Increased circulating CRTH2+Tregs are associated with asthma control and exacerbation Short running title:Increased circulating CRTH2+Tregs among patients with asthmaTeerapol Chantveerawonga, Sasipa Sangkangjanavanicha,b, Chirawat Chiewchalermsria,c, Panitan Pradubpongsaa, Wat Mitthamsiria, Sarawut Jindaratd, Ming Wange, Mübeccel Akdisf, Milena Sokolowskaf, Cezmi A. Akdisf, Atik Sangasapaviliyaa, Tadech Boonpiyathada,faDivision of Allergy and Clinical Immunology, Department of Medicine, Phramongkutklao Hospital, Bangkok, Thailandb Division of Allergy, Immunology and Rheumatology, Department of Medicine, Faculty of Medicine Ramathibodi Hospital, Mahidol University, Bangkok, ThailandcDepartment of Medicine, Panyananthaphikkhu Chonprathan Medical Center, Srinakharinwirot University, Nonthaburi, ThailanddDepartment of Pharmacology, Phramongkutklao College of Medicine, Bangkok, ThailandeDepartment of Otolaryngology, Head and Neck Surgery, Beijing TongRen Hospital, Capital Medical University, and the Beijing Key Laboratory of Nasal Diseases, Beijing Institute of Otolaryngology, Beijing, ChinafSwiss Institute of Allergy and Asthma Research (SIAF), University of Zurich, and the Christine Kühne-Center for Allergy Research and Education (CK-CARE), Davos, Switzerland

Unloading compliance (UC) method and normalization method (NM) are two of the most commonly used methods for determining the fracture toughness of materials. However, considerable differences often exist in the fracture toughness determined by these two methods, which solicits a new method to determine the fracture toughness accurately. In this paper, the compliance of crack length differences as measured by the crack length difference ratio Si is discovered, analysed and verified by experiments. Based on this compliance, a new accurate method, known as AJR, is developed and verified by test results. Factors that exhibit the advantages of the developed new AJR method are also investigated. It is found that the J-R curves determined by the new AJR method are more accurate than those determined by UC and NM. The new AJR method should be the first choice for steels with a small strain hardening ratio and low effective yield strength, and thicker CT specimens with shallower initial crack length. This is because the disagreement between UC and NM is unacceptably large. The developed new AJR method and the results presented in this paper can assist engineers and researchers to determine J-R curves and fracture toughness of steels more accurately and can contribute to the body of knowledge of fracture mechanics.

Functional rehabilitation of post-laryngectomy patients is a challenging task; this includes achieving optimal speech and swallowing outcomes. Botox injections are utilised to improve pharyngoesophageal segment tonicity to improve speech and swallowing outcomes. We propose a novel and efficacious technique using a portable video-laryngoscope, GlideScope®, to directly visualise the injection of botulinum toxin in total laryngectomy patients with severe radiation-induced changes.

Synthetic biology has promoted the development of biosensors as tools for detecting trace substances. In the past, biosensors based on synthetic biology have been designed on living cells, but the development of cell biosensors has been greatly limited by defects such as the obstruction of cell membrane. However, the advent of cell-free synthetic biology addresses these limitations. Biosensors based on the cell-free protein synthesis system have the advantages of higher safety, higher sensitivity, and faster response time over cell biosensors, which makes cell-free biosensors have a broader application prospect. This review summarizes the workflow of various cell-free biosensors, including the identification of analytes and signal output. The detection range of cell-free biosensors is greatly enlarged by different recognition mechanisms and output methods. In addition, the review also discusses the applications of cell-free biosensors in environmental monitoring and health diagnosis, as well as existing deficiencies and aspects that should be improved. In the future, through continuous improvement and optimization, the potential of cell-free biosensors will be stimulated, and their application fields will be expanded.

We report a case of a 17-year-old healthy male presenting with multisystem hyperinflammatory shock temporally associated with COVID-19. Cardiac involvement was suspected based on evidence of significant cardiac injury (elevated cardiac biomarkers, electrocardiographic and echocardiographic abnormalities). Cardiac magnetic resonance imaging was performed demonstrating global biventricular systolic dysfunction, as well as a small area of T2 hyperintensity and mid wall late gadolinium enhancement. This case discusses the varied cardiac involvement in pediatric patients with COVID-19 infection and highlights that cardiac injury is not just limited to hyperinflammatory syndrome related global dysfunction but a more focal myocarditis can also be seen.

Plant roots interact with rhizosphere microbes to accelerate soil organic matter (SOM) mineralization and promote nutrient acquisition. Root-mediated changes in SOM turnover largely depend on root-C input and soil nutrient availability. Hence, interspecific competition and nutrient uptake dynamics over plant development stages as well as spatiotemporal variability in C input may modify SOM turnover. To investigate the effect of intraspecific competition on SOM mineralization at three growth stages (heading, flowering and ripening), we grew maize (C4 plant) under three planting densities on a C3 soil. 13C-natural abundance and 15N-pool dilution were applied in situ to determine C- and N-mineralization rates. Soil C- and N-mineralization rates were tightly coupled and peaked at maize flowering. However, the C-to-N-mineralization ratio increased with N, indicating that microbes mineralize N-rich components to mine SOM for N. Furthermore, intraspecific competition did not affect root biomass; instead, plants shaped root morphology towards higher specific root length as an efficient strategy competing for nutrient. Hence, root morphologic traits rather than root biomass per se were positively related to C- and N-mineralization. Overall, plant competition for nutrients controlled the intensity and mechanisms of soil C- and N-turnover by the adaptation of root traits and nutrient depletion.

The purpose of this article is to introduce an application to draw the asymptotes of Bode diagram module and phase from each constituent elementary factors of any transfer function for minimum and non-minimal phase systems without transport delay. The Bode diagram is the most used tool in the frequency response method. Python was used to program the application to perform the operations as well as the Qt5 Design for the simple graphical interface for the application and all this in the Linux operating system. The application purpose is to assist students in learning the concept and drawing of Bode diagram. For students the non-minimum phase system Bode diagram is more difficult to draw than a minimum phase system due to the presence of zeros and/or poles on right half of s-plane. The phase asymptotes of a quadratic factor was closest to the real phase curve around the corresponding undamped natural frequency and this can be observed in the example showed in this article. This example must be used as a help and not a simply to solve a problem.

Objectives To summarise current practices in the diagnosis and management of Deep Neck Space Infections (DNSIs) To inform future studies in developing a framework in the management of DNSIs Design This review was registered on PROSPERO (CRD42021226449) and reported in line with PRISMA guidelines. All studies from 2000 that reported the investigation or management of DNSI were included. The search was limited to English language only. Databases searched included AMED, Embase, Medline and HMIC. Quantitative analysis was undertaken with descriptive statistics and frequency synthesis with 2 independent reviewers. A qualitative narrative synthesis was conducted using a thematic analysis approach. Setting Secondary or Tertiary Care centres that undertook management of Deep Neck Space Infections. Participants All adult patients with a deep neck space infection. Main outcome measures The role of imaging, radiologically guided aspiration and surgical drainage in DNSIs. Results 60 studies were reviewed. 31 studies reported on imaging modality, 51 studies reported treatment modality. Aside from a single RCT all other studies were observational (n=25) or case series (n=36). CT was used to diagnose DNSI in 78% of patients, Mean percentage of management with open surgical drainage was 81% and 29.4% for radiologically guided aspiration. Qualitative analysis identified 7 major themes were identified on DNSI. Conclusion There are limited methodologically rigorous studies investigating DNSIs. CT imaging was the most used imaging modality. Surgical drainage was commonest treatment choice. Areas of further research on epidemiology, reporting guidelines and management are required.

Background: The COVID19 pandemic gripped every nation’s healthcare system and provisions on all levels. In cardiac and aortic surgery, as it is with most specialities, elective surgeries were halted. Aims of the study: We captured reflections, contingencies, and current practices across of high-volume centres in cardiac and aortic surgery globally. We also aimed this study to assess decision on prioritization of the surgical patients, the need for personal protection equipment and choice of preoperative investigations in current dynamic and fluid climate. Methods: A validated web-based questionnaire was constructed and was circulated to the international surgeons amongst high volume cardiac and aortic surgery centres. Their intrinsic feedback on decision making, impact of the lockdown and perspectives for the future ahead us all were noted. Mixed method approach was constructed. Qualitative data analysis was introduced to signify the impact globally. Results: Overall, 23 centers from 18 countries participated in this international study. 91.7% of the respondents stopped operating on elective patients during the pandemic. Majority of the surgeons agreed that acute aortic dissection (87.1%) should be operated as emergency procedure and stable triple vessel disease (87.1%) to be considered as elective procedure. Three-fifth (60%) of the respondents relied on CT chest as a preoperative screening modality. Conclusion: In the present climate where there is paucity of evidence, this will give an interim consensus for the cardiac surgeons. With the increase in cumulative number of COVID19 patients, careful utilization of the resources regarding hospital beds and manpower is of paramount importance.

SARS-CoV2 infection has caused an increase in mortality and morbidity but with vaccination, the disease severity has significantly reduced. But with emergence of various variants of concerns (VOCs), the vaccine breakthrough infection has also increased. Here, we studied circulating spike specific T follicular response (cTfh) in infection naïve vaccinees and convalescent vaccinees (individuals who got delta breakthrough infection after 2 doses of BBV152 vaccine) to understand their response as they are the most crucial cells that are involve in vaccine mediated protection by helping in B-cell maturation. Our results indicated that cTfh cells in both the groups recognized wild type and delta spike protein but memory response to wild type spike was superior in infection naïve than in convalescent group. The cytokine response particularly IL-21 from cTfh cells was also higher in infection naïve than convalescent vaccines indicating a dampened cTfh response in convalescent vaccines after breakthrough infection. Also, there was positive correlation between IL-21 from cTfh cells and neutralizing antibodies of infection naïve vaccinees. Multiple cytokine analysis also revealed higher inflammation in convalescent vaccinees. Our data indicated necessity of third booster dose may be individual specific depending in the steady state functional phenotype of immune cells.

Objectives: A significant proportion of COVID-19 patients may have cardiac involvement including arrhythmias. Although arrhythmia characterization and possible predictors were previously reported, there are conflicting data regarding the exact prevalence of arrhythmias. Clinically applicable algorithms to classify COVID patients’ arrhythmic risk are still lacking, and are the aim of our study. Methods: We describe a single center cohort of hospitalized patients with a positive nasopharyngeal swab for COVID-19 during the initial Israeli outbreak between 1/2/2020 –30/5/2020. The study’s outcome was any documented arrhythmia during hospitalization, based on daily physical examination, routine ECG’s, periodic 24-hour Holter, and continuous monitoring. Multivariate analysis was used to find predictors for new arrhythmias and create classification trees for discriminating patients with high and low arrhythmic risk. Results: Out of 390 COVID-19 patients included, 28 (7.2%) had documented arrhythmias during hospitalization, including: 23 atrial tachyarrhythmias, combined atrial fibrillation (AF) and ventricular fibrillation, ventricular tachycardia storm, and 3 bradyarrhythmias. Only 7/28 patients had previous arrhythmias. Our study showed significant correlation between disease severity and arrhythmia prevalence (p<0.001) with a low arrhythmic prevalence among mild disease patients (2%). Multivariate analysis revealed background heart failure (CHF) and disease severity are independently associated with overall arrhythmia while age, CHF, disease severity, and arrhythmic symptoms are associated with tachyarrhythmias. A novel decision tree using age, disease severity, CHF, and troponin levels was created to stratify patients into high and low risk for developing arrhythmia. Conclusions: Dominant arrhythmia among COVID-19 patients is AF. Arrhythmia prevalence is dependent on age, disease severity, CHF, and troponin levels. A novel simple Classification tree, based on these parameters, can discriminate between high and low arrhythmic risk patients.

Aims: Mirvetuximab soravtansine is the first-in-class antibody-drug conjugate approved in November 2022 for the treatment of folate receptor-α positive ovarian cancer. The aim of this study was to develop a population pharmacokinetic (PK) model to describe the concentration-time profiles of mirvetuximab soravtansine, the payload (DM4) and a metabolite (S-methyl-DM4). Methods: Mirvetuximab soravtansine was administered intravenously from 0.15 to 7 mg/kg to 543 patients with predominantly platinum-resistant ovarian cancer in three clinical studies, and the plasma drug concentrations were analyzed using a nonlinear mixed-effects modelling approach to estimate the PK parameters, inter-individual variabilities, and residual errors. Stepwise covariate modelling was performed to identify covariates. Results: We developed a semi-mechanistic population PK model that included linear and nonlinear routes for the elimination of mirvetuximab soravtansine and a target compartment for the formation and disposition of the payload and metabolite in tumor cells. The model adequately described the concentration-time profiles for the three analytes. Patient body weight, serum albumin, and age were identified as the major covariates. Exposures in patients with renal or hepatic impairment were estimated. The effect of inhibition of cytochrome P450 (CYP) 3A4 on drug exposures was also evaluated. Conclusions: There is no need for dose adjustment due to covariate effects for mirvetuximab soravtansine administered at the recommended dose of 6 mg/kg based on adjusted ideal body weight. The model also showed that dose adjustment is not required for patients with mild or moderate renal impairment, mild hepatic impairment, or when concomitant weak and moderate CYP3A4 inhibitors are used.

Background: Although diastolic dysfunction is common among patients treated with cancer therapy, no clear evidence has been shown that it predicts systolic dysfunction. This study evaluated the correlation of longitudinal diastolic strain time (Dst) with the routine echocardiography diastolic parameters and to estimated its role in the early detection of cardiotoxicity among patients with active breast cancer. Methods: Data were collected as part of the Israel Cardio-Oncology Registry (ICOR), a prospective registry enrolling all adult patients referred to the cardio-oncology clinic. All patients with breast cancer, planned for Doxorubicin therapy were included. Echocardiography, including Global longitudinal systolic strain (GLS) and Dst, was assessed at baseline before chemotherapy (T1), during Doxorubicin therapy (T2) and after the completion of Doxorubicin therapy (T3). Cardiotoxicity were determined by GLS relative reduction of ≥15%. Dst was assessed as the time measured (ms) of the myocardium lengthening during diastole. =diastolic time (ms) measured. Results: Among 69 patients, 67 (97.1%) were females with a mean age 52±13years. Diastolic strain timeDst measurement was significantly associated with the standard routine diastolic parameters. Significant GLS reduction was observed in 10 (20%) patients at T3 . Both in a univariate and a multivariate analyses the change in Ds basal time from T1 to T2 emerged to be significantly associated with GLS reduction at T3 (p<0.04). Conclusions: Among breast cancer patients, Dst time showed high correlation to standard the routine diastolic echocardiography parameters. Relative reductionChange in Ds basal time emerged associated with clinically significant systolic dysfunction as measured by GLS reduction.

Background A dose of 5mg/kg lidocaine is considered appropriate for paediatric airway topicalisation. Existing literature suggests younger children are susceptible to toxic lidocaine plasma levels and achieve this at a faster rate. Aims The primary outcome of this study was to ascertain peak plasma lidocaine levels after topicalisation for airway endoscopy. Secondary endpoints included: time to peak lidocaine plasma levels, signs of lidocaine toxicity (restricted to ECG changes or seizures when under anaesthesia) and clinical adverse events of laryngospasm, coughing or desaturation during the procedure. Methods Data was collected prospectively over 18 months at Royal Manchester Children’s Hospital. Children aged 0-8 years undergoing elective diagnostic or therapeutic airway endoscopy were included within the study. Standardised 2% lidocaine was used for airway topicalisation. Dose varied depending upon practitioner usual practice. Venous blood sampling occurred at 5, 10, 15 and 20 minutes post administration and plasma lidocaine levels (ng/ml) analysed. Results A significant relationship exists between higher peak plasma levels and ages <18 months (p=0.00973). Strong linear correlation exists between weight and age for our cohort (r=0.88). Higher peak plasma lidocaine levels occur with total dose volumes between 2 and 3mls of 2% lidocaine local anaesthetic (p=0.03) compared with <2ml total dose volumes. Data suggests a potential relationship of lower weights achieving higher peak plasma levels (p=0.0516). Reduced IQR variation of peak plasma lidocaine levels exists when lidocaine dosing is <5mg/kg. Conclusions Age and total dose volume of topicalised lidocaine have a significant relationship with plasma lidocaine levels. A dose of 5mg/kg topicalised lidocaine for paediatric airway endoscopy is safe and provides good operating conditions. Lower patient weights trend toward higher peak lidocaine plasma concentrations and require further investigation.

Since a polyvalent strategy has recently been assumed to be adopted by Deinococcus radiodurans that can generate various resistance against many different detrimental sources of oxidative damage (e.g. reactive oxygen species, heavy metal ions and ionising radiation), investigating more than one restorative metabolic activities and their interrelation of the very same entities of Deinococcus radiodurans is of great significance for exploring its polyextremophile nature, which will be insightful for obtaining fundamental generic insights into life sustainability. Herein, we apply mainly fluorescence microscopy and back reflection microscopy to visibly assess the respective activities of superoxide radical generation and silver ion metabolism for individual Deinococcus radiodurans. Strikingly, only a minority (<20%) of the bacteria which show low superoxide radical levels is revealed to exhibit considerable formation of silver nanoparticles whilst those containing more superoxide radicals all show minimum silver ion metabolism. The discovery of the strong negative correlation for the small subpopulation between the two visualised different metabolic activities not only provides direct experimental evidence in terms of bacterial functionality for the inferred survival regime of the extreme microbe, but also suggests a new way of chemically examining biology from the perspective of inter-functional relationship.

Global emergence and re-emergence of Porcine epidemic diarrhea virus (PEDV), an Alphacoronavirus which causes a highly contagious enteric disease, have led to several studies addressing its variability. The aim of this study was to characterize the infection of weaned pigs with Swine enteric coronavirus (SeCoV) -a chimeric virus most likely originated from a recombination event between PEDV and Transmissible gastroenteritis virus, or its mutant Porcine respiratory coronavirus-, and two PEDV G1b variants, including a recently described recombinant PEDV-SeCoV (rPEDV-SeCoV), as well as to determine the degree of cross-protection achieved against the rPEDV-SeCoV. For this purpose, forty-eight 4-week-old weaned pigs were randomly allocated into four groups of 12 animals; piglets in groups B, C and D were orally inoculated with a PEDV variant (B and D) or SeCoV (C), while piglets in group A were mock inoculated and maintained as controls. At day 20 post-infection all groups were exposed to rPEDV-SeCoV; thus, group D was subjected to a homologous re-challenge, groups B and C to a heterologous re-challenge (PEDV/rPEDV-SeCoV and SeCoV/rPEDV-SeCoV, respectively) and group A was primary challenged (-/rPEDV-SeCoV). Clinical signs, viral shedding, microscopic lesions and specific humoral and cellular immune responses (IgG, IgA, neutralizing antibodies and IgA and IFN-γ-secreting cells) were monitored. After primo-infection all three viral strains induced an undistinguishable mild-to-moderate clinical disease with diarrhea as the main sign and villus shortening lesions in the small intestine. In homologous re-challenged pigs, no clinical signs or lesions were observed, and viral shedding was only detected in a single animal. This fact may be explained by the significant high level of rPEDV-SeCoV-specific neutralizing antibodies found in these pigs before the challenge. In contrast, prior exposition to a different PEDV G1b variant or SeCoV only provided partial cross-protection, allowing rPEDV-SeCoV replication and shedding in feces.

The production of recombinant proteins usually reduces cell fitness and the growth rate of producing cells. The growth disadvantage favors faster-growing non-producer mutants. Therefore, continuous bioprocessing is hardly feasible in Escherichia coli due to the high escape rate. We investigated the stability of E. coli expression systems under long-term production conditions and how metabolic load triggered by recombinant gene expression influences the characteristics of mutations. We conducted iterated fed-batch-like microbioreactor cultivations under production conditions. We used the easy-to-produce green fluorescent protein (GFP) and a challenging antigen-binding fragment (Fab) as model proteins, and BL21(DE3) and BL21Q strains as expression hosts. In comparative whole genome sequencing analyses, we identified mutations that allowed cells to grow unhindered despite recombinant protein production. A T7 RNA polymerase expression system is only conditionally suitable for long-term cultivation under production conditions. Mutations leading to non-producers occur in either the T7 RNA polymerase gene or the T7 promoter. The host RNA polymerase-based BL21Q expression system remained stable in the production of GFP in long-term cultivations. For the production of Fab, mutations in lacI of the BL21Q derivatives had positive effects on long-term stability. Our results indicate that adaptive evolution carried out with genome-integrated E. coli expression systems in microtiter cultivations under industrial relevant production conditions is an efficient strain development tool for production hosts.

We represent a case with simultaneous COVID-19 and acute type A aortic dissection. Type A aortic dissection, an aortic catastrophic event, seems to have higher mortality on coexistence with COVID-19.