with noninfectious sternal dehiscence (NISD) after median sternotomy who received thermoreactive nitinol clips (TRNC) for sternal closure. The authors studied 34 cases who received TRNC treatment between December 2009 and January 2020 out of 283 patients with NISD who underwent sternal refixation. These cases were divided into two groups: patients who had a previously failed Robicsek procedure before TRNC treatment (group A, n=11) and patients who had been directly referred to TRCN treatment (group B, n= 23). The results showed that the postoperative complication rate and length of hospital stay was significantly higher with use of the Robicsek weave. Further, operative time was significantly shorter and blood loss was significantly lower in patients referred for sternal refixation without having first undergone a Robicsek weave.Part of the reason for the above results may lie with the surgical requirements for performance of the Robicsek weave. Substernal and lateral dissection is required to define the margins of the sternum before placing the weave. This increases the technical difficulty of the reclosure operation and puts the patient at risk for inadvertent injury to the heart, great vessels, and other mediastinal structures. This dissection may also compromise blood flow to the sternal half. Further, intercostal arteries may be squeezed by weave as it runs anteriorly and posteriorly around the ribs, which may occlude blood supply to the sternum. This could worsen pre-existing ischemia, which would delay sternal healing, promote bacterial colonization, and cause bone necrosis and additional sternal fragmentation, thus complicating any additional closure attempts.Use of TRNC may represent an advance in sternal reconstruction therapy due to the simplicity of use and lack of requirement for a complex mediastinal dissection prior to application. The authors contend that a previously failed Robicsek procedure caused significantly higher morbidity, additional operative risk and lower success rate in later TRNC treatment of high-risk cases and hence speculate that patients at high risk for sternal separation should proceed directly to TRNC treatment. In the light of the above study, this approach seems reasonable, but a prospective trial should be considered to provide the definitive answer.

Objective: To compare plasma free amino acid (FAA) and carnitine levels in pregnant women with and without hydatidiform mole (HM). Methods: This prospective study was conducted in patients admitted to Harran University Medical Faculty Hospital Obstetrics Clinic between January and November 2018. Twenty-three pregnant women with HM, and 24 healthy pregnant women as controls were enrolled in the study. The venous blood samples were collected, and, FAA and carnitine concentrations were measured in plasma using liquid chromatography/tandem mass spectrometry (LC-MS/MS). Results: The levels of alanine, arginine, and valine from the 14 amino acids examined were significantly lower in the HM group than in the healthy group (475.46 ± 103.55 µmol/L vs 556.99 ± 105.16 µmol/L, p = 0.019; 428.87 ± 138.91 µmol/L vs 547.11 ± 140.20 µmol/L, p = 0.009; and 290.96 ± 97.10 µmol/L vs 358.61 ± 75.40 µmol/L, p = 0.03; respectively). In addition, several carnitines, C8DC, C16:1, and C18, of the 27 carnitines examined were significantly higher in the HM group than in the control group (0.07 ± 0.06 µmol/L vs 0.03 ± 0.01 µmol/L, p = 0.021; 0.18 ± 0.09 µmol/L vs 0.10 ± 0.06 µmol/L, p = 0.03; and 0.07 ± 0.02 µmol/L vs 0.03 ± 0.01 µmol/L, p = 0.021; respectively). Conclusion: This study demonstrated that a decrease in some plasma FAAs and an increase in some plasma carnitine levels might be effective in the pathogenesis of HM.

Environmental DNA for biomonitoringJan Pawlowski1,2,3, Aurélie Bonin4, Frédéric Boyer5, Tristan Cordier1,6, Pierre Taberlet5,71 Department of Genetics and Evolution, University of Geneva, Geneva, Switzerland2 Institute of Oceanology, Polish Academy of Sciences, Sopot, Poland3 ID-Gene Ecodiagnostics, Geneva, Switzerland4 Department of Environmental Science and Policy, Università degli Studi di Milano, Milano, Italy5 Laboratoire d’Ecologie Alpine (LECA), CNRS, Université Grenoble Alpes, Grenoble, France6 NORCE Climate, NORCE Norwegian Research Centre AS, Bjerknes Centre for Climate Research, Jahnebakken 5, 5007 Bergen, Norway7 Tromsø Museum, UiT – The Arctic University of Norway, Tromsø, NorwayCorresponding authorJan.Pawlowski@unige.chIn 2012, Molecular Ecology published a special issue on environmental DNA, which provided an overview of the field of eDNA research and presented a selection of papers on eDNA studies (Taberlet, Coissac, Hajibabaei, & Rieseberg, 2012). This special issue also introduced the concept of Biomonitoring 2.0, advocating for the use of DNA-based identification of taxa in biodiversity surveys and ecosystem assessment (Baird & Hajibabaei, 2012). Since then, hundreds of papers have been published covering various aspects of eDNA-based biomonitoring from single-species detection to community studies and environmental impact assessments. Numerous reviews have summarized these studies for both freshwater and marine environments (e.g. Bohmann et al., 2014; Thomsen & Willerslev, 2015).The progress made in the eDNA field during these last ten years has been spectacular (Taberlet, Bonin, Zinger, & Coissac, 2018). Although the basic concepts and workflow of DNA barcoding and metabarcoding have not changed, the technological advances in high-throughput sequencing have greatly facilitated the access to eDNA data. It has become possible to monitor biodiversity with unprecedented precision and depth. Massive environmental genomic datasets have been rapidly generated at relatively low cost. The analysis of these datasets using machine learning and other taxonomy-free approaches opened wide the doors for using new groups of bioindicators to infer ecological status (Cordier et al., 2018; Cordier, Lanzén, Apothéloz-Perret-Gentil, Stoeck, & Pawlowski, 2019; Pawlowski et al., 2018). At the same time, constant efforts to fill gaps in barcoding reference databases considerably increased the effectiveness of taxonomic identification of eDNA data (Weigand et al., 2019).Astonishingly, these rapid advances in eDNA-based technologies are rather timidly implemented in routine biomonitoring (Hering et al., 2018; Shackleton et al., 2021). Although the concept of Biomonitoring 2.0 is widely endorsed, its acceptance in practice is hampered for various reasons. There is no consensus whether eDNA-based biomonitoring should only apply to conventional bioindicators (Renovate) or should also include new bioindicators (Rebuild) or new taxonomy-free approaches (Revolutionize) (see Fig. 1). Moreover, three main steps on the roadmap from eDNA to biomonitoring are not developed equally. The main attention is given to the development and optimization of eDNA data generation and analysis. The standardization of eDNA methods and their translation into legislatory framework remain at a very early stage. One of the main issues impeding the application of eDNA-based tools concerns the lack of congruence between the results of traditional and molecular analyses (Aylagas et al., 2020). It is expected that the new method is “safe to use” only if it provides the same or almost same results as the conventional one. However, obtaining such perfect congruence is often impossible because the character of data is very different (e.g., abundance of individuals vs abundance of eDNA reads). Moreover, the eDNA “ecology” can hardly be translated directly into species ecology. There are also numerous biological and technical biases that can affect the generation and processing of eDNA data, impacting their interpretation.This special issue addresses some of these challenges by presenting the latest advances in eDNA field and discussing their strengths and limitations when applied to routine biomonitoring. The issue comprises 29 papers grouped into four sections and covering different aspects of eDNA applications. It is accompanied by an opinion paper, which clarifies the eDNA terminology in relation to its use in biomonitoring (Pawlowski, Apothéloz-Perret-Gentil, & Altermatt, 2020). The first section comprises a series of studies using new analytical tools (e.g. machine learning), new types of bioindicators and genomic data (e.g. shotgun sequencing) for the assessment of ecological status. It is followed by a section dedicated to fish eDNA, whose application in biomonitoring is the most advanced. The third section comprises papers dealing with various methodological aspects and the comparison between conventional and molecular methods. The final section presents few examples of eDNA applications for biodiversity surveys and population genetics.Novel approaches to monitor ecosystemsThe development of environmental genomics enables monitoring of microbial and meiofaunal communities that were previously inaccessible when using conventional methods. However, our knowledge of the ecology of these communities is very limited and therefore new analytic approaches are necessary to integrate them into routine bioassessment. This section begins with a review of implementation strategies for the application of environmental genomics in ecological diagnostics (Cordier et al., 2021). The authors introduce four broad categories of possible strategies, including (1) DNA-based taxonomic identification of known bioindicators, (2) taxonomy-free discovery of new bioindicators, (3) structural community metrics, and (4) functional community metrics. Each of these strategies is adapted to a particular type of data (metabarcoding, metagenomics, metatranscriptomics) and rely on different computational analyses in order to provide an assessment of the ecological status.Among the different analytical tools, machine learning seems to be the most promising way to predict the ecological status (Cordier et al., 2018, 2019). In this issue, its performance is tested in the case of the benthic diatoms index widely used in the assessment of ecological quality of rivers and streams (Apothéloz-Perret-Gentil et al., 2021). This study shows that supervised machine learning performs better than the taxonomic assignment, but its predictions are similar to those obtained using a taxonomy-free molecular assignment approach. Moreover, the efficiency of a taxonomic assignment method strongly depends on the completeness of the reference database, highlighting the need to fill in the existing gaps, particularly in the case of bioindicator taxa.The ability of de novo prokaryotic bioindicators to predict multiple anthropogenic impacts on estuarine and coastal benthic communities is demonstrated by Lanzén, Mendibil, Borja, & Alonso-Sáez (2021). The authors compare their results to the traditional macrofauna-based indices and discuss various advantages of using microbial bioindicators as they are more sensitive to different abiotic pressures. Similar conclusions were reached in the case of environmental impact assessment of marine aquaculture (Frühe et al., 2021) and the oil and gas industry (Mauffrey et al., 2021). Both studies demonstrate the effectiveness of machine learning andde novo microbial bioindicators and promote their use for benthic monitoring in marine environments.The last two papers in this series explore new directions for the further development of ecogenomic diagnostics. Broman et al. (2021) use environmental RNA (eRNA) shotgun sequencing to analyse the impact of organic enrichment on benthic micro-eukaryotic communities. Compared to eDNA metabarcoding that is used in the majority of studies, eRNA shotgun data has the advantage to overcome the potential biases of PCR amplification and to better capture the organismic response to environmental pressures by targeting predominantly active cells. Ibrahim et al. (2021) use historical eDNA metabarcoding data to analyze the impact of eutrophication on lake phytoplankton in the 20thcentury. This study demonstrates the potential of paleo-metabarcoding to characterize past biodiversity and establish reference conditions for future monitoring.Refining fish eDNA surveysThe second series of papers concerns the use of eDNA to monitor fish diversity. We focus on fish because they are among the most important groups of bioindicators and also because their study from an eDNA perspective is the most advanced (Pont et al., 2021). The barcoding reference database of common fish species in some regions is close to completeness (Knebelsberger, Dunz, Neumann, & Geiger, 2015), fish-specific markers are well defined (M. Miya et al., 2015; Valentini et al., 2016; Zhang, Zhao, & Yao, 2020) and protocols for fish eDNA sampling and processing are well established (Masaki Miya, Gotoh, & Sado, 2020; Valentini et al., 2016). Currently, considerable efforts are directed to solve the most challenging issue, which is related to quantitative fish eDNA data and its application for inferring fish indices in routine biomonitoring.Two papers address this issue by proposing different approaches to estimate fish abundance from eDNA data. Fukaya et al. (2021) use numerical hydrodynamic models to simulate the spatial and temporal distribution of fish eDNA in aquatic environments. By integrating the models to the measures of eDNA concentration, the authors obtained estimates of fish population abundance comparable to those obtained by the quantitative echo sounder method. Yates et al. (2021) improve the correlation between eDNA concentration and fish abundance by integrating allometric scaling coefficients. Such coefficients can help adjust the values of eDNA production taking in consideration density, biomass and metabolic rates characteristic to a given taxon.A better understanding of the “ecology” of fish eDNA, and particularly how its temporal and spatial distribution is shaped by abiotic and biotic factors, is the subject of the following papers. Littlefair et al. (2021) tested how seasonal variations in thermal stratification influence the distribution of fish eDNA in lakes. The authors show that eDNA distribution follows lake stratification and the thermal niche of the species, which in turn may affect its detection in certain seasons. The distribution of fish and amphibian eDNA in a lentic system was investigated experimentally by Brys et al. (2021). This study indicates high eDNA decay rates and limited dispersal, reinforcing the accuracy of eDNA-based monitoring for retrieving the spatiotemporal occupancy patterns. The advantages of using eDNA for survey of fish populations were also demonstrated by other papers in this section. McColl-Gausden et al. (2021) showed that eDNA metabarcoding is generally more sensitive than electrofishing for conducting fish surveys in freshwater streams, while Aglieri et al. (2021) demonstrate strong complementarity of eDNA-based analysis with visual and capture-based methods in the survey of coastal fish communities.Methodology and comparison with conventional methodsGeneral acceptance of molecular methods in biomonitoring requires their benchmarking against conventional morphotaxonomy-based approaches. This is commonly achieved by processing the same samples in parallel using different methods and by assessing how the molecular data fit to the results of traditional approaches, considered as a ground truth. The papers of this section compare the results of eDNA metabarcoding vs bulk DNA metabarcoding vs different morphology-based approaches. They also present and discuss the biases of molecular methods and propose solutions to improve the outcomes of molecular data generation and processing.The section begins with the three comparative studies of marine biomonitoring. Suter et al. (2021) evaluate the performance of water eDNA and bulk DNA metabarcoding in assessing the biodiversity of zooplankton in open ocean, currently monitored by using continuous plankton recorders. The study shows that both methods recover more species than morphological analyses, however, their efficiency depends on the sampling method and selected marker. They conclude that eDNA metabarcoding is very promising, but it still requires some refinement and standardization before it can be routinely used for zooplankton biomonitoring. Similar conclusions are drawn from the comparison of sediment DNA metabarcoding and macrofauna surveys applied to monitor benthic impacts of salmon farms (He et al., 2021). Although the authors found a certain coherence in relative abundance of common macrofauna bioindicators inferred from morphological and eDNA data, they observed that the correlation with organic enrichment was much stronger for meiofauna, which is not usually included in biomonitoring studies. Significant differences were also found between water eDNA samples and bulk DNA extracts from adjacent benthic communities (Antich et al., 2021). The authors concluded that water eDNA is a poor proxy for the analysis of benthic communities, although they do not exclude that the use of taxon-specific markers could improve the congruence between eDNA and bulk DNA metabarcoding data.The importance of marker selection has also been emphasized in the case of freshwater macrobenthos metabarcoding. The performance of different markers, with focus on key insect orders (Ephemeroptera, Plecoptera and Trichoptera) was tested by Ficetola et al. (2021). The authors demonstrate the complexity of the marker selection process and advocate for the use of multiple markers to cover the widest range of taxa. Combining data from different markers was shown to considerably improve the match between macrobenthic indices inferred from bulk DNA and morphotaxonomic surveys (Meyer et al., 2021). A multimarker approach was also recommended for the assessment of macroinvertebrate communities from the bulk preservative (Martins et al., 2021). Despite the importance of using multiple markers, the authors also demonstrate that the presence of heavily sclerotized exoskeleton can act as a limiting factor for the detection of some taxa.The comparison of bulk DNA vs water eDNA metabarcoding has been reported by two papers. Gleason et al. (2021) show that bulk DNA metabarcoding more accurately represents the local stream macroinvertebrate community, with water eDNA data being overwhelmed by non-metazoan sequences. The same difference was observed when comparing bulk DNA to water eDNA and morphological inventories of pond macroinvertebrates (Harper et al., 2021). However, the authors consider both approaches as complementary and suggest that they should be combined for comprehensive assessment of the invertebrate community. The importance of bulk DNA metabarcoding as a tool for the assessment of marine ecosystems is also highlighted by van de Loos and Nijland (2021). The authors review various technical biases affecting bulk DNA metabarcoding workflow and discuss possible improvements that could help overcoming these biases in the future.The analysis of water samples from five sites in the Brazilian Atlantic forest and one adjacent site in Cerrado grasslands allowed Lopes et al. (2021) to demonstrate that eDNA metabarcoding significantly improves traditional monitoring methods, confirming the presence of frog species undetected by traditional methods. For a few years, invertebrate-derived DNA (iDNA) from leech blood-meal have been used to track mammalian species (Schnell et al., 2012). Here, Drinkwater et al. (2021) apply this approach to assess differences in mammalian diversity across a gradient of forest degradation in Borneo. For monitoring elusive mammals, the iDNA method complements the more traditional and widely used camera trapping.The last two papers in this section provide examples of metabarcoding optimizations aiming at improving its effectiveness in biomonitoring surveys. Guerrieri et al. (2021) show how soil preservation methods can affect estimates of taxonomic richness and community composition. The authors propose guidelines for optimizing soil preservation conditions in agreement with the objectives and practical constraints of the research project. On the other hand, Mächler et al. (2021) address the optimization of data analysis, by investigating how stringency filtering can affect eDNA diversity estimates. The authors conclude that the use of Hill numbers can help in comparisons of eDNA datasets that strongly differ in diversity.Other perspectives for eDNA-based biomonitoringThe last three articles in this special issue present ground-breaking approaches to monitoring biodiversity. Martel et al. (2021) clearly show that eDNA surveys paired with occupancy modelling can uncover metapopulation dynamics and their drivers. Such type of information is important for monitoring endangered species distributed in metapopulations and is quite difficult to obtain via traditional inventories. Shum and Palumbi (2021) reanalyzed a published marine metabarcoding dataset concerning cobble communities found within kelp forest ecosystems. They focussed on diversity data at the intraspecific level to infer population structure and demographic trends. This type of approach greatly increases the scope and value of metabarcoding studies, also opening the way towards metaphylogeography (Turon, Antich, Palacín, Praebel, & Wangensteen, 2020). Finally, Sigsgaard et al. (2021) successfully tracked insects from cow dungs from different environments, and showed that eDNA metabarcoding represents an efficient method for assessing insect diversity, with potential for biomonitoring in relation with the relatively easy standardization of such an approach.ConclusionAs shown by the collection of papers published in this issue, potential applications of eDNA in biomonitoring are highly diverse. Their scope ranges from tracking endangered species to surveying biodiversity or assessing environmental impact. Some papers focus on integrating eDNA into existing bioindication systems, whereas others use eDNA to expand the range of bioindicators and include inconspicuous, commonly overlooked microbial and meiofaunal taxa. All these papers attest to major efforts that have been done to improve eDNA methodology at every step of the workflow from sampling to data analysis. They also contribute to better understand the biological and technical factors impacting the eDNA analyses. Yet, despite this huge new knowledge and numerous practical advantages, the implementation of eDNA in routine biomonitoring still has not taken off.It is now high time to move on and to transform the eDNA field into a truly applied science. The biodiversity crisis and global environmental changes call for an urgent modernization of the tools to monitor biodiversity and assess the ecological status of our environment. As shown by the papers published here, the eDNA methodology achieved top levels of technical and scientific excellence in many areas. Certainly, there are some biases and limitations inherent to eDNA specificity, but there is no reason to consider that the technology is less “safe to use” than the conventional morpho-taxonomic approaches. There are also actions to be taken to ensure the quality and to build confidence in eDNA analyses through standardization of technical protocols and intercalibration tests. However, in view of the substantial efforts that have been made by the scientific community and illustrated by the content of this special issue, it is reasonable to expect that the implementation of eDNA-based tools in biomonitoring will not be long in coming

Aim- To describe the pattern of clinical presentation of patients with neurofibromatosis in a homogeneous black African population with emphasis on ophthalmological presentation in a multidisciplinary management setting. Methods- Ophthalmology clinic records from the Department of Ophthalmology and database of the Department of Pathology, both of University College Hopsital, Ibadan, Nigeria were reviewed for cases of neurofibromatosis over a 10-year period (Jan 2010 and Dec 2019). Relevant demographic, clinical information, management, complications and patient follow up were extracted from the records and entered into a spreadsheet and analysed. Results - The 34 cases included in this study comprised 19 males (55.9%) and age ranged from 18 months – 60 years, with a median age of 15 years. The male to female ratio was 1.3:1 with approximately 35.3% (12 patients) in the paediatric age group. The most frequent reason for consultation was unilateral progressive painless lid swelling (plexiform neurofibroma) often associated with ptosis. There was a positive family history of neurofibromatosis in 9 out of 32 cases (28.1%). Café au laît macules were observed in 22 out of 25 (88%) of cases. Typical neurofibromas were present in 84.8% of the patients. There was no significant difference in prevalence of plexiform neurofibromas with age (p= 0.05) or sex (p= 0.79). Bone and joint abnormalities was present in 17.6% of the patients. Ophthalmic examination showed conjunctiva changes in 3 cases, prominent cornea nerves in 2 cases. Iris lisch nodules was present in 75.9% of cases that had documentation, cataract in 2 cases and optic atrophy in 6cases.Three patients had ophthalmic pathway gliomas. Patients were managed by multidisciplinary teams depending on their needs. Conclusion- Multidisciplinary team management is advocated because of the multi-system disorders these patient have and the need for holistic, patient centred care that is of good quality, and sustainable.

SRY gene mutation is a common cause of 46,XY female. We report a 46,XY female with a novel mutation of SRY c.293G>A (p.Trp98ter). Our report provides evidence for a pathogenic role of the SRY gene c.293G>A mutation in an individual and enlarges the spectrum of molecular diagnosis for these patients.

Aim Viral blips that occur among virally suppressed HIV-positive patients suggest immune activation and inflammation and associated with slower CD4 count and CD4/CD8 ratio normalisation. With the advances in HIV treatment, lifestyle and comorbidities begin to be a concern despite successful antiretroviral therapy. We reported a study incorporating the effect of CD4 and CD4/CD8 ratio normalisation on viral blips in joint disease progression (DP) and time-to-event (TTE) model. Methods A total of 152 HIV-positive patients receiving efavirenz therapy were recruited. Joint DP and TTE models on viral blip were developed for CD4 and CD4/CD8 ratio separately. Risk factors, such as smoking status, pack-year and comorbidity scores, were included in the analysis. Results Gompertz model best described the CD4 and CD4/CD8 ratio DP models, while viral blips data were fitted with the Cox proportional hazard model. History of opportunistic infections and changing of antiretroviral regimen significantly affect the baseline CD4 and CD4/CD8 ratio. Comorbidity score was significant in both CD4 (asymptote CD4) and CD4/CD8 ratio DP model (recovery rate). Increase in cumulative pack-year resulted in lower CD4/CD8 ratio recovery rate (β -0.02, 95%CI: -0.03 to -0.01; p<0.001). Active smokers with slow CD4 or CD4/CD8 ratio normalisation associated with more viral blips. Conclusion CD4 and CD4/CD8 ratio are significant risk factors of viral blips and potential markers of non-AIDS related morbidities in virally suppressed patients. Early identification of high-risk group with repeated viral load testing, lifestyle modification and comorbidities management should be emphasised in the HIV treatment long-term care plan.

A 73-year-old man reported left low back pain worsening since 2 weeks. Ten days prior, he felt indefinable anxiety, which abated, but the back pain persisted. His diaphoretic and vasoconstrictive hands suggested hypovolemic shock. Emergency computed tomography revealed impending rupture of an abdominal aortic aneurysm. Emergent surgery saved his life.

Objective: To explore the effect of anxiety, depression and hypertension on quality of life (QOL) of patients with gynecological tumor and the interaction among them, and the moderating effect of social support. Design: Cross-sectional design Setting: December 2019 to July 2020, the COVID-19 outbreak phase. Population or Sample: A total of 695 gynecological cancer patients were collected. Methods: The study used the Self-Rating Anxiety Scale (SAS), the Self-Rating Depression Scale (SDS), the Functional Assessment of Cancer Therapy Genera tool (FACT-G), and the Multidimensional Scale of Perceived Social Support Scale (MSPSS). Regression analysis and the simple slope analysis were conducted. Results: QOL of patients with anxiety and hypertension had seriously deteriorated [OR=10.297, 95%CI (5.647-18.775)]. And QOL of patients with depression and hypertension also had seriously deteriorated [OR=11.846, 95%CI (6.597-21.271)]. Calculated by crossover analysis, the synergy index, attributable proportion and relative excess risk due to interact of anxiety and hypertension were1.698,0.371 and 3.822, and the correlation index of interaction between depression and hypertension were 1.475, 0.295, and 3.493. The Social Support &Depression interaction term and Social Support &Anxiety interaction term were negatively correlated with QOL (p<0.01), and explained an extra 5.7% and 5.6% of the variance respectively (p<0.01). Conclusion: Anxiety, depression and hypertension have interaction on the QOL of patients with gynecological tumors. Social support can significantly moderate the relationship between depression, anxiety and QOL respectively. Funding: National Key R&D Program of China (Grant #2018YFC1311600). Keywords: quality of life, depression, anxiety, social support, hypertension, moderating effect

All vulvar malignancies comprise only 2% of vulvar lesions, since 98% of vulvar neoplasms are benign. Nodular Fasciitis of the vulva present a diagnostic dilemma due in part to the rare nature of these disorders and similarities malignant soft tissue tumors, so publishing descriptions of these entities are essential

We report a case of a 38-year-old female patient with odontogenic myxoma who was referred to us with chief complaint of a persistent painful lesion and teeth mobility in the anterior region of the mandible which had underwent a faulty treatment plan in an outside clinic.

Here we report the management and further oral rehabilitation of a case suffering severe midface trauma following a motor vehicle accident where the patient was hit by a lorry

Human-mediated changes to natural ecosystems have consequences for both ecosystem and human health. Historically, efforts to preserve or restore ‘biodiversity’ can seem to be in opposition to human interests. However, the integration of biodiversity conservation and public health has gained significant traction in recent years, and new efforts to identify solutions that benefit both environmental and human health are ongoing. At the forefront of these efforts is an attempt to clarify ways in which biodiversity conservation can help reduce the risk of zoonotic spillover of pathogens from wild animals, sparking epidemics and pandemics in humans and livestock. However, our understanding of the mechanisms by which biodiversity change influences the spillover process is incomplete, limiting the application of integrated strategies aimed at achieving positive outcomes for both conservation and disease management. Here, we review the literature, considering a broad scope of biodiversity dimensions, to identify cases where zoonotic pathogen spillover is mechanistically linked to changes in biodiversity. By reframing the discussion around biodiversity and disease using mechanistic evidence—while encompassing multiple aspects of biodiversity including functional diversity, landscape diversity, phenological diversity, and interaction diversity—we work toward general principles that can guide future research and more effectively integrate the related goals of biodiversity conservation and spillover prevention. We conclude by summarizing how these principles could be used to integrate the goal of spillover prevention into ongoing biodiversity conservation initiatives.