Allergy related outcomes at 12 months in the CORAL Birth cohort of Irish children born during the first COVID 19 lockdown.Word count: 1248, References: 10

Background and aim of the study Objective of this analysis was to use coronary computed tomography in patients with normal tricuspid aortic valves to perform detailed aortic root and aortic valve geometric analysis with focus on asymmetry of the three leaflets. Materials and methods We analyzed aortic valves in 70 anonymized coronary computed tomography angiograms. Mean patient age was 53 ± 11 years. All aortic valves were tricuspid, without calcifications and aortic roots were of normal dimensions. Asymmetry of the three leaflets in individual patients was assessed by calculating absolute and relative differences between the largest and the smallest of the three leaflets. Results Some degree of asymmetry was present in all analyzed valves. Absolute and relative differences for free margin length were 3.2 ± 1.4 mm and 9.3 ± 3.8%, respectively. The largest relative difference was noted in coaptation area (36.5 ± 16.5%) and the smallest in leaflet effective height (6.1 ± 4.8%). Using predefined cut-off criteria for absolute differences in leaflet dimensions, 86% of the valves were classified as asymmetric. Conclusions Equal free margin length of the three leaflets is not needed for normal tricuspid aortic valve function. Aligning the leaflet free margin length in standardized aortic valve repair may not be necessary in tricuspid aortic valve repair, whereas equalization of effective leaflet heights is.

The osteopontin released from mesenchymal stem cells (MSC) undergoing lineage differentiation can negatively influence the expansion of hematopoietic stem cells (HSCs) in co-culture systems developed for expanding HSCs. Therefore, minimising the amount of osteopontin in the co-culture system is important for the successful ex vivo expansion of HSCs. Towards this goal, a bioengineered 3D-microfibrous matrix that can maintain MSCs in less osteopontin releasing condition has been developed and its influence on the expansion of HSCs has been studied. The newly developed 3D-matrix significantly decreased the release of osteopontin, depending on the MSC culture conditions used during the priming period before HSC seeding. The culture system with the lowest amount of osteopontin facilitated more than 40-fold increase in HSC number in 1 weeks’ time period. Interestingly, the viability of expanded cells and the CD34+ pure population of HSCs found to be the highest in the low osteopontin containing system. Therefore, bioengineered microfibrous 3D-matrices seeded with MSCs, primed under suitable culture conditions can be an improved ex vivo expansion system for HSC culture.

Comment on: Burden of central nervous system complications in sickle cell disease: A systematic review and meta-analysis R. Grant Steen, PhD103 Van Doren PlaceChapel Hill, NC [email protected] Editor;I read with interest a recent paper in Pediatric Blood & Cancer(1), about cognitive impairment in children with sickle cell disease (SCD). I’m always grateful to have my work cited, especially since our paper is almost 20 years old (2).However, it seems that Lee et al . have gone to great lengths to mischaracterize our work. They write (pg. 5) that, “SCI [silent cerebral infarct] and absence of SCI/CVA [cerebrovascular accident] were verified by review of brain MRIs in all studies except Ghafuri et al . and Steen et al . In these studies, absence of SCI/CVA was defined as no mention of cerebral infarcts in the subject’s medical records.” Also on pg. 5, they write that, “An alternative analysis was conducted excluding studies with insufficient definition of overt stroke and/or SCI based on clinical opinion (Ghafuriet al . and Steen et al .)”. I cannot speak for Ghafuriet al , but we did a careful review of MRI findings for every single patient in our study, as is necessary for a paper in theAmerican Journal of Neuroradiology . This should be pellucidly clear because the title of our paper is, “Cognitive Impairment in Children with Hemoglobin SS Sickle Cell Disease: Relationship to MR Imaging Findings and Hematocrit ”. All our SCD patients were evaluated by at least two neuroradiologists, and patients thought to be abnormal were evaluated by a third neuroradiologist blinded as to previous findings (2). One wonders what other errors may be present in Leeet al (1).Fortunately, this mischaracterization of our work did not lead to incorrect conclusions (1). A comparison of Table 1 to Supplementary Table 6, in which our paper and Ghafuri et al were deleted from analysis, shows that none of the major findings were changed much by deleting these papers (1). I agree with most conclusions in Lee et al .; in fact, I would go further. In uncited work from St. Jude Children’s Research Hospital, we found that children with SCD can suffer cognitive impairment even in the absence of MRI evidence of brain injury (3). We tested a hypothesis that children with SCD who are completely normal by MRI can still be cognitively impaired, as predicted by a model of diffuse brain injury. A total of 54 patients with hemoglobin SS (average age 10.9 years ± 2.9 years SD) were examined with the Wechsler Intelligence Scale for Children-III (WISC-III) and were randomly matched by age, race, and gender to healthy children from the Wechsler normative database. Patients were also imaged at 1.5 Tesla with standard imaging sequences. Among 30 patients judged normal by MRI, there were substantial deficits in Wechsler Full-Scale IQ, Verbal IQ, and Performance IQ (all P < 0.01), when compared to African-American controls. The patient Wechsler Full-Scale IQ was 12.9 points lower than that of matched controls and decreased as a function of age (P = 0.014). These findings strongly suggest that there is diffuse brain injury in SCD patients, perhaps associated with chronic hypoxia (3).

Background Cow’s milk protein allergy (CMPA) is one of the most common food allergies in infancy. Most infants with CMPA tolerate baked milk from diagnosis and gradually acquire increased tolerance. Nevertheless, parents often display significant anxiety about this condition and a corresponding reluctance to progress with home introduction of dairy due to concerns about possible allergic reactions. Objective: To evaluate the impact on gradual home introduction of foods containing cows milk after a supervised, single low dose exposure to whole milk at time of diagnosis. Methods Infants less than 12 months old, referred with suspected IgE-mediated cow’s milk allergy were recruited to an open-label randomised, controlled trial of intervention - a single dose of fresh cow’s milk, using the validated dose of milk that would elicit reactions in 5% of CMPA subjects - the ED 05 – vs routine care. Both groups implemented graded exposure to CM (using the 12 step MAP Milk Tolerance Induction Ladder), at Home. Parents completed food allergy quality of life and State and Trait Anxiety Inventories (STAI). Main outcome measures were milk ladder position at 6 months and 12 months post randomisation. Results: Sixty patients were recruited, 57 (95%) were followed to 6 months. By 6 months 27/37 (73%) intervention subjects had reached step 6 or above on the milk ladder compared to 10/20 (50%) control subjects (p=0.048). By 6 months 11/37 (30%) intervention subjects had reached step 12 (ie drinking unheated cow’s milk) compared to 2/20 (10%) of the controls (p=0.049). Twelve months post randomisation 31/36(86%) of the intervention group and 15/19(79%) of the control group were on step 6 or above. However, 24/37 (65%) of the intervention group were at step 12 compared to 7/20 (35%) of the control group (p=0.03). Maternal STAIs were significantly associated with their infants’ progress on the milk ladder and with changes in skin prick test and spIgE levels at 6 and 12 months. Conclusion This study demonstrates the safety and effectiveness of introduction of baked milk implemented immediately after diagnosis of cows milk allergy in a very young cohort. A supervised single dose of milk at the ED 05 significantly accelerates this further, probably by giving parents the confidence to proceed. Maternal anxiety generally reflects infants’ progress towards completion of the milk ladder, but pre-existing high levels of maternal anxiety are associated with poorer progress.

Successful Extubation in Preterm InfantsTo the Editor,I read with great interest the article by O’Connor et al1 describing factors that lower the likelihood of successful extubation by day 14 in mechanically ventilated neonates with or at risk of developing bronchopulmonary dysplasia (BPD), born at <30week gestational age (GA). These factors were younger GA at birth, and at the time of commencing steroids had higher MAPs and had higher oxygen requirements. They studied 287 neonates and noted that each additional week of GA at birth led to a 1.53 increase in the odds of successful extubation. Higher average fraction of inspired oxygen (FiO2) requirements in the preceding 24 h resulted in a 0.94 decrease in the odds of successful extubation and higher mean airway pressure (MAP) resulted in 0.76 decrease in odds of successful extubation.I have been using the probability of successful extubation calculator (http://extubation.net/), which incorporates the same factors as described by Connor et al 1. The calculator was described earlier by Gupta et al 2. They incorporated GA, MAP and FiO2 in the calculation, same as Connor et al 1. The only difference was the inclusion of the respiratory severity score (RSS), which is nothing but a product of MAP and FiO2 [(RSS= mean airway pressure (cm H2O) x FiO2 (0.21-1.00)].Although, we have a probability score model for successful extubation among preterm infants, as described by Gupta et al 2, I do agree with Connor et al 1 that we need to develop prognostic scoring model for intubated preterm infants.References:O’Connor K, Hurst C, Llewellyn S, Davies M. Factors associated with successful extubation following the first course of systemic dexamethasone in ventilator-dependent preterm infants with or at risk of developing bronchopulmonary dysplasia [published online ahead of print, 2022 Jan 7]. Pediatr Pulmonol . 2022;10.1002/ppul.25821. doi:10.1002/ppul.25821Gupta D, Greenberg RG, Sharma A, et al. A predictive model for extubation readiness in extremely preterm infants. J Perinatol . 2019;39(12):1663-1669. doi:10.1038/s41372-019-0475-xAuthor:Shabih Manzar, MDLouisiana State University Health Sciences CenterDepartment of Pediatrics1501 Kings HighwayShreveport, LA 71103Phone: 318-626-1623Fax: 318-675-6059Email: [email protected] contribution: Dr. Manzar wrote the draft.Funding and financial support: NoneConflict of interest: None

anterior mini‐thoracotomy in multivessel coronary revascularization by Cyn I read with interest the article, routine minimally invasive approach via left ak and colleagues1. The main problem of mini anterior thoracotomy coronary revascularization is the limited or even bad exposure to the right coronary and posterior obtuse marginal branches which may affect the quality of the distal anastomosis. This is the most important technical point of a successful revascularization.

Myeloid cell leukemia-1 (MCL1), an anti-apoptotic BCL-2 family protein plays a major role in the control of apoptosis as the regulator of mitochondrial permeability which is deregulated in various solid and hematological malignancies. Interaction of the executioner proteins Bak/Bax with anti-apoptotic MCL1 and its cellular composition determines the apoptotic or survival pathway. This study highlighted the deleterious MCL1-Bax stabilizing effect of the mutation V220F on MCL1 structure through computational protein-protein interaction predictions and molecular dynamics simulations. The single point mutation at V220F was selected as it is residing at the hydrophobic core region of BH3 conserved domain, the site of Bax binding. The molecular dynamics simulation studies showed increase in stability of the mutated MCL1 before and after Bax binding comparable with the native MCL1. The clusters from free energy landscape found out structural variation in folding pattern with additional helix near the BH3 domain in the mutated structure. This loop to helix structural change in the mutated complex favored stable interaction of the complex and also induced Bax conformational change. Moreover, molecular mechanics based binding free energy calculations confirmed increased affinity of Bax towards mutated MCL1. Residue-wise interaction network analysis showed the individual residues in Bax binding responsible for the change in stability and interaction due to the protein mutation. In conclusion, the overall findings from the study reveal that the presence of V220F mutation on MCL1 is responsible for the structural confirmational change leading to disruption of its biological functions which might be responsible for tumorigenesis. The mutation could possibly be used as future diagnostic markers in treating cancers.

Host cell proteins (HCPs) are a significant class of process-related impurities commonly associated with the manufacturing of biopharmaceuticals. However, due to the increased use of crude enzymes as biocatalysts for modern organic synthesis, HCPs can also be introduced as a new class of impurities in chemical drugs. In both cases, residual HCPs need to be adequately removed to ensure product purity, quality, and patient safety. Although a lot of attentions have been focused on defining a universally acceptable limit for such impurities, the risks associated with residual HCPs on product quality, safety, and efficacy often need to be determined on a case-by-case basis taken into consideration of residual HCP profile in the product, the dose, dosage form, and administration route etc. Here we describe the unique challenges for residual HCP control presented by the biocatalytic synthesis of a Merck investigational stimulator of interferon genes protein (STING) agonist, MK-1454, which is a cyclic dinucleotide synthesized using E. coli cell lysate overexpressing cyclic GMP-AMP synthase (cGAS) as a biocatalyst. In this study, a holistic characterization of residual protein impurities using a variety of analytical tools, together with in silico immunogenicity prediction of identified proteins, facilitated risk assessment and guided process development to achieve adequate removal of residual protein impurities in MK-1454 API.

Retinal arterial macroaneurysm shows rapid vision loss when rupture occurs; therefore, preventive photocoagulation should be considered, if necessary.

A 22-year male presented with complaints of dyspnea. Multimodality imaging revealed a polypoidal right atrial mass with sub-massive pulmonary embolism. The patient underwent urgent surgery. The pathological examination confirmed it as cardiac myxoma. Cardiac myxoma, a most common primary cardiac tumor, is commonly found in the left atrium. The right atrium is an uncommon site and the usual mode of presentation is the tumor or thrombus embolisation to the pulmonary circulation.

Patients with relapsed follicular lymphoma who do not respond to CAR-T have a poor outcome. We present a case of refractory follicular lymphoma who relapsed after two CAR-T infusions and achieved a complete remission after treatment with obinutuzumab and lenalidomide. This represents a promising treatment option in the post-CAR-T setting.

A 73-year-old man underwent a stage Ⅱ sigmoid colon cancer resection. Careful monitoring was undertaken, and no evidence of metastasis or anastomotic recurrence was noted until 51 months later. Anal examination revealed a pink, firm perianal tumor histologically identified as a metastasis of the colon adenocarcinoma.

Error-in-variables model (EVM) methods require information about input and output measurement variances when estimating model parameters. In EVM, using replicate experiments for estimating output measurement variances is complicated, because true values of inputs may be different when multiple attempts are made to repeat an experiment. To address this issue, we categorize attempted replicate experiments as: i) true replicates (TRs) when uncertain inputs are the same in replicated runs and ii) pseudo-replicates (PRs) when measured inputs are the same, but unknown true values of inputs are different. We propose methodologies to obtain output measurement variance estimates and associated parameter estimates for both situations. We also propose bootstrap methods for obtaining joint-confidence information for the resulting parameter estimates. A copolymerization case study is used to illustrate the proposed techniques. We show that different assumptions noticeably affect the uncertainties in the resulting reactivity-ratio estimates.

Primary invasive breast carcinoma with neuroendocrine differentiation is an uncommon presentation. We hereby report a case diagnosed as invasive ductal carcinoma with neuroendocrine differentiation in a 52-year-old female who presented with a painless right breast lump.

A document by Hanshe Lim. Click on the document to view its contents.

A maturing body of evidence suggests that anthropogenic impacts on river-wetland corridors may be greater and more widespread than previously recognized. We applied the Geomorphic Grade Line (GGL) method to define pre-Anthropocene valley surfaces within segments of the 42-kilometer Entiat River Valley (ERV) of the North Cascade Mountains, USA. We developed GGL-relative elevation models (GGL-REMs) by subtracting, from high-resolution digital elevation data, a detrending surface based on relic fluvial features of the valley floor. We validated the GGL-REMs using surficial geologic maps, C 14-dated soil profiles, and the identifiable remnants of historic dams. We interpreted these data in the context of settlement land use practices including channelization, large wood removal, and beaver ( Castor canadensis) trapping. Our analysis indicates that since the early 20 th century, the river has incised more than two meters in many areas. This triggered a rapid state and process change, wherein unconfined and partially-confined valleys transitioned from net deposition to erosion and transport environments. The distribution of river types shifted from ecologically rich river-wetland corridors towards simpler, single-threaded channels common in confined valleys. The effects of this state change on salmon productivity were profound. Results from the Entiat and other locales indicate that GGL-REMs can be used to help define the fluvial process-form domains, including the vertical dimension needed to guide valley floor restoration. These tools can be used to envisage pre-degradation riverscapes, especially when used in concert with other datasets. Once the pre-Anthropogenic conditions of rivers like Entiatqua have been recognized, the case for restoring lost river-wetland corridors to unlock their ecological potential becomes compelling.

Anthropogenic activities are leading to changes in the environment at global scales, and understanding these changes requires rapid, high-throughput methods of assessment. Pollen DNA metabarcoding and related methods provide advantages in throughput and efficiency over traditional methods, such as microscopic identification of pollen and visual observation of plant-pollinator interactions. Pollen DNA metabarcoding is currently being applied to assessments of plant-pollinator interactions and their responses to land-use change such as increased agricultural intensity and urbanisation, surveillance of ecosystem change, and monitoring of spatiotemporal distribution of allergenic pollen. In combination with historical specimens, pollen DNA metabarcoding can compare contemporary and past ecosystems. Current technical challenges with pollen DNA metabarcoding include the need to understand the relationship between sequence read and species abundance, develop methods for determining confidence limits for detection and taxonomic classification, increase method standardisation, and improve of gaps in reference databases. Future research expanding the method to intraspecific identification, analysis of DNA in ancient pollen samples, and increased use of museum and herbarium specimens could open further avenues for research. Ongoing developments in sequencing technologies can accelerate progress towards these goals. Global ecological change is happening rapidly, and we anticipate that high-throughput methods such as pollen DNA metabarcoding are critical for assessing these changes and providing timely management recommendations to preserve biodiversity and the evolutionary and ecological processes that support it.