Population genomics is a useful tool in the integrated pest management toolbox for elucidating population dynamics, demography, and histories of invasion. However, next-generation sequencing approaches can be hampered by low DNA input from small organisms, such as insect pests. Here, we use a restriction-site associated DNA sequencing approach combined with whole-genome amplification to assess genomic population structure of a newly described pest of canola, the diminutive canola flower midge, Contarinia brassicola. We find that whole-genome amplification prior to library preparation caused a reduction in the overall number of loci sequenced and an increase in overall sequencing depth but had no discernable impact on genotyping consistency for population genetic analysis. Clustering analyses recovered little geographic structure across the main canola production region, but differentiated several geographically disparate populations at edges of the agricultural zone. Given a lack of alternative hypotheses for this pattern, we suggest these data support alternative hosts for this species and thus our canola-centric view of this midge as a pest has limited our understanding of its biology. These results speak to the need for increased surveying effort across multiple habitats and other potential hosts within Brassicaceae, to elucidate both our ecological and evolutionary knowledge of this species as well as potential management implications.

Transient gene expression (TGE) using mammalian cells is an extensively used technology for production of antibodies and recombinant proteins and has been widely adapted by both academic and industrial labs. Chinese Hamster Ovary (CHO) cells have become one of the major work horses for TGE of recombinant antibodies due to their attractive features: post-translational modifications, adaptation to high cell densities, and use of serum-free media. In this study, we describe the optimization of parameters for TGE for antibodies from CHO cells. Through a matrix evaluation of multiple factors including inoculum, transfection conditions, amount and type of DNA used and post-transfection culture conditions, we arrived at an optimized process with higher titer and reduced costs and time, thus increasing the overall efficiency of early antibody material supply. We investigated the amount of coding DNA and the influence of size of the transfection complex on the in vitro efficiency of the transfection. Generation of the transfection complex in serum-free medium leads to the prompt formation of an optimal-sized polyplex, and is independent of the relative amount of coding DNA used for a successful transfection outcome.

Dear Editor,We read with great interest, the article titled “Evaluation of treatments for Bartholin’s cyst or abscess: A systematic review” by authors BJG Illingworth K Stocking M Showell E Kirk JMN Duffy Published in BJOG volume 127, issue 6, May 2020. The article was particularly relevant owing to the fact that Bartholin’s Abscess is a common gynaecological presentation which we encounter regularly in clinical practice.Whilst this article provided some insight into the various techniques for managing Bartholin’s Abscess around the world[1] , there were a few points that we would like to raise. It was not unexpected that the meta-analysis finally concluded that no single technique was superior to the others. This may very well have been due to the different criteria used by these studies to assess the effectiveness of the procedure. Since the failure or success of any technique is largely based on the effect on the quality of life and perception of the outcome by the patient, it may have been more prudent to compare different surgical interventions in terms of patient acceptance, overall satisfaction and long term outcome. Also this analysis compiled data from countries where health services are provided by the private and /or the government sector where the financial incentives for performing procedures vary widely [1] . The article does not elaborate on the proportion of patients who had received pharmacological treatment for varying durations prior to these surgical interventions, which undoubtedly may have influenced the outcome[1]. The analysis also does not include any study where marsupialization was performed under local anaesthetic which may be equally if not more effective than the word catheter at equal cost. In the light of current pandemic situation that the whole world is facing, expertise into minimally more invasive gynaecological procedures to be performed in office setting would also be a key point of consideration. We look forward to the authors comments on these factors.Piyushi Sharma,1 Thangamma Katimada-Annaiah,1 Montasser Mahran,1 Dilip Patil,1 Elvyna Lim,1 Joseph Nattey,1 Tarley Davies1Bedford Hospital NHS TrustReferences1. Illingworth BJG, Stocking K, Showell M, Kirk E, Duffy JMN. Evaluation of treatments for Batholin’s Cyst or abscess: A systematic Review. BJOG 2020;127:671-678

Thrombosis contributes to one in four deaths worldwide and is the cause of a large proportion of mortality and morbidity. A reliable and rapid diagnosis of thrombosis will allow for immediate therapy, thereby providing significant benefits to patients. Molecular imaging is a fast-growing and captivating area of research, in both preclinical and clinical applications. Major advances have been achieved by improvements in three central areas of molecular imaging: 1) Better markers for diseases, with increased sensitivity and selectivity; 2) Optimised contrast agents with improved signal to noise ratio; 3) Progress in scanner technologies with higher sensitivity and resolution. Clinically available imaging modalities used for molecular imaging include, magnetic resonance imaging (MRI), X-ray computed tomography (CT), ultrasound, as well as nuclear imaging, such as positron emission tomography (PET) and single photon emission computed tomography (SPECT). In the preclinical imaging field, optical (fluorescence and bioluminescent) molecular imaging has provided new mechanistic insights in the pathology of thrombembolic diseases. Overall, the advances in molecular imaging, driven by the collaboration of various scientific disciplines, have substantially contributed to an improved understanding of thrombotic disease, and raises the exciting prospect of earlier diagnosis and individualised therapy for cardiovascular diseases. As such, these advances hold significant promise to be translated to clinical practice and ultimately to reduce mortality and morbidity in patients with thromboembolic diseases.

The isosteric heat of adsorption is an important thermodynamic property used to characterise and optimise adsorption processes. In this work, analytic expressions for isosteric heats of adsorption are derived for a collection of commonly used isotherm models and a two-dimensional molecular equation of state based on the SAFT-VR approach. The use of these expressions is presented with an example of adsorption of nitrous oxide, N₂O, on biochar, which is a waste biomass charcoal that exhibits high adsorption potential. The results show that accurate fitting of the adsorption isotherms leads to consistent results obtained with different approaches, however, the predicted isosteric heat of adsorption exhibits strong variations in the regions where experimental data is insufficient such in the region of low pressure/low coverage. Convergence on the prediction of the isosteric heat of adsorption by the different models is only observed in the region where no extrapolation of experimental data is needed.

Most herbivorous insects are diet specialists in spite of the apparent advantages of being a generalist. This conundrum might be explained by fitness trade-offs on alternative host plants, yet evidence of such trade-offs has been elusive. Another hypothesis is that specialization is non-adaptive, evolving through neutral population genetic processes and within the bounds of historical constraints. Here we report on a striking lack of evidence for the adaptiveness of specificity in tropical canopy communities of armored scale insects. We show that specialists abound and that host-use is phylogenetically conservative, but in comparison to generalists, specialists occur on fewer of their potential hosts, and are no more abundant where they do occur. Of course local communities might not reflect regional diversity patterns. But based on our samples, comprising hundreds of species of hosts and armored scale insects at two widely separated sites, host-use specialists do not appear to outperform generalists.

Objective: To review a case of acute urticaria, and to explore the links this presentation shares with hyperthyroidism and anti-thyroid medications, along with management strategies for patients presenting with new-onset urticaria and pre-existing thyroid disease. Design: A retrospective case report on a 23-year-old female with hyperthyroidism, on carbimazole therapy.

Tracing and isolation of close contacts is used to control outbreaks of coronavirus disease 2019 (COVID-19) in China. However, risk factors associated with occurrence of COVID-19 among close contacts have not been well described. 106 household contacts were included in this study, of whom 19 were developed into COVID-19 cases and the secondary attack rate was 17.9%. Multivariable analysis showed increasing risk of occurrence of COVID-19 among household contacts associated with female of index patients (Adjusted Hazard Ratio [aHR] = 5.61, 95%CI = 1.51-20.83), critical disease of index patients ([aHR] = 7.58, 95%CI = 1.66-34.66), effective contact duration with index patients >2 days ([aHR] = 4.21, 95%CI = 1.29-13.73), and effective contact duration >11 days ([aHR] = 17.88, 95%CI = 3.26-98.01).The sex and disease severity of index COVID-19 patients, and longer effective contact duration with COVID-19 confirmed cases could help epidemiologists to identify potential COVID-19 case among household contacts at an early stage.

The SARS-CoV-2 strain of the coronavirus is responsible for the current COVID-19 pandemic, with an ongoing toll of over 5 million infections and 333 thousand deaths worldwide within the first 5 months. Insight into the phylodynamics and mutation variants of this strain is vital to understanding the nature of its spread in different climate conditions. The incidence rate of COVID-19 is increasing at an alarming pace within subtropical Southeast Asian nations with high temperatures and humidity. To understand this spread, we analyzed 60 genome sequences of SARS-CoV-2 available in GISAID platform from 6 Southeast Asian countries. Multiple sequence alignments and maximum likelihood phylogenetic analyses were performed to analyze and characterize the non-synonymous mutant variants circulating in this region. Global mutation distribution analysis showed that the majority of the mutations found in this region are also prevalent in Europe and North America, and the concurrent presence of these mutations at a high frequency in Australia and Saudi Arabia indicate possible transmission routes. Unique spike protein and non-structural protein mutations were observed circulating within a localized area. We divided the circulating viral strains into 4 major groups and 2 sub-groups on the basis of the most frequent non-synonymous mutations. Strains with a unique set of 4 co-evolving mutations were found to be circulating at a high frequency within India, specifically, group 2 strains characterized by two co-evolving NS mutants which alter in RdRp (P323L) and spike protein (D614G) common in Europe and North America. These European and North American variants (Nextstrain clade A2) have rapidly emerged as dominant strains within Southeast Asia, increasing from a 0% presence in January to an 85% presence by May 2020. These variants may have an evolutionary advantage over their ancestral types and could present the largest threat to Southeast Asia for the coming winter.

Intracellular levels of Mg2+ are tightly regulated, as Mg2+ deficiency or excess affects normal plant growth and development. In Arabidopsis, we determined that phospholipase Dα1 (PLDα1) is involved in the stress response to high-magnesium conditions. The T-DNA insertion mutant pldα1 is hypersensitive to increased concentrations of magnesium, exhibiting reduced primary root length and fresh weight. PLDα1 activity increases rapidly after high-Mg2+ treatment, and this increase was found to be dose-dependent. Two lines harboring mutations in the HKD motif, which is essential for PLDα1 activity, displayed the same high-Mg2+ hypersensitivity of pldα1 plants. Moreover, we show that high concentrations of Mg2+ disrupt K+ homeostasis, and that transcription of K+ homeostasis-related genes CIPK9 and HAK5 is impaired in pldα1. Additionally, we found that the akt1, hak5 double mutant is hypersensitive to high-Mg2+. We conclude that in Arabidopsis, the enzyme activity of PLDα1 is vital in the response to high-Mg2+ conditions, and that PLDα1 mediates this response partially through regulation of K+ homeostasis.

This work addresses the design of distributed model predictive control (DMPC) systems for nonlinear processes using machine learning models to predict nonlinear dynamic behavior. Specifically, sequential and iterative distributed model predictive control systems are designed and analyzed with respect to closed-loop stability and performance properties. Extensive open- loop data within a desired operating region are used to develop Long Short-Term Memory (LSTM) recurrent neural network models with a sufficiently small modeling error from the actual nonlinear process model. Subsequently, these LSTM models are utilized in Lyapunov- based DMPC to achieve efficient real-time computation time while ensuring closed-loop state boundedness and convergence to the origin. Using a nonlinear chemical process network exam- ple, the simulation results demonstrate the improved computational efficiency when the process is operated under sequential and iterative DMPCs while the closed-loop performance is very close to the one of a centralized MPC system.

Objectives To assess the diagnostic utility of metagenomic sequencing in pediatric aerodigestive clinic patients being evaluated for chronic aspiration. Clinical metagenomics allows for evaluation of both the respiratory microbiota and the transcriptome of the host immune response. Study Design and Methods Twenty-four children referred to an aerodigestive clinic were enrolled in a prospective, single-site, cross-sectional cohort study. At the time of clinical evaluation under anesthesia, two additional samples were obtained: an upper airway sample and a sample from bronchoalveolar lavage (BAL). Samples were sent for routine culture and analyzed using Explify® Respiratory, a CLIA Laboratory Developed Test (LDT) which identifies respiratory commensals and pathogens through RNA and DNA sequencing. Incidentally obtained host transcriptomics were analyzed to evaluate the host immune response. The results of these studies were correlated with the clinical presentation of the research subjects. Results In ten patients, organisms primarily associated with oral flora were identified in the BAL. Standard culture was negative in three patients where clinical metagenomics led to a result with potential clinical significance. Transcriptomic data correlated with the presence or absence of dysphagia as identified on prior videofluoroscopic evaluation of swallowing. Differentially expressed genes included upregulation of genes involved in the immune response, such as CXCL8 (interleukin 8) and ITGB2 (CD18). Conclusions Clinical metagenomics allows for simultaneous analysis of the microbiota and the host immune response from BAL samples. As the technologies in this field continue to advance, such testing may improve the diagnostic evaluation of patients with suspected chronic aspiration.

OBJECTIVE To describe if weekly determined lung ultrasound (LU) scores in preterm infants born before 32 weeks (PTB32W) change with diuretic therapy. DESIGN This is a secondary analysis of a prospective study about weekly performed LU in PTB32W: we included infants who received diuretics, and compared LU scores according to their evolution on respiratory support (RS) before and after diuretics. RESULTS We included 17 PTB32W. Both groups were similar in terms of median gestational age: 26 weeks (interquartile rank (IQR) 25-28) in the responders group, and 27 weeks (IQR 25-29) in the other; but different in median number of days on invasive mechanical ventilation: 27 (IQR 11-43) vs. 76 (IQR 35-117) in the non-responders group, p=0.03; and number of infants with moderate-severe bronchopulmonary dysplasia: 2 (25%) vs. 8 (89%), p=0.005. The responders group showed lower LU scores at one week after diuretics: median LU score 3 (IQR 0-10) vs. 12 (IQR 12-12) in the non-responders group, p=0.04; and after three weeks of treatment: median value 6 (IQR 3-6) in the responders group vs. 12 (10-15) in the non-responders group, p=0.01. The decrease in RS was achieved since the beginning of diuretics: 4 out of 8 (50%) were extubated in the responders group, and 8 out of 9 (11%) in the non-responders group, p=0.048. CONCLUSIONS There is a group of PTB32W whose LU score improves after diuretics. This change appears later than that observed in the RS.

A growing body of literature has documented myriad effects of human activities on animal behavior, yet the ultimate ecological consequences of these behavioral shifts remain largely uninvestigated. While it is understood that, in the absence of humans, variation in animal behavior can have cascading effects on species interactions, community structure, and ecosystem function, we know little about whether the type or magnitude of human-induced behavioral shifts translate into meaningful ecological change. Here we synthesize empirical literature and theory to create a novel framework for examining the range of behaviorally mediated pathways through which human activities may affect different ecosystem functions. We highlight the few empirical studies that show the potential realization of some of these pathways, but also identify numerous factors that can dampen or prevent ultimate ecosystem consequences. Without a deeper understanding of these pathways, we risk wasting valuable resources on mitigating behavioral effects with little ecological relevance, or conversely mismanaging situations in which behavioral effects do drive ecosystem change. The framework presented here can be used to anticipate the nature and likelihood of ecological outcomes and prioritize management among widespread human-induced behavioral shifts, while also suggesting key priorities for future research linking humans, animal behavior, and ecology.

Title: Age-related risk of household transmission of COVID-19 in SingaporeRachael Pung1, Minah Park2,3, Alex R Cook2,3, Vernon J Lee1,21 Ministry of Health, Singapore2 Saw Swee Hock School of Public Health, National University of Singapore3 National University Health System

To study the hyporheic exchange driven by a single peak flood-induced water level fluctuation (i.e. flood wave), a method combining numerical simulation with theoretical derivation was proposed based on the Inbuk Stream, Korea, where flooding occurs frequently. The hyporheic exchanges induced by different flood waves were investigated by varying amplitude (A), duration (T), wave type parameter (r), and rising duration (tp), which were adopted from the real-time stream stage fluctuations. Additionally, the idea of constant upstream flood volume (CUFV) condition for flood waves was put forward, and the effects of “Botan” (T/A) and peak number (N) on hyporheic exchange were studied. The results showed that the hyporheic exchange flux (q) was controlled by the water level h (sine-type) and its change rate v (cosine-type), and was proportional to the polynomial of them q“∝” (ω∙h+v), where ω is the angular frequency of the flood wave. Based on this mechanism, the influence principles on hyporheic exchanges of the typical flood wave parameters (A, T, r and tp) as well as T/A and N under CUFV condition were clarified. The main characteristic variables of hyporheic exchange, which were maximum aquifer storage and residence time, were positively correlated. They also had positive relations to the integral of the flood wave over time, which increased when the wave became higher, wider, rounder and less skewed. However, when CUFV condition was imposed, the residence time was positively correlated with T/A, whereas the maximum aquifer storage was negatively correlated with T/A. With the increase in N, water exchanged more frequently and some water returned to the stream early, leading to the slight decrease in maximum aquifer storage and residence time. These findings enriched the theory of hyporheic exchange driven by surface water fluctuation and be of great significance to enhance pollutant degradation in the hyporheic zone downstream of reservoirs.

Macrophyte community diversity and composition respond to ecosystem conservation and local environmental factors. In this study, we developed a multidimensional diversity framework for macrophyte communities, including the taxonomic and functional alpha and beta diversity. We used the framework to explore the relationships among water level regimes and these diversity parameters in a case study of China’s Baiyangdian Lake. Analysis of indicators of hydrologic alteration divided the water level from 1959 to 2019 into four regimes (dry, <6.42 m; low, 6.42 to 7.23 m; medium, 7.23 to 8.19 m; high, >8.19 m). Alpha and beta diversity were significantly higher in the medium regime than in the low and high regimes. Redundancy analysis indicated that the maximum water depth significantly affected taxonomic alpha diversity, and total nitrogen (TN) and chemical oxygen demand (COD) concentration significantly affected functional alpha diversity, respectively. Mantel tests showed that TN, Secchi depth, and water depth in the high water level regime significantly increased the total beta diversity and turnover components. TN was the main factor that increased total taxonomic beta diversity. Interspecific competition decreased with the decreasing range (variation) of TN values and differed opposite with the variation of COD values in response to increasing water level, and reached its maximum in the medium regime. Ecosystem stability was promoted by maintaining high species richness and evenness and high differences among communities, and by reducing competition. Based on our results, the water level should be maintained between the medium and high water level regimes to promote restoration of the macrophyte community and improve ecosystem stability.

Background: We report our experience in aortic arch repair with the E-vita Open hybrid prosthesis and describe the changes in our technique over time. Methods: Between October 2013 and December 2019, 56 patients underwent a total aortic arch replacement with the E-vita Open hybrid prosthesis. Main indications were thoracic aorta aneurysm (n=27) and acute type A aortic dissection (n=18). We analyze the technique and results in the overall series, and compare both between our early (Group I, 25 patients) and late experience (Group II, 31 patients). Results: Overall in-hospital mortality was 7.1% (4), and permanent stroke and spinal cord injury were 3.6% and 1.8% respectively. 15 patients (26.8%) underwent a planned second procedure on the distal aorta: 13 endovascular, 1 open and 1 hybrid. Survival at 1 and 3 years was 90.7% and 80.7%. Group II included more patients with acute dissection (45.2% vs 16%, p=0.02), a higher rate of bilateral cerebral perfusion (100% vs 64%, p<0.001), left subclavian artery perfusion during lower body circulatory arrest (87.1% vs 0%, p<0.001), early reperfusion (96.8% vs 40%, p<0.001), and zone 0-2 distal anastomosis (100% vs 72%, p=0.02). In-hospital mortality (3.2% vs 12%) and permanent stroke (0% vs 8%) tended to be lower in Group II. Conclusions: Total arch replacement with E-vita Open hybrid prosthesis in complex thoracic aorta disease is safe. One-stage treatment is feasible when pathology does not extend beyond the proximal descending thoracic aorta. In any case, it facilitates subsequent procedures on distal aorta if needed.