Synthesis of adipic acid (AA) through the oxidation of cyclohexanol and cyclohexanone (K/A oil) with nitric acid was conducted in a capillary microreactor system. Effects of the temperature, the nitric acid concentration, the volumetric flow rate ratio of nitric acid to K/A oil, and the capillary length on the selectivity and the product yield were investigated systematically to achieve optimal reaction conditions. Notably, a high yield of adipic acid (i.e., 90%) was achieved just in 6 seconds at 85℃ with the use of 55 wt% nitric acid. Gas components produced in this oxidation process and its total volumetric flow rate were determined under various operating conditions, which was beneficial for reaction mechanism characterization and process optimization. Finally, a kinetic model was established, which was of crucial theoretical significance and practical value for optimizing the reactor design and better understanding such fast and highly exothermic multiphase processes with abundant gas production.

Background Charge density mapping of atrial fibrillation (AF) reveals dynamic patterns of localised rotational activation (LRA), irregular activation (LIA) and focal firing (FF). Their spatial stability, conduction characteristics and the optimal duration of mapping required to reveal these phenomena and has not been explored. Methods Bi-atrial mapping of AF propagation was undertaken and variability of activation patterns quantified up to a duration of 30-seconds(s). The frequency of each pattern was quantified at each vertex of the chamber over 2 separate 30s recordings prior to ablation and R2 calculated to quantify spatial stability. Regions with the highest frequency were identified at increasing time durations and compared to the result over 30s using Cohen’s kappa. Properties of regions with the most stable patterns were assessed during sinus rhythm and extrastimulus pacing. Results In twenty-one patients, 62 paired LA and RA maps were obtained. LIA was highly spatially stable with R2 between maps of 0.83(0.71-0.88) compared to 0.39(0.24-0.57) and 0.64(0.54-0.73) for LRA and FF, respectively. LIA was also most temporally stable with a kappa of >0.8 reached by 12s. LRA showed greatest variability with kappa>0.8 only after 22s. Regions of LIA were of normal voltage amplitude (1.09mv) but showed increased conduction heterogeneity during extrastimulus pacing (p=0.0480). Conclusion Irregular activation patterns characterised by changing wavefront direction are temporally and spatially stable in contrast with rotational patterns that are transient with least spatial stability. Focal activation appears of intermediate stability. Regions of LIA show increased heterogeneity following extrastimulus pacing and may represent fixed anatomical substrate.

Viral vectors have a great potential for gene delivery, but manufacturing at pharmaceutical scale is a big challenge for the industry. The baculovirus-insect cells system is one of the most scalable platforms to produce clinical grade recombinant Adeno-Associated Virus (rAAV) vectors, however, the standard procedure to generate recombinant baculovirus based on Tn7 transposition is time consuming and still suffers technical constraints. Indeed, we recently shown that baculoviral sequences adjacent to the AAV ITRs are preferentially encapsidated into the rAAV vector particles. This observation raised concern about safety for clinical applications due to the presence of bacterial and antibiotic resistance coding sequences with Tn7-mediated system for the construction of recombinant baculoviruses. Here, we investigated a faster and safer method to generate baculovirus reagents based on homologous recombination (HR) for its use in rAAV manufacturing compared to the Tn7-based system. First, we confirmed the functionality of inserted cassette and the absence of undesirable genes into HR-derived baculoviral genomes. Strikingly, we found that the exogenous cassette shown an increased stability over passages when using HR system. Finally, we tested these materials to produce rAAV vectors. The baculoviruses originated from either system lead to high rAAV vector genome yields, with the advantage for the HR method being that the rAAV lots are exempted of undesirable gentamycin and kanamycin genes derived sequences which provides an additional level of safety for the manufacturing of rAAV vectors. Overall, this study highlights the importance of the upstream process and starting biologic materials to generate safer rAAV biotherapeutic products.

Background Patients with severe asthma may present with characteristics representing overlapping phenotypes, making them eligible for more than one class of biologic. Our aim was to describe the profile of severe adult asthma patients eligible for both anti-IgE and anti-IL5/5R and to compare the effectiveness of both classes of treatment in real life. Methods This was a prospective cohort study that included adult severe asthma patients from 22 countries enrolled into the International Severe Asthma registry (ISAR) who were eligible for both anti-IgE and anti-IL5/5R. The effectiveness of anti-IgE and anti-IL5/5R was compared in a 1:1 matched cohort. Exacerbation rate was the primary effectiveness endpoint. Secondary endpoints included long-term-oral corticosteroid (LTOCS) use, asthma-related emergency room (ER) attendance and hospital admissions. Results In the matched analysis (n=350/group), the mean annualized exacerbation rate decreased by 47.1% in the anti-IL5/5R group and 38.7% in the anti-IgE group. Patients treated with anti-IL5/5R were less likely to experience a future exacerbation (adjusted IRR 0.76; 95% CI 0.64, 0.89; p<0.001) and experienced a greater reduction in mean LTOCS dose than those treated with anti-IgE (37.44% vs 20.55% reduction; p=0.023).) There was some evidence to suggest that patients treated with anti-IL5/5R experienced fewer asthma-related hospitalizations (IRR 0.64; 95% CI 0.38, 1.08), but not ER visits (IRR 0.94, 95% CI 0.61, 1.43). Conclusions In real life, both anti-IgE and anti-IL5/5R improve asthma outcomes in patients eligible for both biologic classes, however anti-IL5/5R was superior in terms of reducing asthma exacerbations and LTOCS use.

Zooplankton play a pivotal role in sustaining the majority of marine ecosystems. The distribution patterns and diversity of zooplankton provide key information for understanding the functioning of these ecosystems. Nevertheless, due to the numerous cryptic and sibling species and the lack of diagnostic characteristics for immature developmental stages, the identification of the global-to-local patterns of zooplankton biodiversity and biogeography remains a challenge in different research fields. Here, the spatial and temporal changes in the zooplankton community from the open waters of the southern section of the Gulf of Mexico were assessed using a multilocus sequence analysis and metabarcoding approach based on the genetic information of 18S and cytochrome oxidase c subunit I (COI) genes. Additionally, a multi-scale analysis was implemented to evaluate which environmental predictors may explain the variability in the structure of the zooplankton community. Our finding suggests that the synergistic effects of oxygen, temperature, and longitude (intended as a proxy for still unexplored forces) may explain both spatial and temporal changes in the zooplankton community. Furthermore, the zooplankton distribution likely reflects the coexistence of three heterogeneous ecoregions and a bio-physical partitioning of the studied area. Finally, some taxa were either exclusive or predominant with either 18S or COI data. This may suggest that comprehensive assessments of the zooplankton community may be more accurately met by the use of multi-locus approaches.

Congenital left atrial appendage aneurysm is an extremely rare anomaly. It is seen at any time ranging from fetal to ninth decade of life. Patients with left atrial appendage aneurysm may be asymptomatic, or present with dyspnea, arrhythmias, thromboembolic phenomenon. We report a case of prenatal diagnosis at 26 weeks of gestation , postnatal management and outcome of left atrial appendage aneurysm.

Groundwater plays a crucial role in sustaining global food security but is being over-exploited in many basins of the world. Despite its importance and finite availability, local-scale monitoring of groundwater withdrawals required for sustainable water management practices is not carried out in most countries, including the United States. In this study, we combine publicly available datasets into a machine learning framework for estimating groundwater withdrawals over the state of Arizona. Here we include evapotranspiration, precipitation, crop coefficients, land use, well density, and watershed stress metrics for our predictions. We employ random forests to predict groundwater withdrawals from 2002-2020 at a 2 km spatial resolution using in-situ groundwater withdrawal data available for Arizona Active Management Areas (AMA) and Irrigation Non-Expansion Areas (INA) from 2002-2009 for training and 2010-2020 for validating the model respectively. The results show high training (R2≈ 0.86) and good testing (R2≈ 0.69) scores with normalized mean absolute error (NMAE) ≈ 0.64 and normalized root mean square error (NRMSE) ≈ 2.36 for the AMA/INA region. Using this method, we spatially extrapolate the existing groundwater withdrawal estimates to the entire state and observe the co-occurrence of both groundwater withdrawals and land subsidence in South-Central and Southern Arizona. Our model predicts groundwater withdrawals in regions where production wells are present on agricultural lands and subsidence is observed from Interferometric Synthetic Aperture Radar (InSAR), but withdrawals are not monitored. By performing a comparative analysis over these regions using the predicted groundwater withdrawals and InSAR-based land subsidence estimates, we observe a varying degree of subsidence for similar volumes of withdrawals in different basins. The performance of our model on validation datasets and its favorable comparison with independent water use proxies such as InSAR demonstrate the effectiveness and extensibility of our combined remote sensing and machine learning-based approach.

Objective: Predictors for post-operative reverse remodeling in patients with severe aortic regurgitation (AR) and reduced left ventricular ejection fraction (LVEF) are unknown. We performed low-dose dobutamine stress echocardiography (DSE) in patients with severe AR and reduced LVEF to evaluate the relationship between contractile reserve (CR) and reverse remodeling after surgery. Methods: In 31 patients with chronic severe AR and reduced LVEF (LVEF < 50%), we performed pre-operative DSE, assessed CR and examined whether changes in preoperative DSE were associated with improvement of post-operative LVEF after aortic valve surgery. Results: The pre-operative echocardiographic findings were as follows: left ventricular (LV) end-diastolic dimension: 67 ± 10 mm, LV end-systolic dimension: 52 ± 13 mm and LVEF: 42% ± 8%. All patients underwent aortic valve surgery. Patients with pre-operative LVEF of >45% exhibited a significant increase in LVEF; however, patients with pre-operative LVEF of <45% showed no significant change. When we examined the results of DSE performed in patients with pre-operative LVEF of <45%, ΔLVEF of ≥6% (with CR) during DSE was related to an improvement in post-operative LVEF; ΔLVEF of ≥6% during DSE predicted an improvement in post-operative LVEF, with a sensitivity and specificity of 80% and 85%, respectively. Conclusions: DSE may be a helpful tool for predicting post-operative reverse remodeling in patients with severe AR and moderately reduced LVEF.

With the rapid increase of reported COVID-19 cases, German policymakers announced a 4-week “shutdown light” starting on Nov 2, 2020. Applying mathematical models, possible scenarios for the evolution of the outbreak in Germany are simulated. The results indicate that independent of the effectiveness of the current restrictive measures they might not be sufficient to mitigate the outbreak. Repeated shutdown periods or permanently applied measures over the winter could be successful alternatives.

Nitrogen-fixing Nepalese alder (Alnus nepalensis D. Don.) is a fast-growing early successional species which often forms pure stands in areas affected by landslides and sometimes it occurs mixed with other species in the central Himalayas. In this study, we assessed the distribution of ecosystem carbon storage in plants and soil in a chronosequence of A. nepalensis forest stands in central Himalaya. We examined six forest stands: (1) A. nepalensis-early regeneration (AER) forest, (2) A. nepalensis-late regeneration (ALR) forest, (3) A. nepalensis- mature oak mixed (AMOM) forest, sis tree biomass in different stand AER, APDF, ALR, AMOM, and AMR, AMOO, was 8.97, 51.41, 16.07, 53.74, 144.77, and 101.14 Mg ha-1, respectively. Soil organic C (SOC) in different soil depths in successional stages AER (0-10 cm), APDF (0-30 cm), ALR (0-100 cm), AMOM (0-100 cm), AMR (0-100 cm), and AMOO (0-100 cm) was 3.31, 31.21, 75.47, 157.04, 159.43 and 210.13 Mg ha-1, respectively, with decrease in SOC concentration with increasing soil depth. The ecosystem C storage averaged 15.85, 183, 216.26, 390.32, 403.66, and 500.08 Mg ha-1 in AER, APDF, ALR, AMOM, AMR, and AMOO sites, respectively. Overall, in A. nepalensis forest development markedly ameliorated both vegetation and soil succession in central Himalaya.

In recent years, environmental DNA (eDNA) has received attention from biologists due to its sensitivity, convenience, labor and material efficiency, and lack of damage to organisms. The extensive application of eDNA has opened avenues for the monitoring and biodiversity assessment of amphibians, which are frequently small and difficult to observe in the field, in areas such as biodiversity survey assessment and detection of specific, rare and endangered, or alien invasive species. However, the accuracy of eDNA can be influenced by factors such as ambient temperature, pH, and false positives or false negatives, which makes eDNA an adjunctive tool rather than a replacement for traditional surveys. This review provides a concise overview of the eDNA method and its workflow, summarizes the differences between applying eDNA for detecting amphibians and other organisms, reviews the research progress in eDNA technology for amphibian monitoring, identifies factors influencing detection efficiency, and discusses the challenges and prospects of eDNA. It aims to serve as a reference for future research on the application of eDNA in amphibian detection.

Background: Epitope mapping is an increasingly important aspect of biotherapeutic and vaccine development. Recent advances in therapeutic antibody design and production has enabled candidate mAbs to be identified at a rapidly increasing rate resulting in a significant bottleneck in the characterization of ‘structural’ epitopes, that are challenging to determine using existing high throughput epitope mapping tools. Here, Hydrogen/Deuterium Exchange Mass Spectrometry (HDX-MS) epitope screening workflow was introduced that is well suited for accelerated characterization of epitopes with a common antigen. Main methods and major results: The method is demonstrated on set of 6 candidate mAbs targeting Pertactin (PRN). Using this approach, five of the six epitopes was unambiguously determined using two HDX mixing timepoints in 24 hours total run time, corresponding to substantial decrease in the instrument time required to map a single epitope using conventional HDX workflows. Conclusion: An accelerated HDX-MS epitope screening workflow was developed. The two-timepoint ‘screening’ workflow mapped all six mAbs and generated high confidence epitopes for five of the six mAbs assayed. The substantial improvement in the rate of data collection can advance HDX-MS for higher throughput investigations supporting the ability to evaluate a broader number of mAb candidates at an earlier stage of vaccine development.

Ricinodendron heudelotii, Vitex doniana and Cleome gynandra are three indigenous species of Benin/West Africa. This review focuses on the physicochemical characteristics of their seeds and the nutritional and functional properties of their oils. In this systematic review, scientific articles and reports were used to collect information. The minima, maxima and mean values were considered and converted into a dry basis and/or the same units by using the international system of units when needed to allow comparison. Seeds of Ricinodendron, Vitex and Cleome fat contents were 51.83 g/100 g dw, 28.55 g/100 g dw and 27.35 g/100 g dw, respectively. The three seed oils contained 58.54 to 87% mono and polyunsaturated fatty acids. Ricinodendron seed oil contained a conjugated polyunsaturated fatty acid α-eleostearic acid (49.3–51.1%). Data varied from one author to another due to the methods used. R. heudelotii seeds oil is traditionally used for the treatment of several diseases. Its oil showed phytochemical and antimicrobial properties suggesting its possible use in pharmaceutical industries. Ricinodendron and Vitex seed oil should not be used for cooking at high temperatures or frying because of their high and medium levels of unsaturation.

The evolutionary origins and hybridization patterns of Canis species in North America has been hotly debated for the past 30 years. Disentangling ancestry and timing of hybridization in Great Lakes wolves, eastern Canadian wolves, red wolves, and eastern coyotes is most often partitioned into a 2-species model that assigns all ancestry to grey wolves and/or coyotes, and a 3-species model that includes a third, North American evolved eastern wolf genome. The proposed models address recent or sometimes late Holocene hybridization events but have largely ignored Pleistocene era opportunities for hybridization that may have impacted the current mixed genomes in eastern Canada and the United States. Here, we re-analyze contemporary and ancient mitochondrial DNA genomes with Bayesian phylogenetic analyses to more accurately estimate divergence dates among lineages. We combine that with a review of the literature on Late Pleistocene Canis distributions to illuminate opportunities for ancient hybridization events between extinct Beringian grey wolves (C. lupus) and extinct large wolf-like coyotes (C. latrans orcutti) that we propose as a potentially unrecognized source of introgressed genomic variation within contemporary Canis genomes. These events speak to the potential origins of contemporary genomes and provide a new perspective on Canis ancestry, but do not influence/negate current conservation priorities of dwindling wolf populations with unique genomic signatures and key ecologically critical roles.

G protein-coupled receptors (GPCRs) are a large family of cell surface receptors that play a critical role in nervous system function by transmitting signals between cells and their environment. They are involved in many, if not all, nervous system processes, and their dysfunction has been linked to various neurological disorders representing important drug targets. In this review, we will first discuss the role of the nervous system GPCRs in the modulation of tripartite synapse function and how GPCRs control energy metabolism in the brain. We will then discuss the (patho)physiology and pharmacology of opioid, cannabinoid, acetylcholine, chemokine, and melatonin GPCRs in the nervous system. Furthermore, we will briefly report on adhesion GPCR function in nervous tissues. Finally, we will address orphan GPCRs, their implication in the nervous system function and disease, and the challenges that need to be addressed in the future to deorphanize them.

The evaluation of the lung by ultrasound is an adjunct tool to the clinical assessment. Among different hallmarks at lung ultrasound, B-lines are well known artifacts which are not correlated to identifiable structures but can be used as an instrument for pathological classification. Multiple B-lines are the sonographic sign of lung interstitial syndrome with a direct correlation between the number of B-lines and the severity of the interstitial involvement of lung disease. In neonatology and paediatrics, the quantitative assessment of B-lines is questionable as opposed to in adult medical care. Counting B-lines is an attempt to enrich the clinical assessment and clinical information, and not simply arrive at a dichotomous answer. A semiquantitative or quantitative B-lines assessment was shown to correlate with fluid overload and demonstrated prognostic implications in specific neonatal and paediatric conditions. In neonatology, the count of B-lines is used to predict the need for admission in neonatal intensive care unit and the need for exogenous surfactant treatment. In paediatrics, the B-lines count has the role of quantifying hypervolemia in infants and children receiving dialysis. B-lines as predictors of length of stay in the paediatric intensive care unit after cardiac surgery, as a marker of disease severity in bronchiolitis, or as an indicator of lung involvement from SARS-CoV-2 infection are speculative and not yet supported by solid evidence. Lung ultrasound with the quantitative B-lines assessment is promising. The current evidence allows to use the quantification of B-lines in a limited number of neonatal and paediatric diseases.

Objectives: Clinical studies have shown low toxicity and a favorable safety profile for sirolimus in vascular anomalies. Here, we describe severe adverse events (SAEs) observed during “off-label use”. Methods: We performed a retrospective, multicenter chart review for SAEs during “off-label” sirolimus therapy for vascular anomalies and analyzed these cases by a pre-designed workflow. Results: We identified 17 SAEs in 14 patients diagnosed with generalized lymphatic anomaly (n=4), Gorham-Stout disease (n=2), central conducting lymphatic anomaly (n=1), lymphatic malformation (n=4), tufted angioma (n=1), kaposiform hemangioendothelioma (n=1), and venous malformation in a CLOVES syndrome (n=1). Three patients presented two SAEs. The age at initiation of sirolimus therapy was under the age of 2 years (5x), 2 to 6 years (5x) and older than 12 years (4x). SAEs occurred during the first 3 months of sirolimus therapy (7x), between 3 and 12 months (7x) and after one year of therapy (3x). The most frequent SAE was viral pneumonia (8x) resulting in death due to a metapneumovirus infection in a 3 months old and generalized adenovirus infection in a 28 months old child. Sirolimus blood level at the time of SAEs ranged between 2.7 and 21 ng/L. Five patients were on antibiotic prophylaxis during sirolimus therapy. Conclusions: Most SAEs are observed in the first year of sirolimus therapy; however, SAEs can also occur after a longer treatment period. SAEs are potentially life threatening, especially in early infancy. Presence risk factors, i.e. underlying vascular anomaly or immune status, may contribute to the risk of SAEs.

1. We investigated responses of tomato (Solanum lycopersicum) to belowground presence of two functional guilds of nematodes - plant parasite (Meloidogyne javanica) and entomopathogens (Heterorhabditis bacteriophora, Steinernema feltiae, and S. carpocapsae) - as well as a leaf mining insect (Tuta absoluta) aboveground. Our results indicate that entomopathogenic nematodes (EPNs): 1) induced plant defense responses, 2) reduced root knot nematode (RKN) infestation belowground and 3) reduced herbivore (T. absoluta) host preference and performance aboveground. 2. Concurrently, we investigated the plant signaling mechanisms underlying these interactions using biochemical and transcriptome analyses. We found that both entomopathogen and parasite triggered immune responses in plant roots with shared gene expression. Tomato plants responded similarly to presence of RKN or EPN in the rootzone, by rapidly activating polyphenol oxidase (PPO) and guaiacol peroxidase (GP) activity in roots, but simultaneously suppressed this activity in aboveground tissues.  3. We quantified changes in expression of candidate resistance genes in tomato that may play essential roles in defense response to RKN, which were also coincidentally triggered with EPN. For example, PR-14 expression was greater in plants inoculated with EPN than in plants co-inoculated with both nematode functional guilds. Overall, EPN inoculation directly mediated enhanced plant defense and reduced subsequent RKN infection. Likewise, EPNs may modulate plant defense against RKN invasion, in part, by suppressing active expression of antioxidant enzymes. 4. Inoculation of tomato roots with EPNs belowground reduced both host preference and performance of the aboveground herbivore, T. absoluta.  Inoculations of roots with EPN also triggered an immune response in tomato via up-regulated phenylpropanoid metabolism and synthesis of protease inhibitors (PIs) in plant tissues, which could explain an observed decrease in egg laying and developmental performance exhibited by herbivores on EPN-inoculated plants. 5. Synthesis. Our results add to a growing body of evidence indicating that subterranean EPNs activate systemic acquired resistance (SAR) and/or induced systemic resistance (ISR) in plants with concomitant antagonistic effects on temporally co-occurring subterranean plant pathogenic nematodes and terrestrial herbivores.