Airway tissue engineering for congenital laryngotracheal disease

Regenerative medicine offers hope of a sustainable solution for severe airway disease by the creation of functional, immunocompatible organ replacements. When considering fetuses and newborns, there is a specific spectrum of airway pathologies that could benefit from cell therapy and tissue engineering applications. While hypoplastic lungs associated with congenital diaphragmatic hernia (CDH) could benefit from cellular based treatments aimed at ameliorating lung function, patients with upper airway obstruction could take advantage from a de novo tissue engineering approach. Moreover, the international acceptance of the EXIT procedure as a means of securing the precarious neonatal airway, together with the advent of fetal surgery as a method of heading off postnatal co-morbidities, offers the revolutionary possibility of extending the clinical indication for tissue-engineered airway transplantation to infants affected by diverse severe congenital laryngotracheal malformations. This article outlines the necessary basic components for regenerative medicine solutions in this potential clinical niche

COVID-19 and vertical transmission: assessing the expression of ACE2 / TMPRSS2 in the human fetus and placenta to assess the risk of SARS-CoV-2 infection

Background: While pregnant women have been identified as a potentially at-risk group concerning COVID-19 infection, little is known regarding the susceptibility of the fetus to infection. Co-expression of ACE2 and TMPRSS2 has been identified as a pre-requisite for infection, and expression across different tissues is known to vary between children and adults. However, the expression of these proteins in the fetus is unknown. Methods: We performed a retrospective analysis of single cell data repositories. This data was then validated at both gene and protein level by performing qRT-PCR and two-colour immunohistochemistry on a library of second-trimester human fetal tissues. Findings: TMPRSS2 is present at both gene and protein level in the predominantly epithelial fetal tissues analysed. ACE2 is present at significant levels, only in the fetal intestine and kidney and is not expressed in the fetal lung. The placenta is also negative for the two proteins both during development and at term. Interpretation: This dataset indicates that the lungs are unlikely to be a viable route of SARS-CoV2 fetal infection. The fetal kidney, despite presenting both the proteins required for the infection, is anatomically protected from the exposure to the virus. However, the GI tract is likely to be susceptible to infection due to its high co-expression of both proteins, as well as its exposure to potentially infected amniotic fluid

Preservation of micro-architecture and angiogenic potential in a pulmonary acellular matrix obtained using intermittent intra-tracheal flow of detergent enzymatic treatment

Tissue engineering of autologous lung tissue aims to become a therapeutic alternative to transplantation. Efforts published so far in creating scaffolds have used harsh decellularization techniques that damage the extracellular matrix (ECM), deplete its components and take up to 5 weeks to perform. The aim of this study was to create a lung natural acellular scaffold using a method that will reduce the time of production and better preserve scaffold architecture and ECM components. Decellularization of rat lungs via the intratracheal route removed most of the nuclear material when compared to the other entry points. An intermittent inflation approach that mimics lung respiration yielded an acellular scaffold in a shorter time with an improved preservation of pulmonary micro-architecture. Electron microscopy demonstrated the maintenance of an intact alveolar network, with no evidence of collapse or tearing. Pulsatile dye injection via the vasculature indicated an intact capillary network in the scaffold. Morphometry analysis demonstrated a significant increase in alveolar fractional volume, with alveolar size analysis confirming that alveolar dimensions were maintained. Biomechanical testing of the scaffolds indicated an increase in resistance and elastance when compared to fresh lungs. Staining and quantification for ECM components showed a presence of collagen, elastin, GAG and laminin. The intratracheal intermittent decellularization methodology could be translated to sheep lungs, demonstrating a preservation of ECM components, alveolar and vascular architecture. Decellularization treatment and methodology preserves lung architecture and ECM whilst reducing the production time to 3 h. Cell seeding and in vivo experiments are necessary to proceed towards clinical translation

Virosome-Formulated Plasmodium falciparum AMA-1 & CSP Derived Peptides as Malaria Vaccine: Randomized Phase 1b Trial in Semi-Immune Adults & Children

BACKGROUND\ud \ud This trial was conducted to evaluate the safety and immunogenicity of two virosome formulated malaria peptidomimetics derived from Plasmodium falciparum AMA-1 and CSP in malaria semi-immune adults and children.\ud \ud METHODS\ud \ud The design was a prospective randomized, double-blind, controlled, age-deescalating study with two immunizations. 10 adults and 40 children (aged 5-9 years) living in a malaria endemic area were immunized with PEV3B or virosomal influenza vaccine Inflexal®V on day 0 and 90.\ud \ud RESULTS\ud \ud No serious or severe adverse events (AEs) related to the vaccines were observed. The only local solicited AE reported was pain at injection site, which affected more children in the Inflexal®V group compared to the PEV3B group (p = 0.014). In the PEV3B group, IgG ELISA endpoint titers specific for the AMA-1 and CSP peptide antigens were significantly higher for most time points compared to the Inflexal®V control group. Across all time points after first immunization the average ratio of endpoint titers to baseline values in PEV3B subjects ranged from 4 to 15 in adults and from 4 to 66 in children. As an exploratory outcome, we found that the incidence rate of clinical malaria episodes in children vaccinees was half the rate of the control children between study days 30 and 365 (0.0035 episodes per day at risk for PEV3B vs. 0.0069 for Inflexal®V; RR  = 0.50 [95%-CI: 0.29-0.88], p = 0.02).\ud \ud CONCLUSION\ud \ud These findings provide a strong basis for the further development of multivalent virosomal malaria peptide vaccines.\ud \ud TRIAL REGISTRATION\ud \ud ClinicalTrials.gov NCT00513669

Simons Observatory: Constraining inflationary gravitational waves with multitracer B-mode delensing

We introduce and validate a delensing framework for the Simons Observatory (SO), which will be used to improve constraints on inflationary gravitational waves by reducing the lensing noise in measurements of the&nbsp;B&nbsp;modes in CMB polarization. SO will initially observe CMB by using three small aperture telescopes and one large-aperture telescope. While polarization maps from small-aperture telescopes will be used to constrain inflationary gravitational waves, the internal CMB lensing maps used to delens will be reconstructed from data from the large-aperture telescope. Since lensing maps obtained from the SO data will be noise dominated on subdegree scales, the SO lensing framework constructs a template for lensing-induced&nbsp;B&nbsp;modes by combining internal CMB lensing maps with maps of the cosmic infrared background from Planck as well as galaxy density maps from the LSST survey. We construct a likelihood for constraining the tensor-to-scalar ratio&nbsp;r&nbsp;that contains auto and cross spectra between observed&nbsp;B&nbsp;modes and the lensing&nbsp;B-mode template. We test our delensing analysis pipeline on map-based simulations containing survey nonidealities, but that, for this initial exploration, does not include contamination from Galactic and extragalactic foregrounds. We find that the SO survey masking and inhomogeneous and atmospheric noise have very little impact on the delensing performance, and the&nbsp;r&nbsp;constraint becomes&nbsp;&sigma;(r)&asymp;0.0015&nbsp;which is close to that obtained from the idealized forecasts in the absence of the Galactic foreground and is nearly a factor of 2 tighter than without delensing. We also find that uncertainties in the external large-scale structure tracers used in our multitracer delensing pipeline lead to bias much smaller than the&nbsp;1&sigma;&nbsp;statistical uncertainties.</p

Simons Observatory: Constraining inflationary gravitational waves with multitracer B -mode delensing

We introduce and validate a delensing framework for the Simons Observatory (SO), which will be used to improve constraints on inflationary gravitational waves by reducing the lensing noise in measurements of the B modes in CMB polarization. SO will initially observe CMB by using three small aperture telescopes and one large-aperture telescope. While polarization maps from small-aperture telescopes will be used to constrain inflationary gravitational waves, the internal CMB lensing maps used to delens will be reconstructed from data from the large-aperture telescope. Since lensing maps obtained from the SO data will be noise dominated on subdegree scales, the SO lensing framework constructs a template for lensing-induced B modes by combining internal CMB lensing maps with maps of the cosmic infrared background from Planck as well as galaxy density maps from the LSST survey. We construct a likelihood for constraining the tensor-to-scalar ratio r that contains auto and cross spectra between observed B modes and the lensing B-mode template. We test our delensing analysis pipeline on map-based simulations containing survey nonidealities, but that, for this initial exploration, does not include contamination from Galactic and extragalactic foregrounds. We find that the SO survey masking and inhomogeneous and atmospheric noise have very little impact on the delensing performance, and the r constraint becomes σ(r)≈0.0015 which is close to that obtained from the idealized forecasts in the absence of the Galactic foreground and is nearly a factor of 2 tighter than without delensing. We also find that uncertainties in the external large-scale structure tracers used in our multitracer delensing pipeline lead to bias much smaller than the 1σ statistical uncertainties

A public antibody lineage that potently inhibits malaria infection through dual binding to the circumsporozoite protein.

Immunization with attenuated Plasmodium falciparum sporozoites (PfSPZs) has been shown to be protective against malaria, but the features of the antibody response induced by this treatment remain unclear. To investigate this response in detail, we isolated IgM and IgG monoclonal antibodies from Tanzanian volunteers who were immunized with repeated injection of Sanaria PfSPZ Vaccine and who were found to be protected from controlled human malaria infection with infectious homologous PfSPZs. All isolated IgG monoclonal antibodies bound to P. falciparum circumsporozoite protein (PfCSP) and recognized distinct epitopes in its N terminus, NANP-repeat region, and C terminus. Strikingly, the most effective antibodies, as determined in a humanized mouse model, bound not only to the repeat region, but also to a minimal peptide at the PfCSP N-terminal junction that is not in the RTS,S vaccine. These dual-specific antibodies were isolated from different donors and were encoded by VH3-30 or VH3-33 alleles that encode tryptophan or arginine at position 52. Using structural and mutational data, we describe the elements required for germline recognition and affinity maturation. Our study provides potent neutralizing antibodies and relevant information for lineage-targeted vaccine design and immunization strategies

The Simons Observatory: Gain, bandpass and polarization-angle calibration requirements for B-mode searches

We quantify the calibration requirements for systematic uncertainties for next-generation ground-based observatories targeting the large-angle B-mode polarization of the Cosmic Microwave Background, with a focus on the Simons Observatory (SO). We explore uncertainties on gain calibration, bandpass center frequencies, and polarization angles, including the frequency variation of the latter across the bandpass. We find that gain calibration and bandpass center frequencies must be known to percent levels or less to avoid biases on the tensor-to-scalar ratio r on the order of Δ r∼10-3, in line with previous findings. Polarization angles must be calibrated to the level of a few tenths of a degree, while their frequency variation between the edges of the band must be known to O(10) degrees. Given the tightness of these calibration requirements, we explore the level to which residual uncertainties on these systematics would affect the final constraints on r if included in the data model and marginalized over. We find that the additional parameter freedom does not degrade the final constraints on r significantly, broadening the error bar by O(10%) at most. We validate these results by reanalyzing the latest publicly available data from the collaboration within an extended parameter space covering both cosmological, foreground and systematic parameters. Finally, our results are discussed in light of the instrument design and calibration studies carried out within SO