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Variations of this standardised protocol have been used by Masayuki K. Sakata and colleagues to successfully extract fish eDNA from modern and historic Japanese lake and river sediments. The method has been used to recover fish species composition data from modern surface sediments from a lake (Sakata et al., 2020) and a small, natural river (Sakata et al., 2021). It has also been applied to detect target fish species in lake sediments up to 100 years old (Sakata et al., 2022). This extraction method represents a consolidation of the methods applied in the following publications: Sakata, M. K., Yamamoto, S., Gotoh, R. O., Miya, M., Yamanaka, H., & Minamoto, T. (2020). Sedimentary eDNA provides different information on timescale and fish species composition compared with aqueous eDNA. Environmental DNA, 2(4), 505– 518. https://doi.org/10.1002/edn3.75 Sakata, M. K., Watanabe, T., Maki, N., Ikeda, K., Kosuge, T., Okada, H., … Minamoto, T. (2021). Determining an effective sampling method for eDNA metabarcoding: a case study for fish biodiversity monitoring in a small, natural river. Limnology, 22(2), 221–235. https://doi.org/10.1007/s10201-020-00645-9 Sakata, M.K., Tsugeki, N., Kuwae, M., Ochi, N., Hayami, K., Osawa, R., Morimoto, T., Yasashimoto, T., Takeshita, D., Doi, H., & Minamoto, T. (2022). Fish environmental DNA in lake sediment overcomes the gap of reconstructing past fauna in lake ecosystems. bioRxiv, https://doi.org/10.1101/2022.06.16.496507

Farmed anguillid eels are frequently stocked into natural fresh waters to enhance eel resources, but little is known about what happens to these eels or their interactions with wild eels after stocking. A recent study observed a depressed survival and growth rate of farmed Japanese eels when they were reared with wild eels, which indicated that wild eels might interfere with the survival and growth of farmed-and-stocked eels through intraspecific competition. To contribute to improving eel stocking efficiency, the growth of farmed-and-stocked Japanese eels was compared among four rivers with different wild eel densities using mark-and-recapture studies. Based on the 2-year recapture survey after stocking, it was found that the density of the farmed-and-stocked eels was not significantly different among rivers. The daily growth rates of farmed-and-stocked eels in the rivers with lower wild eel density were significantly higher than those of the eels stocked into the rivers with higher wild eel density. The farmed-and-stocked eels moved significantly greater distances downstream than wild eels that showed sedentary behaviour. This and previous studies indicate that significant questions remain about the effectiveness of stocking farmed eels into water bodies where naturally recruited wild eels are present.

Patients with advanced emphysema experience breathlessness due to impaired respiratory mechanics and diaphragm dysfunction. Bronchoscopic lung volume reduction (BLVR) is a minimally invasive bronchoscopic procedure done to reduce hyperinflation and air trapping, promoting atelectasis in the targeted lobe and allowing improved respiratory mechanics. Real-world data on safety and complications outside of clinical trials of BLVR are limited.We queried the US Food and Drug Administrations (FDA) Manufacturers and User Device Experience database from May 2019 to June 2020 for reports involving BLVR with endobronchial valve (EBV) placement. Events were reviewed for data analysis.We identified 124 cases of complications during BLVR with EBV implantation. The most-reported complication was pneumothorax (110/124, 89%), all of which required chest tube placement. A total of 54 of these cases (54/110, 49%) were complicated by persistent air leak requiring additional interventions. Repeat bronchoscopy was needed to remove the valves in 28 patients, 12 were discharged with a Heimlich valve, and 10 had an additional pleural catheter placed. The other complications of BLVR with EBV placement included respiratory failure (6/124, 5%), pneumonia (4/124, 3%), hemoptysis (2/124, 1.6%), valve migration (1/124, 1%), and pleural effusion (1/124, 1%). A total of 14 deaths were reported during that year.Pneumothorax is the most-reported complication for BLVR with EBV placement, and in 65% of cases, pneumothorax is managed without removing valves. Importantly, 14 deaths were reported during that timeframe. Further studies are needed to estimate the true magnitude of the complications associated with BLVR.

Image-based profiling has emerged as a powerful technology for various steps in basic biological and pharmaceutical discovery, but the community has lacked a large, public reference set of data from chemical and genetic perturbations. Here we present data generated by the Joint Undertaking for Morphological Profiling (JUMP)-Cell Painting Consortium, a collaboration between 10 pharmaceutical companies, six supporting technology companies, and two non-profit partners. When completed, the dataset will contain images and profiles from the Cell Painting assay for over 116,750 unique compounds, over-expression of 12,602 genes, and knockout of 7,975 genes using CRISPR-Cas9, all in human osteosarcoma cells (U2OS). The dataset is estimated to be 115 TB in size and capturing 1.6 billion cells and their single-cell profiles. File quality control and upload is underway and will be completed over the coming months at the Cell Painting Gallery:https://registry.opendata.aws/cellpainting-gallery. A portal to visualize a subset of the data is available athttps://phenaid.ardigen.com/jumpcpexplorer/.

Environmental DNA (eDNA) analysis allows noninvasive and cost-effective monitoring of macroorganisms' distribution and composition in aquatic ecosystems. Benzalkonium chloride (BAC) is an inexpensive and simple preservative for eDNA in water samples and has been used in many eDNA studies for preventing its degradation during transportation. However, its preservation performance has limitedly been evaluated by species-specific assays, targeting short fragments of mitochondrial DNA in freshwater and brackish ecosystems. Here, we examined the performance of BAC in preserving eDNA in seawater samples, targeting different fragment lengths of mitochondrial and nuclear eDNA, and community information inferred by eDNA metabarcoding. We quantified the time series changes of Japanese jack mackerel (Trachurus japonicus) eDNA concentrations in experimental tanks and inshore seawater to compare the yields and decay rates of eDNA between BAC treatments. For both tank and field samples, BAC treatment substantially suppressed the degradation of all types of target eDNA and increased the eDNA yields at the start of the experiment. In addition, we performed eDNA metabarcoding targeting fish community to compare the species richness and composition in seawater samples between BAC treatments, showing that the number of fish species in field samples hardly varied throughout a day by BAC treatment. Our findings indicate high versatility of BAC in preserving both the quantitative (copy number) and the qualitative (species richness) information on various types of aqueous eDNA in various environmental conditions. BAC should therefore be used to minimize the false-negative detection of eDNA, regardless of target genetic regions, fragment sizes, environmental conditions, and detection strategies.

Background Genetic testing for inherited arrhythmias and discriminating pathogenic from benign variants are integral for the gene-based medicine. However, the high throughput in vivo functional analysis for the rare variants of the KCNQ1 potassium channel is scarce. Purpose We tested the utility of the in vivo zebrafish cardiac assay for determining the pathogenicity of the KCNQ1 variants identified in patients with long QT syndrome (LQTS) and atrial fibrillation (AF). Methods We generated a knock-out zebrafish with CRISPR-mediated insertions or deletions of the KCNQ1 homolog in zebrafish (kcnq1+/+). To test the utility of the cardiac assay, we used five KCNQ1 variants identified in patients with LQTS or familial AF. Human wild-type or mutant KCNQ1 cRNA (Q1) was co-injected with human KCNE1 cRNA (E1) into the F3 generation embryos with homozygous deletions. We dissected the hearts from the thorax at 72 hour-post-fertilization and measured transmembrane potential in zebrafish heart using the disrupted patch technique. Action potential duration was calculated as the time interval between the peak maximum upstroke velocity and 90% of repolarization (APD90). We compared the APD90s with patients' clinical phenotype and IKs density measured by patch-clamp technique in heterologous system. Results The mean APD90 of embryos with kcnq1del/del was 279±48 ms, which was restored by injecting Q1 WT and E1 (159±29 ms) to that with kcnq1+/+ (167±28 ms). We tested if the mean APD90 of embryos with kcnq1del/del was restored (shortened) by injecting the KCNQ1 variants. First we tested the dominant negative variant p.S277L and the trafficking deficient variant p.T587M. Patients with these variants showed significant prolonged QT intervals, and patch clamp study showed both variants caused the non-functional channels. Zebrafish cardiac assay showed the mean APD90 of embryos with kcnq1del/del+ Q1 S277L+E1 or Q1 T587M+E1 was significantly longer than that with kcnq1del/del+Q1 WT+E1 (Table). Next we tested in-frame variant c.1472_1473 ins GGACCT, which was identified from a patient with AF and normal QT interval. Patch clamp study showed the current density of the mutant KCNQ1 channel with KCNE1 was comparable to that of wild-type KCNQ1 channel with KCNE1. Zebrafish assay showed the mean APD90 of embryos with kcnq1del/del shortened by injecting Q1 insACCTGG +E1 (Table). Finally we tested a missense variant p.R451Q, which was identified from a patient with LQTS. Patch clamp study showed the currents in the cells transfected with R451Q+KCNE1 were similar to those with WT+KCNE1. Zebrafish assay showed the mean APD90 of embryos with kcnq1del/del+Q1 R451Q+E1 was longer than that with kcnq1del/del+Q1 WT+E1 (Table). Conclusions Functional analysis of in vivo zebrafish cardiac assay might be useful for determining the pathogenicity of rare variants in patients with LQTS. Funding Acknowledgement Type of funding sources: Public Institution(s). Main funding source(s): The Grant-in-Aid for Scientific Research (C)

Recently, environmental DNA (eDNA) metabarcoding techniques have been applied to biodiversity investigations in aquatic ecosystems. However, no study has yet tested whether this technique is effective for water bodies in which extensive algal blooms break out. In this study, fish eDNA metabarcoding was carried out in Lake Taihu, which experiences extensive algal blooms, to confirm whether the technique is also effective for fish diversity research in ecosystems with frequent and extensive blooms. In December 2016, three samples were collected, including one collected in the presence of algal blooms and two collected in the absence of algal blooms. In August 2017, six samples were collected, including three collected in the presence of algal blooms and three in the absence of algal blooms. Equal amount of water samples (1 L) was collected from each site; however, the actual amount of filtrate varied with the site. Twenty-seven freshwater fish species were detected from the water samples collected in Lake Taihu. The results showed that the composition of the detected species did not differ whether or not blooms were present. However, the amount of filtration could influence the number of species detected. The results suggest that future eDNA metabarcoding studies under similar water environments should increase the amount of filtration to maximize number of species detected.

No abstract available. Article truncated after 150 words. A 58-year-old woman with a history of orthotopic heart transplant, performed for Adriamycin-induced cardiomyopathy, treated with mycophenolate and tacrolimus, presented for routine interval follow up. The patient’s past medical history was significant for follicular thyroid carcinoma treated with total thyroidectomy and bilateral breast carcinoma in remission as well as hypothyroidism and type II diabetes mellitus. In addition to tacrolimus and mycophenolate, the patient’s medications included aspirin, insulin, itraconazole (for anti-fungal prophylaxis), levothyroxine, prednisone (tapering since transplant), and valganciclovir. The patient recently complained of rhinorrhea and cough productive of brown-tinged sputum, improving over the previous 2 weeks; she denied fever, chills, shortness of breath, night sweats chest pain, or gastrointestinal symptoms. Physical examination showed the patient to be afebrile with normal heart and respiratory rates and blood pressure. Her room air oxygen saturation was 99%. The patient’s complete blood count and serum chemistries showed largely normal values, with the white blood …

Underwater sediments are a natural archive of biological information. Reconstruction of past fauna has been conducted for various taxonomic groups using morphological remains and DNA derived from these remains. However, information on past occurrences of fish species, the top predator of lake ecosystems, could have been reproduced only in exceptional environments, and past quantitative information on fish, particularly in lake ecosystems, has been a knowledge gap in reconstructing past fauna. Tracking the quantitative fluctuations of fish is essential for reconstructing multiple trophic levels of organisms in lake ecosystems.To acquire past quantitative fish information from lake sediments, we collected approximately 30 cm-length of underwater sediments in Lake Biwa. We extracted sedimentary environmental DNA (eDNA) and measured temporal fluctuations in the eDNA concentration of the native and fishery target species Plecoglossus altivelis and Gymnogobius isaza. For P. altivelis, we examined the possibility of tracking quantitative fluctuations by comparing sedimentary eDNA with recorded catch per unit effort (CPUE).The chronology of the sediments allowed us to obtain information on sediments collected in Lake Biwa over the past 100 years. The deepest depths at which sedimentary eDNA was detected were 30 and 13 cm for P. altivelis and G. isaza from the surface, corresponding to approximately 100 and 30 years ago, respectively. In the comparison of sedimentary eDNA concentrations and biomass, we found a significant correlation between the CPUE of P. altivelis and its sedimentary eDNA concentration adjusted to compensate for DNA degradation. Sedimentary eDNA fluctuations were observed in P. altivelis, possibly reflecting the abundance fluctuation due to variations in the main food resources of zooplankton.Our findings provide essential pieces for the reconstruction of past fauna of lake ecosystems. The addition of quantitative information on fish species will reach a new phase, for instance, by investigating population shifts or biological interactions in the reconstruction of past fauna in lake ecosystems.

This Personal Account describes the authors' foray into DNA-encoded libraries. The article addresses several key aspects of this hit generation technology, from the development of new synthetic methodology to the subsequent conception, design, and delivery of a DNA-encoded library. In particular, we have been engaged in adapting photocatalytic reactions to the idiosyncratic requirements of DNA-encoded chemistry. We have chosen one such methodology, namely a photocatalytic [2+2] cycloaddition reaction, to showcase how we employed property-based computational analyses to guide the selection and validation of building blocks for the production of a library. Ultimately, these novel bond disconnections and design principles led to the assembly of a DNA-encoded library that is composed of structurally diverse compounds within largely desirable property space and, therefore, well positioned to deliver novel chemical matter for drug discovery programs.

We describe a simple and robust procedure for the Rh(III)-catalyzed [4+2] cycloaddition of feedstock gases enabled through C–H activation. A diverse set of 3,4-dihydroisoquinolones and 3-methylisoquinolones have been prepared in good to excellent yields. The effects of using ethylene and propyne as coupling partners on C–H site selectivity have also been explored with a representative set of substrates and are discussed herein.

In recent years, biodiversity loss has become one of the most serious environmental issues worldwide, especially in aquatic ecosystems. To avoid diversity loss, it is necessary to monitor biological communities, and environmental DNA (eDNA) metabarcoding has been developed as a rapid, noninvasive, and cost-effective method for aquatic biodiversity monitoring. Although this method has been applied to various environments and taxa, a detailed assessment of the efficient sampling methods for monitoring is still required. In this study, we explored eDNA metabarcoding sampling methods for fish at a single site to maximize the number of detected species using realistic effort in a natural, small river. We considered the following three parameters: sample type (water or sediment), sample position at a site (right and left shore and center of the river), and water volume (10–4000 mL). The results suggested that the number of detected species from sedimentary eDNA was equivalent to that from aqueous eDNA, although the species composition was different. The number of detected species could be saturated by collecting a 1000 mL water sample, regardless of sampling position within a survey site. However, sedimentary eDNA showed a spatially heterogeneous species composition between sampling positions within a survey site despite the short distance (5 m) between positions, without apparent differences in physical properties such as velocity and sediment particle distribution. By completing eDNA biodiversity monitoring of fish with 1000 mL water samples across the whole river, we detected more fish species than in previous traditional surveys conducted at the same sites. Thus, the aqueous eDNA metabarcoding method is as efficient as traditional surveys, while sedimentary eDNA metabarcoding could complement the results of aqueous eDNA metabarcoding.

In the present work, attempts of reducing a graphene oxide powder using a low temperature hydrogenation disproportionation desorption and the recombination process (L-HDDR) has been carried out. A lower processing temperature in large scale production is significant when costs are concerned. Graphite oxide was prepared using a modified Hummers’ method dispersed in ethanol and exfoliated using ultrasonication to produce Graphene Oxide (GO). Investigations have been carried out by X-ray diffraction (XRD) and scanning electron microscopy (SEM). The experimental results of L-HDDR processing graphene oxide powder, using unmixed hydrogen at 400°C and relatively low pressures (

Objective: To assess the contribution and safety of bronchoscopic cryobiopsy vs traditional forceps biopsy used in clinical practice for diagnosing diffuse parenchymal lung disease (DPLD). Patients and Methods: We identified 271 patients who underwent bronchoscopic biopsy for DPLD at Mayo Clinic, MN (June 1, 2013, through September 30, 2017). Medical records were reviewed including prebiopsy clinical and radiographic impressions. Diagnostic yield was assessed in terms of a specific histologic pattern resulting in a diagnosis when combined with the clinical-radiologic context. Clinical utility was defined as a biopsy result deemed useful in patient management. Results: The cohort included 120 cryobiopsy and 151 forceps biopsy cases with mean age 61±14 years and 143 (53%) men. Diagnostic yield (55% vs 41%; odds ratio [OR], 1.73; 95% CI, 1.07 to 2.83; P=.026) and clinical utility (60% vs 40%; OR, 2.21; 95% CI, 1.36 to 3.63; P=.001) were higher for the cryobiopsy group, and the association remained after control for prebiopsy clinical impressions (OR, 2.21; 95% CI, 1.22 to 4.08; P=.010 and OR, 3.23; 95% CI, 1.76 to 6.10; P

Aqueous environmental DNA (eDNA) analysis has been applied to the monitoring of various ecosystems and taxa, and the characteristics of aqueous eDNA have been previously studied. In contrast, although sedimentary eDNA has been used to restore past information, the characteristics of sedimentary eDNA are not well understood. In this study, we compared the properties of sedimentary and aqueous eDNA of macro‐organisms. First, to clarify the preservation ability of sediments, we compared the difference in decay rates between aqueous and sedimentary eDNA using samples collected from a biotope (an artificial pond prepared with concrete). Next, to clarify the biological information retained in sedimentary eDNA both qualitatively and quantitatively, we compared eDNA concentrations between sediment and water samples collected simultaneously from a lake, and the fish species detected by eDNA metabarcoding were also compared. The results demonstrated the following: (a) the decay rate (decreased eDNA copy number divided by the initial eDNA copy number per unit time) of sedimentary eDNA (0.00033 ± 0.000049 [mean ± SE]/hr) was lower than that of aqueous eDNA (0.01863 ± 0.0011/hr); (b) sedimentary eDNA concentration of the mitochondrial marker of three fish species was higher than aqueous eDNA concentration for the same sample weight (12.5–1,456.9 times); and (c) the species composition obtained by metabarcoding was not significantly different between sediment and water; however, considering the lower decay rate of sedimentary eDNA, using both sample types may provide more comprehensive information of species distribution. Thus, sedimentary eDNA analysis will expand future biomonitoring and ecological studies by providing a difference in timescale.

Prescription opioids are a major cause of the opioid epidemic. Despite the minimally invasive nature of medical thoracoscopy (MT), data on the efficacy of non-opioid-based pain control after MT is lacking. The purpose of this study is to assess the feasibility and efficacy of a non-opioid-based pain management strategy in patients who underwent MT.We performed a retrospective analysis of all patients who underwent MT in the Mayo Clinic (Minnesota and Arizona) outpatient setting. We assessed their pain level and the need for analgesia post-MT from August 1, 2019, to May 24, 2021.Forty patients were included. In the first 24 hours, 5/40 (12.5%) reported no pain. Twenty-eight patients out of 40 (70%) reported minor pain (pain scale 1-3), and 7/40 (17.5%) reported moderate pain (pain scale 4-6). No patients reported severe pain. Twenty-two out of 35 patients who experienced discomfort (63%) required acetaminophen, 6/35 patients (17%) required nonsteroidal anti-inflammatory drug, and 7/35 patients (20%) did not require analgesia. Of the 7 patients who had moderate pain, 5 (71%) reported that the moderate pain improved to mild at 72 hours post-MT. Zero patients required opioids, and none reported contacting any provider to manage the pain post-MT. Fourteen patients (78%) who had both parietal pleural biopsies and tunneled pleural catheter placed reported minor pain, 3 patients (17%) reported moderate pain, and 1 patient (6%) experienced no discomfort.MT is well-tolerated by patients with non-opioid-based pain management strategy as needed if there is no absolute contraindication.

Biodiversity monitoring is important for the conservation of natural ecosystems in general, but particularly for amphibians, whose populations are pronouncedly declining. However, amphibians’ ecological traits (e.g. nocturnal or aquatic) often prevent their precise monitoring. Environmental DNA (eDNA) metabarcoding – analysis of extra-organismal DNA released into the environment – allows the easy and effective monitoring of the biodiversity of aquatic organisms. Here, we developed and tested the utility of original PCR primer sets. First, we conducted in vitro PCR amplification tests with universal primer candidates using total DNA extracted from amphibian tissues. Five primer sets successfully amplified the target DNA fragments (partial 16S rRNA gene fragments of 160–311 bp) from all 16 taxa tested (from the three living amphibian orders Anura, Caudata and Gymnophiona). Next, we investigated the taxonomic resolution retrieved using each primer set. The results revealed that the universal primer set “Amph16S” had the highest resolution amongst the tested sets. Finally, we applied Amph16S to the water samples collected in the field and evaluated its detection capability by comparing the species detected using eDNA and physical survey (capture-based sampling and visual survey) in multiple agricultural ecosystems across Japan (160 sites in 10 areas). The eDNA metabarcoding with Amph16S detected twice as many species as the physical surveys (16 vs. 8 species, respectively), indicating the effectiveness of Amph16S in biodiversity monitoring and ecological research for amphibian communities., 水をくむだけの新しい両生類の調査法. 京都大学プレスリリース. 2022-02-22.

Environmental DNA (eDNA) analysis has seen rapid development in the last decade, as a novel biodiversity monitoring method. Previous studies have evaluated optimal strategies, at several experimental steps of eDNA metabarcoding, for the simultaneous detection of fish species. However, optimal sampling strategies, especially the season and the location of water sampling, have not been evaluated thoroughly. To identify optimal sampling seasons and locations, we performed sampling monthly or at two‐monthly intervals throughout the year in three dam reservoirs. Water samples were collected from 15 and nine locations in the Miharu and Okawa dam reservoirs in Fukushima Prefecture, respectively, and five locations in the Sugo dam reservoir in Hyogo Prefecture, Japan. One liter of water was filtered with glass‐fiber filters, and eDNA was extracted. By performing MiFish metabarcoding, we successfully detected a total of 21, 24, and 22 fish species in Miharu, Okawa, and Sugo reservoirs, respectively. From these results, the eDNA metabarcoding method had a similar level of performance compared to conventional long‐term data. Furthermore, it was found to be effective in evaluating entire fish communities. The number of species detected by eDNA survey peaked in May in Miharu and Okawa reservoirs, and in March and June in Sugo reservoir, which corresponds with the breeding seasons of many of fish species inhabiting the reservoirs. In addition, the number of detected species was significantly higher in shore, compared to offshore samples in the Miharu reservoir, and a similar tendency was found in the other two reservoirs. Based on these results, we can conclude that the efficiency of species detection by eDNA metabarcoding could be maximized by collecting water from shore locations during the breeding seasons of the inhabiting fish. These results will contribute in the determination of sampling seasons and locations for fish fauna survey via eDNA metabarcoding, in the future., Number of detected species by eDNA metabarcoding for (a) Miharu dam reservoir, (b) Okawa dam reservoir, and (c) Sugo dam reservoir. The number of species detected by eDNA survey peaked in May in Miharu and Okawa reservoirs, and in March and June in Sugo reservoir, which corresponds with the breeding seasons of many of the fish species inhabiting the reservoirs.

[Image: see text] Two novel compounds were identified as Naa50 binders/inhibitors using DNA-encoded technology screening. Biophysical and biochemical data as well as cocrystal structures were obtained for both compounds (3a and 4a) to understand their mechanism of action. These data were also used to rationalize the binding affinity differences observed between the two compounds and a MLGP peptide-containing substrate. Cellular target engagement experiments further confirm the Naa50 binding of 4a and demonstrate its selectivity toward related enzymes (Naa10 and Naa60). Additional analogs of inhibitor 4a were also evaluated to study the binding mode observed in the cocrystal structures.

Monitoring reproductive migration is essential for the conservation of anadromous species. Shishamo smelt (Spirinchus lanceolatus) is endemic to Hokkaido, the northernmost large island in Japan. S. lanceolatus is an anadromous species that is known to migrate into rivers for a very short period in early winter. While this species has a special value for local fisheries, the catch amount has drastically declined in the last few decades. Information about S. lanceolatus reproductive migration dynamics is limited, which prevents them from being efficiently managed as a resource. In this study, we used environmental DNA (eDNA) methods as a noninvasive molecular tool for estimating presence/absence and abundance/biomass of S. lanceolatus during their migration into rivers. We developed a species‐specific qPCR system for S. lanceolatus, examining (a) temporal variation in S. lanceolatus eDNA concentrations compared with catch data gathered by traditional methods and (b) variability of migratory patterns among river systems. In a core river for their spawning migration, we consistently detected S. lanceolatus eDNA throughout the spawning season, and the temporal distribution of eDNA concentration was consistent with that of the number of migrating S. lanceolatus estimated by catch survey data. In addition, we were able to detect S. lanceolatus eDNA even from rivers without any official record of their migration. Among rivers with eDNA detection, the relative eDNA concentrations varied, indicating that the population biomass differs largely among the river populations. Our study suggests that eDNA detection systems are useful for tracking reproductive migration of S. lanceolatus at fine spatio‐temporal scales.

Traditional methods to incorporate metals into graphene oxide (GO) usually require toxic reagents or high temperatures. This study proposes an innovative and sustainable method to incorporate silver (Ag) into graphene oxide using electron beam and evaluate its antibacterial activities. The method is based on green synthesis, without toxic reagents or hazardous wastes, and can be carried out at room temperature, in short reaction times. To synthesize the Ag/rGO nanocomposite, a water/isopropanol solution with dispersed graphene oxide and silver nitrate was submitted to a dose range from 150 to 400 kGy. The product was characterized by thermogravimetry analysis, X-ray diffraction and transmission electron microscopy. The antibacterial activity of Ag/rGO was observed against Gram-negative Escherichia coli by plate count method and atomic force microscopy. The results showed that concentrations as low as 100 μg/mL of produced Ag/rGO were enough to inactivate the cells.

Introduction: Preventing mosquito-borne infectious diseases requires that vector mosquitoes are monitored and controlled. Targeting immature mosquitoes (eggs, larvae, and pupae), which have less mobility than adults, is an effective management approach. However, conducting these surveys is often difficult due to the limitations of morphological classification and survey costs. The application of environmental DNA (eDNA) analysis can solve these issues because it allows easy estimation of species distribution and morphology-independent species identification. Although a few previous studies have reported mosquito eDNA detection, there is a gap in knowledge regarding the dynamics of mosquito eDNA during the developmental stages of immature mosquitoes. Methods: We used Culex pipiens pallens, a vector of West Nile fever, as a model species. First, we developed a species-specific detection assay and confirmed its specificity using in silico and in vitro tests. Next, we conducted laboratory experiments using breeding tanks. Water samples were collected at each developmental stage. In addition, water samples were collected daily until the seventh day after emergence from the pupae. We quantified eDNA using real-time PCR with the developed assay to investigate the dynamics of mosquito eDNA. Results: The specificity of the developed assay was confirmed by in silico and in vitro tests. Mosquito eDNA was detected at all developmental stages and detected up to seven days after emergence of pupae. In particular, high concentrations of eDNA were detected immediately after hatching from eggs and after emergence from pupae. Highly frequent positive eDNA signals were continuously detected between egg hatching and pupa hatching. Conclusions: Mosquito eDNA was detected immediately after the eggs were introduced, and eDNA-positive detections continued until pupae emergence, suggesting that eDNA analysis is useful for monitoring mosquito larvae. The results show that eDNA analysis provides valuable information about the water sources inhabited by immature mosquitoes in mosquito control. In the future, monitoring immature mosquitoes using eDNA analysis will aid in preventing mosquito-borne infectious diseases.

The spread of antimicrobial-resistant bacteria (ARB) in natural environments including wild animals is a concern for public health. Birds cover large areas, and some fly across borders to migrate in large flocks. As a migratory bird, the Greater White-fronted Goose (Anser albifrons) travels to Miyajimanuma, North Japan, each spring and autumn. To investigate the ARB in migratory birds and their surroundings, we collected 110 fecal samples of A. albifrons and 18 water samples from Miyajimanuma in spring and autumn of 2019. Isolation of Escherichia coli was performed using selective agars with or without antimicrobials (cefazolin and nalidixic acid). Isolates of E. coli were recovered from 56 fecal samples (50.9%) and five water samples (27.8%) on agars without antimicrobials. No isolates were recovered on agars with antimicrobials. One E. coli isolate derived from a fecal sample exhibited resistance to β-lactams (ampicillin and cefazolin), whereas all other isolates exhibited susceptibility to all tested antimicrobials. The resistant isolate harbored blaACC, which could be transferred to other bacteria and confer resistance to β-lactams. These results suggest a low prevalence of antimicrobial resistance in wild migratory birds and their living environments; however, wild migratory birds sometimes carry ARB harboring transferrable antimicrobial resistance genes and therefore present a risk of spreading antimicrobial resistance.

Background Increased arterial stiffness characterize by aging. It is reported that age-related increases in muscle sympathetic nerve activity (MSNA) may be one of contributing factor for arterial stiffness. Arterial reflected wave was composed of SNA and aging. Increased arterial reflected wave partly plays an important role in blood pressure. Recently, we reported that arterial velocity pulse index (AVI), a novel index of arterial reflected waves, was associated with MSNA in hypertensive patients. It is still uncertain the effect of age on the association between AVI and SNA in hypertensive patients. Method Patients with essential HT and matched non-hypertensive control subjects were included in this study. HT was diagnosed as systolic blood pressure (SBP) ≥140 mmHg or diastolic blood pressure (DBP) ≥90 mmHg. AVI was measured from left upper arm by NAS-1000. SNA was evaluated by direct recording of muscle sympathetic nerve activity (MSNA) from peroneal nerves. Results 45 HT patients and 46 control subjects were included. Age, SBP and DBP were significantly increased in HT group compared to control (Age 63±14 vs 42±16 years, p160 mmHg subjects in group 2, significant correlation was clarified between AVI and MSNA (r=0.62, p Conclusion The relationship between AVI and MSNA in HT patients is preserved regardless of aging, however, high blood pressure over 160mmHg might obscure its correlation. These results indicate that AVI is useful to estimate sympathetic nerve activity in high aging HT patient treated Funding Acknowledgement Type of funding sources: None.

Introduction Left ventricular diastolic dysfunction (LVDD) is the main cause of heart failure with preserved ejection fraction (HFpEF). LVDD is related not only to arterial stiffness but also sympathetic nerve activity (SNA). Recent study demonstrated that increased muscle sympathetic nerve activity (MSNA) may be one of contributing factor for arterial stiffness. In clinical practice, Cardio-ankle vascular index (CAVI) provides a reproducible index of arterial stiffness, independent of blood pressure (BP). Recently, Arterial Velocity pulse Index (AVI), which is an index of arterial reflected waves, have been proposed as new index of arterial stiffness. We reported that AVI was associated with MSNA in hypertensive (HT) patients. However, it is still uncertain the effect of LVDD on the association between AVI and SNA in HT patients. Thus, we tested the hypothesis that AVI would be increased and related to MSNA in HT patients with LVDD. Methods Patients with essential HT subjects were included in this study. HT was diagnosed as systolic blood pressure (SBP) ≥140mmHg or diastolic blood pressure (DBP) ≥90mmHg. Patients with secondary HT was excluded. AVI was measured from left upper arm by NAS-1000 (Nihon Koden, Japan). CAVI was measured by VaSera VS-1500A (Fukuda Denshi, Japan). Transthoracic echocardiography was performed by trained sonographers. SNA was evaluated by direct recording of MSNA from peroneal nerves. Results 25 HT patients were included (age 63±14 years, Male/Female 9/16). They were divided into two groups according to E/e' (no LVDD group, E/e' ≤9, N=12; LVDD group, E/e' >9, N=13). There were no significant differences between no LVDD and LVDD groups in age (63±9 vs 69±9 years p=0.205), body mass index (23±3 vs 24±4 p=0.355), BP (SBP 139±16 vs 144±20mmHg p=0.524, DBP 87±15 vs 78±14mmHg p=0.167). LV Ejection Fraction (EF) and Stroke Volume (SV) did not differ between two groups (EF 66±7 vs 69±6% p=0.471, SV 58±7 vs 62±14ml p=0.599). MSNA had tendency to increase in LVDD group compared to no LVDD group (MSNA 53±10 vs 44±12 bursts/100 heartbeats, p=0.052). Contrary to our hypothesis, AVI and CAVI did not differ between two groups (AVI 27±7 vs 29±7 p=0.398, CAVI 8.7±1.4 vs 8.6±1.4 p=0.894). However, a significant correlation was seen between AVI and MSNA in no LVDD group (r=0.57, p Conclusion AVI was significantly associated with MSNA in HT patients without LVDD, but not with LVDD. CAVI was related to AVI in HT patients with LVDD, but not without LVDD. MSNA was slightly increased in HT patients with LVDD compared to without LVDD. These results indicate that augmented SNA could contribute to the increase in arterial stiffness in HT patients without LVDD, however, this contribution might be attenuated in HT patients with LVDD. Funding Acknowledgement Type of funding sources: None.

Interventional radiofrequency (RF) ablation techniques are indicated when an adequate effect is not obtained with conservative measures.The primary objective of this study was to evaluate pain relief after RF denervation of the sacroiliac joint. The secondary objective was to evaluate pain intensity and relief duration.The study was retrospective.The study was conducted at Vera Cruz Hospital, Campinas, Brazil.Data were collected from the medical records of patients undergoing RF denervation for low back pain originating in the sacroiliac joint, from January 2015 to December 2017. There were 78 patients studied, between 18 and 65 years old, of both genders, ASA I or II, who underwent knee arthroscopic meniscectomy. The patients were submitted to denervation of sacroiliac joint by 3 types of RF (conventional, pulsed, and cooled). The following parameters were evaluated, number of patients who obtained ? 50% pain relief; pain intensity, measured using the visual analog scale (before the procedure and 15, 30, 90 and 180 days after, performed by the same evaluator); and the use of complementary analgesic for 2 weeks.Of the 78 included patients, 56 (71.8%) underwent conventional RF, 9 (11.5%) underwent pulsed RF, and 13 (16.7%) underwent cooled RF. There were losses to follow-up including 40 patients who underwent conventional RF, 5 who underwent pulsed RF, and 12 who underwent cooled RF, who were retained for 6 months. There was significant pain relief with the three types of RF for up to 6 months of follow-up, with no difference among the types. After 6 months, 90.2% of patients who underwent conventional RF, 100% who underwent pulsed RF, and 91.7% who underwent cooled RF maintained ≥ 50% pain relief. Complementary analgesics were used by 95% of the patients who underwent conventional RF, 80% who underwent pulsed RF, and 91% who underwent cooled RF 2 weeks after the procedure. There were mild adverse effects, such as edema, hematoma, and local pain, without complications.As for limitations, the number of pulsed and cooled RF is low and in a retrospective study some data may be missing, especially from follow-up.RF denervation of the sacroiliac joint is effective and promotes a long-lasting analgesic effect.

Preventing mosquito-borne infectious diseases requires that vector mosquitoes are monitored and controlled. Targeting immature mosquitoes (eggs, larvae, and pupae), which have less mobility than adults, is an effective management approach. However, conducting these surveys is often difficult due to the limitations of morphological classification and survey costs. The application of environmental DNA (eDNA) analysis can solve these issues because it allows easy estimation of species distribution and morphology-independent species identification. Although a few previous studies have reported mosquito eDNA detection, there is a gap in knowledge regarding the dynamics related to the persistence of immature mosquito eDNA. We used Culex pipiens pallens, a vector of West Nile fever, as a model species. First, we developed a species-specific detection assay and confirmed its specificity using in silico and in vitro tests. Next, we conducted laboratory experiments using breeding tanks. Water samples were collected at each developmental stage. In addition, water samples were collected daily until the seventh day after emergence from the pupae. We quantified eDNA using real-time PCR with the developed assay to investigate the dynamics of mosquito eDNA. The specificity of the developed assay was confirmed by in silico and in vitro tests. Mosquito eDNA was detected at all developmental stages and detected up to seven days after emergence of pupae. In particular, high concentrations of eDNA were detected immediately after hatching from eggs and after emergence from pupae. Highly frequent positive eDNA signals were continuously detected between egg hatching and pupa hatching. Mosquito eDNA was detected immediately after the eggs were introduced, and eDNA-positive detections continued until pupae emergence, suggesting that eDNA analysis is useful for monitoring mosquito larvae. In the future, monitoring immature mosquitoes using eDNA analysis will contribute to prevent mosquito-borne infectious diseases.

Alien ant species (Formicidae, Hymenoptera) cause serious damage worldwide. Early detection of invasion and rapid management are significant for controlling these species. However, these attempts are sometimes hindered by the need for direct detection techniques, such as capture, visual observation, or morphological identification. In this study, we demonstrated that environmental DNA (eDNA) analysis can be used as a monitoring tool for alien ants using Linepithema humile (Argentine ant), one of the most invasive ants, as a model species. We designed a new real-time PCR assay specific to L. humile and successfully detected eDNA from the surface soil. The reliability of eDNA analysis was substantiated by comparing eDNA detection results with traditional survey results. Additionally, we examined the relationship between eDNA concentration and distance from nests and trails. Our results support the effectiveness of eDNA for alien ant monitoring and suggest that this new method could improve our ability to detect invasive ant species.

Musculoskeletal involvement is one of the major causes of impairment in daily life of patients with systemic sclerosis (SSc). Several hand radiographic findings can be seen in SSc patients; however, their association with clinical features and autoantibodies remains unclear. Here, we analyzed 124 SSc patients with their hand X-rays and clinical and serological features. Abnormal findings in hand X-rays including acro-osteolysis, calcinosis, flexion contracture, erosive change, joint space narrowing, and subluxation were observed in 110 patients (68%). These X-ray findings were more prevalent in patients with longer disease duration and digital ischemic changes. The majority of erosions were typical for erosive hand osteoarthritis, which was seen in 19% of patients. Hand X-ray findings were associated with involved organs; acro-osteolysis with interstitial lung disease, calcinosis with pulmonary arterial hypertension and gastrointestinal tract involvement, and flexion contracture with gastrointestinal tract involvement. Those findings were also relevant to autoantibodies; acro-osteolysis was more common in SSc patients with anti-Scl70 antibody but less in patients with anticentromere antibody. Calcinosis was more prevalent in patient with anticentromere antibody. In our study, organ involvements and SSc-associated autoantibodies showed associations with hand radiographic abnormalities. Hand X-ray findings might reflect underlying pathogenesis and autoantibody profiles in SSc patients.Key Points• Hand X-ray abnormalities were observed in approximately two-thirds of patients with SSc.• Erosive osteoarthritis was more prevalent in SSc patients than general population.• Hand X-ray findings were associated with disease duration, organ involvements, and SSc-associated autoantibodies, reflecting underlying pathogenesis.