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Mathematics is concerned with the method used in the teaching and learning process in addition to issues encountered in the cognitive domain. The Philippines’ education system is still dominated by traditional mathematics teaching, which frequently overlooks the goal of mathematics education—to prepare students to deal successfully with real-life situations. This affects the declining performance of the students in their overall mathematical ability, especially in problem-solving. Hence, this study utilized a pre-experimental design to measure the effectiveness of the Realistic Mathematics Education (RME) approach in the problem-solving skills of the students in terms of understanding the problem, devising a plan, carrying out the plan, and looking back. Furthermore, the cluster sampling technique was used in choosing thirty-five grade 9 students and evaluated their problem-solving ability using a pre-test and post-test assessments. Based on the result, there is a highly significant difference in the mean pre-test and post-test performance of the respondent before and after using the RME approach in all the four phases of problem-solving (p-value=0.000). This implies that the RME is an effective teaching approach that successfully improved the mathematical proficiency of the students, especially in all aspects of problem-solving skills. The findings verify that educators can use the RME approach to expose their students to more collaborative teaching-learning processes that incorporate real-world scenarios. Future researchers may also conduct a similar study in face-to-face learning to comprehensively use the RME approach.

Background: Face masks reduce disease transmission by protecting the wearer from inhaled pathogens and reducing the emission of infectious aerosols. Although methods quantifying efficiency for wearer protection are established, current methods for assessing face mask containment efficiency rely on measurement of a low concentration of aerosols emitted from an infected or noninfected individual. Methods: A small port enabled the introduction of 0.05 µm sodium chloride particles at a constant rate behind the mask worn by a study participant. A condensation particle counter monitored ambient particle numbers 60 cm in front of the participant over 3-minute periods of rest, speaking, and coughing. The containment efficiency (%) for each mask and procedure was calculated as follows: 100 × (1 − average ambient concentration with face covering worn/average ambient concentration with a sham face covering in place). The protection efficiency (%) was also measured using previously published methods. The probability of transmission (%) from infected to uninfected (a function of both the containment efficiency and the protection efficiency) was calculated as follows: {1 − (containment efficiency/100)}×{1 − (protection efficiency/100)}×100. Results: The average containment efficiencies for each mask over all procedures and repeated measures were 94.6%, 60.9%, 38.8%, and 43.2%, respectively, for the N95 mask, the KN95 mask, the procedure face mask, and the gaiter. The corresponding protection efficiencies for each mask were 99.0%, 63.7%, 45.3%, and 24.2%, respectively. For example, the transmission probability for 1 infected and 1 uninfected individual in close proximity was ∼14.2% for KN95 masks, compared to 36%–39% when only 1 individual wore a KN95 mask. Conclusion: Overall, we detected a good correlation between the protection and containment that a face covering afforded to a wearer.

In urban areas, numerous barriers exist for children and families to access needed behavioral health care. Compounding the general deficit of behavioral health workers in the United States is lack of access to culturally responsive care. Additional challenges include inherent racism and oppression in our health and human service systems and siloed approaches to behavioral health care training and practice. Integrated care is an emerging field nationally, yet training and education in integrated care is limited. This article provides an overview of the Child/Adolescent Interprofessional Practice and Education (CAIPE) program, a novel training approach in its first year of implementation for behavioral health students that prioritizes trauma-informed and social-justice oriented practice, integrated care, and interprofessional teamwork.This study employs a mixed-methods approach to program evaluation. Data sources include program application materials (demographics and data related to clinical interests, student goals, and preparedness for interprofessional work). Data is also drawn from measures of student outcomes (self-efficacy navigating complex care patients, empathy, and complex care knowledge).Baseline data are reported for demographics, students' goals and interprofessional interests, and student outcomes. Information is also presented relevant to trainings conducted in the grant's first year and planned for subsequent years. Preliminary data and implementation suggest that the program has drawn diverse trainees who are committed to interprofessional, integrated care for underserved youth.Recruitment and program development challenges are discussed, along with grant goals' fit with students' interests and plans for future evaluation. (PsycInfo Database Record (c) 2022 APA, all rights reserved).

Total and regional deposition of inhaled electronic cigarette (E-cig) particles in the respiratory tract (RT) depends on both physical properties of the inhaled particles and biological factors of users, for example, breathing pattern or puff profile, airway anatomy, and regional ventilation. Accurate particle sizing of E-cig aerosols is essential for predicting particle deposition in the RT. Studies using a variety of sizing methods have shown mass median aerodynamic diameters ranging from 0.2 to 1.2 um and secondary count diameters in the ultrafine range (0.1 μm). Incorporating these particle sizes into a multiple-path particle dosimetry (MPPD) model shows 10% to 45% total lung deposition by mass and 30% to 80% for ultrafine particles depending on the breathing patterns. These predictions are consistent with experimental measures of deposition fraction of submicron and ultrafine particles. While box-mod-type E-cig devices allow for full "direct-lung" inhalations of aerosol, the more recent pod-based, and disposable E-cigs (e.g., JUUL, Puff Bar, Stig) deliver the aerosol as a "mouth-to-lung" puff, or bolus, that is inhaled early in the breath followed to various degrees by further inhalation of ambient air. Measurement of realistic ventilation patterns associated with these various devices may further improve deposition predictions. Finally, while in vivo measures of RT deposition present a challenge, a recent methodology to radiolabel E-cig particles may allow for such measurements by gamma scintigraphy. Supported by NIH/NHLBI R01HL139369. © 2022 American Physiological Society. Compr Physiol 12: 1-10, year.

Three of the most prevalent developmental disorders (DDs) are autism spectrum disorder (ASD), attention-deficit/hyperactivity disorder (ADHD) and fetal alcohol spectrum disorder (FASD). As part of a study screening for DDs in Greater Manchester, UK, a unique opportunity was taken to explore parents' experiences of receiving reports about their child's previously unrecognised DD. Six parents out of a possible 16 took part in interviews, which were analysed thematically. Findings drawn from parental responses revealed a number of barriers to accessing support for their child's additional needs, including perceived resistance from schools, particularly for quiet, well-behaved girls, and difficulty in accessing assessment or referrals. There needs to be greater awareness of additional needs in children without externalising behaviours, the presence of gender-specific differences in the presentation of DDs, and FASD as a commonly occurring DD. Ultimately, better support for children with DDs would reduce the burden of unmet needs for the children and their families, and for wider services.

Vegetation patterns during salt marsh restoration reflect underlying processes related to colonization, reproduction, and interactions of halotolerant plants. Examining both pattern and process during recovery is valuable for understanding and managing salt marsh restoration projects. We present a decade of vegetation dynamics during salt marsh restoration (2011–2020) at a study site in the Bay of Fundy with megatidal amplitudes, strong currents, cold winter temperatures, and ice. We mainly investigated reproduction (asexual and sexual) and associated spread rates of Spartina grasses, and their health-related states (stem density, canopy height, and percent flowering) which help inform the probability of processes occurring. We also estimated modes of colonization and began quantifying the effects of interspecific interactions and environmental conditions on plant state. Spartina pectinata was the only pastureland plant to survive dike-breaching and saltwater intrusion in 2010; however, it was stunted compared to reference plants. Spartina pectinata patches remained consistent initially, before decreasing in size, and disappearing by the fifth year (2015). This early dynamic may provide initial protection to a developing salt marsh before Spartina alterniflora becomes established. Spartina alterniflora first colonized the sites in year 2 (2012), likely via deposition of rhizomal material, and then spread asexually before seedlings (sexual reproduction) appeared in year 4 (2014). Vegetation cover subsequently increased greatly until near-complete in year 9 (2019). The early successional dynamics of S. pectinata and S. alterniflora occurred spatially independently of each other, and likely contributed to sediment retention, creating an improved environment for S. patens, the dominant high marsh species in our region. Spartina patens have been slowly spreading into restoration sites from high elevation areas since year 6 (2016). We expect that competition between S. alterniflora and S. patens will result in the typical distinct zonation between high and low marsh zones. A next study will use the quantified processes for spatial-explicit modeling to simulate patterns of vegetation recovery, and to evaluate different salt marsh restoration strategies for the Bay of Fundy and elsewhere. Thus, proper identification and quantification of pattern-building processes in salt marsh vegetation recovery, the focus of our present study, was an essential step.

High-quality peer-reviewer training open to researchers across the globe has the potential to improve the published literature, however, this type of training is not widely available. In this paper, we describe an online peer-reviewer training programme, highlight its effectiveness in building peer review and writing skills, and discuss challenges and lessons learnt. This training programme, open to researchers across the globe, acquaints participants with challenges to and inequities in publishing and educates them about writing effective peer reviews. A focal point is how to provide specific and respectful feedback to help authors get accepted for peer review at an academic journal. Forty-nine participants from or residing in six continents completed the training. All programme evaluation respondents agreed that the orientation helped them gain a better understanding of their role as a peer reviewer at Pre-Publication Support Service. Most agreed that the training was helpful in improving their peer-review skills, and that the training was helpful in improving their writing skills. Participants wanted more networking and collaboration opportunities with other peer reviewers, inclusion of a qualitatively researched example paper and improved communication about the required time commitment. Our online programme with multiple time options was geographically inclusive but internet connectivity was challenging for some participants. Peer-reviewer training programmes can help researchers build their peer review and writing skills and enhance participants’ understanding of disparities in publishing. Integrating a geographically diverse group of researchers has the potential to enrich the discussions and learning in such a programme.

Facial feminization surgery (FFS) is an integral aspect of gender-affirming surgery (GAS) for individuals seeking to align secondary sex characteristics and gender identity. Despite the importance of FFS in treating gender dysphoria, current trends and prevalence remain unknown. We sought to examine trends in GAS and FFS and investigate the payer status of facial feminization procedures in the United States.Data was extracted from the National Inpatient Sample from 2008 to 2017 by using International Classification of Diseases Ninth or Tenth diagnosis codes for gender identity disorder and procedure codes for FFS.From 2008 to 2017, 3015 patients underwent GAS. The yearly number of cases increased as did the average cost of GAS, which rose from $13,657 in 2008 to $50,789 in 2017. From 2015 to 2017, when FFS data was available, 110 of 1215 (9.1%) GAS patients had FFS. Most were non-Hispanic White (66.7%) or Black (23.8%). Fifty percent of FFS cases occurred in the West, followed by the Northeast (31.8%), South (13.6%), and Midwest (4.8%) (From 2008 to 2017, GAS cases increased nationwide while the average cost of surgery rose steeply. FFS cases were primarily in the Western and Northeast United States. Despite high cost, roughly 18% of transgender women in our sample received FFS, highlighting the importance of FFS in gender transition.

Predation is a fundamental ecological process that shapes communities and drives evolutionary dynamics. As the world rapidly urbanizes, it is critical to understand how human perturbations alter predation and meat consumption across taxa. We conducted a meta-analysis to quantify the effects of urban environments on three components of trophic ecology in predators: dietary species richness, dietary evenness and stable isotopic ratios (IRs) ( δ 13 C and δ 15 N IR). We evaluated whether the intensity of anthropogenic pressure, using the human footprint index (HFI), explained variation in effect sizes of dietary attributes using a meta-regression. We calculated Hedges’ g effect sizes from 44 studies including 11 986 samples across 40 predatory species in 39 cities globally. The direction and magnitude of effect sizes varied among predator taxa with reptilian diets exhibiting the most sensitivity to urbanization. Effect sizes revealed that predators in cities had comparable diet richness, evenness and nitrogen ratios, though carbon IRs were more enriched in cities. We found that neither the 1993 nor 2009 HFI editions explained effect size variation. Our study provides, to our knowledge, the first assessment of how urbanization has perturbed predator–prey interactions for multiple taxa at a global scale. We conclude that the functional role of predators is conserved in cities and urbanization does not inherently relax predation, despite diets broadening to include anthropogenic food sources such as sugar, wheat and corn.

Background Hypertonic saline (HS) is commonly prescribed for children with cystic fibrosis (CF) despite the absence of strong data indicating clinical efficacy in a population with mild lung disease. We hypothesized that HS treatment would result in a sustained improvement in mucociliary clearance (MCC) in children with CF who had minimal lung disease, thus providing evidence for a biologically relevant effect that also may be associated with clinical improvements. Methods We performed a randomized, placebo controlled, double blind study of 6% versus 0.12% sodium chloride, delivered three-times daily with an eFlow nebulizer for 4 weeks. MCC was measured using gamma scintigraphy at baseline, 2-hours after the first study treatment, and ~12-hours after the final dose (at day 28). Spirometry, respiratory symptoms (CFQ-R), and safety were also assessed. Results Study treatments were generally well tolerated and safe. HS (6% sodium chloride) resulted in a significant, sustained improvement from baseline in whole lung clearance after 4 weeks of therapy (p = 0.014), despite absence of a prolonged single-dose effect after the initial dose. This sustained change (12 hrs after prior dose) was significantly greater when compared to placebo (0.12% sodium chloride) treatment (p = 0.016). Improvements in spirometry with HS did not reach statistical significance but correlated with MCC changes. Conclusions The observed sustained improvement in MCC with HS suggests that this treatment may yield health benefits, even in relatively mildly affected children with CF. Highlighting this physiologic finding is important due to the lack of meaningful, validated endpoints in this population.

Background Mucus dehydration and impaired mucus clearance are common features of cystic fibrosis (CF) and chronic obstructive pulmonary disease (COPD). In CF, inhaled hypertonic saline (HS) improves lung function and produces sustained increases in mucociliary clearance (MCC). We hypothesised that administration of HS (7% NaCl) twice daily for 2 weeks would improve clinical outcomes and produce sustained increases in MCC in COPD subjects with a chronic bronchitis (CB) phenotype. Methods Twenty-two CB subjects completed a double-blinded, crossover study comparing inhaled HS to a hypotonic control solution (0.12% saline) administered via nebuliser twice daily for 2 weeks. Treatment order was randomised. During each treatment period, symptoms and spirometry were measured. MCC was measured at baseline, shortly after initial study agent administration, and approximately 12 h after the final dose. Results HS was safe and well tolerated but overall produced no significant improvements in spirometry or patient-reported outcomes. CB subjects had slower baseline MCC than healthy subjects. The MCC rates over 60 min (Ave60Clr) in CB subjects following 2 weeks of HS were not different from 0.12% saline but were slower than baseline (Ave60Clr was 9.1±6.3% at baseline versus 5.3±6.9% after HS; p, 2-week HS treatment in CB does not lead to overall improvement in baseline mucociliary clearance. However, a subgroup of patients with residual, noncough-induced central airway clearance does show improvement in both spirometry and symptoms. https://bit.ly/36ji6Vn

Each step in the literature search phase of the meta-analysis resulted in either inclusion or exclusion of each study, the results of which are summarized here in accordance with PRISMA guidelines.

International audience; Primary ciliary dyskinesia (PCD) is a genetic disorder in which impaired ciliary function leads to chronic airway disease. Exome sequencing of a PCD subject identified an apparent homozygous frameshift variant, c.887_890delTAAG (p.Val296Glyfs∗13), in exon 5; this frameshift introduces a stop codon in amino acid 308 of the growth arrest-specific protein 2-like 2 (GAS2L2). Further genetic screening of unrelated PCD subjects identified a second proband with a compound heterozygous variant carrying the identical frameshift variant and a large deletion (c.867_∗343+1207del; p.?) starting in exon 5. Both individuals had clinical features of PCD but normal ciliary axoneme structure. In this research, using human nasal cells, mouse models, and X.laevis embryos, we show that GAS2L2 is abundant at the apical surface of ciliated cells, where it localizes with basal bodies, basal feet, rootlets, and actin filaments. Cultured GAS2L2-deficient nasal epithelial cells from one of the affected individuals showed defects in ciliary orientation and had an asynchronous and hyperkinetic (GAS2L2-deficient = 19.8 Hz versus control = 15.8 Hz) ciliary-beat pattern. These results were recapitulated in Gas2l2-/- mouse tracheal epithelial cell (mTEC) cultures and in X. laevis embryos treated with Gas2l2 morpholinos. In mice, the absence of Gas2l2 caused neonatal death, and the conditional deletion of Gas2l2 impaired mucociliary clearance (MCC) and led to mucus accumulation. These results show that a pathogenic variant in GAS2L2 causes a genetic defect in ciliary orientation and impairs MCC and results in PCD

This cohort study examines changes in gender-affirming surgery by US region and insurance use during periods when antidiscrimination policies took effect in the US between 2008 and 2017.

Although global vaccination efforts against SARS-CoV-2 are underway, the public is urged to continue using face masks as a primary intervention to control transmission. Recently, US public health officials have also encouraged doubling masks as a strategy to counter elevated transmission associated with infectious SARS-CoV-2 variants. US Centers for Disease Control and Prevention investigators reported that doubling masks increased effectiveness, but their assessment was limited in type and combinations of masks tested, as well as by the use of head forms rather than humans. To address these limitations, this study compared the fitted filtration efficiency (FFE) of commonly available masks worn singly, doubled, or in combinations.

Shortages of efficient filtering facepiece respirators leave the public vulnerable to transmission of infectious diseases in small particle aerosols. This study demonstrates that a high-filtration-efficiency facepiece capable of filtering out >95% of 0.05μm particles while being worn can be simply produced with available materials.

Background Globally, a large percentage of men keep a beard at least occasionally. Workplace regulations prohibit beards with N95 respirators, but there is little information on the effect of beards with face masks worn by the public for protection against SARS-CoV-2. Methods and findings We examined the fitted filtration efficiency (FFE) of five commonly worn protective face masks as a function of beard length following the US Occupational Safety and Health Administration Quantitative Fit Test: N95 (respirator), KF94 and KN95, surgical/procedure, and cloth masks. A comparison using N95 respirators was carried out in shaven and bearded men. A detailed examination was conducted for beard lengths between 0 and 10 mm (0.5 mm increments). The effect of an exercise band covering the beard on FFE was also tested. Although N95 respirators showed considerable variability among bearded men, they had the highest FFE for beard lengths up to 10 mm. KF94 and KN95 masks lost up to 40% of their FFE. Procedure and cotton masks had poor performance even on bare skin (10–30% FFE) that did not change appreciably with beard length. Marked performance improvements were observed with an exercise band worn over the beard. Conclusions Though variable, N95 respirators offer the best respiratory protection for bearded men. While KF94 and KN95 FFE is compromised considerably by increasing beard length, they proved better options than procedure and cotton face masks. A simple exercise band improves FFE for face masks commonly used by bearded men during the COVID-19 pandemic.

Predation is a fundamental ecological process that shapes communities and drives long-term evolutionary dynamics. As the world rapidly urbanizes, it is critical to understand how the built environment and other human perturbations alter predation across taxa. We conducted a meta-analysis to quantify the effects of urban environments on three components of trophic ecology in predators: dietary species richness (DSR), dietary evenness (DEV), and stable isotopic ratios (δ13C and δ15N IR). We then evaluated whether intensity of anthropogenic pressure, using the human footprint index (HFI), explained variation in the effect sizes of dietary attributes using a meta-regression. We calculated Hedges’ g effect sizes from 44 studies including 11,986 samples across 40 predatory species in 39 cities globally. The direction and magnitude of effect sizes varied between predator taxonomic groups with reptile diets exhibiting the most sensitivity to urbanization. Effect sizes revealed that predators in cities had comparable DSR, DEV, and nitrogen ratios, though carbon consumption was significantly higher. We found that HFI did not explain variation in effect sizes, a result consistent between the 1993 and 2009 editions of this metric. Our study provides the first assessment of how urbanization has perturbed predator-prey interactions for multiple taxa at a global scale, revealing that the functional role of predators is conserved in cities and urbanization does not inherently relax predation.

Breast implant surgery remains one of the most common surgical procedures performed in the United States. Implant exchange can be complicated by unavailability of medical records or implant identification cards. Using chest imaging of 154 breast implants, an algorithm for estimating breast implant volume was generated. Based on four simple measurements and patient body mass index, a free, online calculator was created with a mean error of volume estimate of less than 1 cm

Wildlife adaptively respond to human presence by adjusting their temporal niche, possibly modifying encounter rates among species and trophic dynamics that structure communities. Here we show that these human-induced modifications to behaviours are prolific among species and alter apex predators’ access to prey resources. We assessed human-induced changes to wildlife diel activity and consequential changes in predator-prey overlap using 11,954 detections of three apex predators and 13 ungulates across 21,430 trap-nights in West Africa. Over two-thirds of species altered their diel use in response to human presence, and ungulate nocturnal activity increased by 6.8%. Rather than traditional pairwise predator-prey comparisons, we considered spatiotemporally explicit predator access to a suite of prey resources to evaluate community-level trophic responses to human presence. Although leopard prey access was not affected, lion and hyena access to 3 prey species significantly increased when prey increased their nocturnal temporal niche to avoid humans. Ultimately, humans considerably altered the composition of available prey, with implications for prey selection, demonstrating how humans perturb ecological processes via behavioural modifications.

Background: Using planar gamma scintigraphy of inhaled radioaerosols, we have developed new analytical methods for assessing homogeneity of aerosol deposition and time-dependent particle clearance on a pixel-by-pixel basis, and applied them to a therapeutic cystic fibrosis (CF) study. Methods: At baseline and 1 month after beginning treatment with ivacaftor, a cystic fibrosis transmembrane regulator modulator for CF patients with at least one copy of the G551D mutation (n = 13), initial deposition and subsequent mucociliary clearance (MCC) of radiolabeled particles ((99m)Technetium-sulfur colloid, 5 μm mass median aerodynamic diameter) inhaled under controlled breathing conditions were measured. Results: Improved homogeneity of deposition, that is, decreased areas of higher and lower particle deposition in the lungs, was observed following ivacaftor treatment. The mean number ratio (NR) of pixels with higher deposition, relative to lung size, decreased from 0.14 to 0.09 (p = 0.003) and mean NR of colder pixels decreased from 0.23 to 0.19 (p = 0.004). Particle clearance was also improved following treatment, with mean MCC through 60 minutes equal to 12% versus 24%, without and with treatment, respectively (p = 0.010). Pixel-level analysis of MCC showed that (1) the fraction of pixels clearing >30% at 60 minutes was increased from 0.13 to 0.32 (p = 0.007); and (2) the fraction of pixels clearing

Efficient delivery of aerosols to the lungs via the nasal route has been difficult to achieve, but it may offer benefits over the traditional oral route for a range of patient populations. Because slow, continuous delivery of short-acting agents could improve safety, tolerability, compliance, and efficacy when compared with the rapid, intermittent aerosol treatments delivered by mouthpiece or mask, a novel trans-nasal pulmonary aerosol delivery (tPAD) device was developed. The tPAD incorporates an aerosol particle-size selection chamber and a custom nasal cannula that are specifically optimized for aerosol delivery to the lung via the nasal route. The tPAD device produced a steady aerosol output (∼2 mL/h) from an optimized nasal cannula with negligible rainout in the cannula for up to 8 hours. The generated aerosol particles were small enough to minimize nasal deposition [volume median diameter (VMD) = 1.4 μm].In this proof-of-concept study, gamma scintigraphy was used to quantitate deposition efficiency ofThe tPAD device achieved high pulmonary deposition (39% ± 8%), based on emitted dose, and matched that of the oral jet nebulizer (36% ± 9%). Low fractions of aerosol deposition in the head and nose region were observed for tPAD (6% ± 6%) and jet nebulizer deliver (1% ± 1%) as well.A profile of high pulmonary deposition efficiency and low nasal dose may enable the sustained use of the tPAD platform with a variety of therapeutic agents for a range of pulmonary disorders.

Background The ability to restore cystic fibrosis transmembrane regulator (CFTR) function with effective small molecule modulators in patients with cystic fibrosis provides an opportunity to study relationships between CFTR ion channel function, organ level physiology, and clinical outcomes. Methods We performed a multisite, prospective, observational study of ivacaftor, prescribed in patients with the G551D-CFTR mutation. Measurements of lung mucociliary clearance (MCC) were performed before and after treatment initiation (1 and 3 months), in parallel with clinical outcome measures. Results Marked acceleration in whole lung, central lung, and peripheral lung MCC was observed 1 month after beginning ivacaftor and was sustained at 3 months. Improvements in MCC correlated with improvements in forced expiratory volume in the first second (FEV1) but not sweat chloride or symptom scores. Conclusions Restoration of CFTR activity with ivacaftor led to significant improvements in MCC. This physiologic assessment provides a means to characterize future CFTR modulator therapies and may help to predict improvements in lung function. Trial registration ClinicialTrials.gov, NCT01521338. Funding CFF Therapeutics (GOAL11K1).

Importance Procuring respiratory protection for clinicians and other health care workers has become a major challenge of the coronavirus disease 2019 (COVID-19) pandemic and has resulted in nonstandard practices such as the use of expired respirators and various decontamination processes to prolong the useful life of respirators in health care settings. In addition, imported, non–National Institute for Occupational Safety and Health (NIOSH)-approved respirators have been donated or acquired by hospitals as a potential replacement for limited NIOSH-approved N95 respirators. Objective To assess fitted filtration efficiencies (FFEs) for face mask alternatives used during the COVID-19 pandemic. Design, Setting, and Participants For this quality-improvement study conducted between April and June 2020, we used the Occupational Safety and Health Administration’s Quantitative Fit Testing Protocol for Filtering Facepiece Respirators in a laboratory atmosphere supplemented with sodium chloride particles to assess the FFEs of a variety of respirators worn by a male volunteer and female volunteer. Main Outcomes and Measures The FFEs of respirators commonly worn by clinicians and other health care workers and available respirator alternatives during the COVID-19 pandemic. Results Of the 29 different fitted face mask alternatives tested on 1 man and 1 woman, expired N95 respirators with intact elastic straps and respirators subjected to ethylene oxide and hydrogen peroxide sterilization had unchanged FFE (>95%). The performance of N95 respirators in the wrong size had slightly decreased performance (90%-95% FFE). All of the respirators not listed as approved in this evaluation (n = 6) failed to achieve 95% FFE. Neither of the 2 imported respirators authorized for use by the Centers for Disease Control and Prevention that were not NIOSH-approved tested in this study achieved 95% FFE, and the more effective of the 2 functioned at approximately 80% FFE. Surgical and procedural face masks had filtering performance that was lower relative to that of N95 respirators (98.5% overall FFE), with procedural face masks secured with elastic ear loops showing the lowest efficiency (38.1% overall FFE). Conclusions and Relevance This quality-improvement study evaluating 29 face mask alternatives for use by clinicians interacting with patients during the COVID-19 pandemic found that expired N95 respirators and sterilized, used N95 respirators can be used when new N95 respirators are not available. Other alternatives may provide less effective filtration.

Herein we demonstrate the use of metal organic framework layers coated on zeolite 5A adsorbent as effective nano-valves for CH4 storage at low storage pressure. The resultant nanovalved materials are capable of sealing high pressure CH4 inside the adsorbents and storing it at low pressure. Four different metal organic frameworks (MOF) were grown on 5A zeolite beads, and functioned as nanovalves. These MOFs included HKUST-1, Al-MOF, Ga-MOF, and Co-MOF. Ethanol, a cheap, renewable, and safe chemical, was employed as the nanovalve sealing molecule. It was found that the 2-crystallization step MOFs displayed the best performance regardless of the MOF coordinating metal. The methane storage capacity followed the order: Al-MOF > Ga-MOF > Co-MOF > Cu-MOF. The best performing material, 2-crystallization step Al-MOF, was able to store 33.6 (L (STP) CH4/L nanovalve adsorbent) excess CH4, corresponding to 46 % of the CH4 saturation capacity of the zeolite 5A bead adsorbent at

The cellular process responsible for providing energy for most life on Earth, namely, photosynthetic light-harvesting, requires the cooperation of hundreds of proteins across an organelle, involving length and time scales spanning several orders of magnitude over quantum and classical regimes. Simulation and visualization of this fundamental energy conversion process pose many unique methodological and computational challenges. We present, in two accompanying movies, light-harvesting in the photosynthetic apparatus found in purple bacteria, the so-called chromatophore. The movies are the culmination of three decades of modeling efforts, featuring the collaboration of theoretical, experimental, and computational scientists. We describe the techniques that were used to build, simulate, analyze, and visualize the structures shown in the movies, and we highlight cases where scientific needs spurred the development of new parallel algorithms that efficiently harness GPU accelerators and petascale computers.

Understanding the regional partition of deposition of inhaled particles within the lung is important for improving targeted delivery of inhaled aerosolized drugs. One factor affecting regional deposition is gravity. As the lung deforms under its own weight, changes in lung volume, in airway geometries, and in spatial patterns of ventilation distribution between postures have the potential to alter the regional distribution of deposited particles.Using gamma-scintigraphy, we measured regional deposition and clearance of (99m)Tc labeled particles (5 μm) in 6 healthy subjects, with aerosol inhalation occurring both in the supine and seated postures at constant flow (0.5 L/sec) and breathing rate (15 breaths/min). After aerosol deposition, mucociliary clearance data were collected in the seated posture, immediately post-particle administration, 1 h 30 min, 4 h, and 22 h post-inhalation. Relative regional deposition was computed using retention (R) at the different time points, with (1-R(1h30min)), (R(1h30min)- R(4h)), and (R(4h)- R(22h)) corresponding to deposition in the large, intermediate, and small airways, respectively. Alveolar deposition was estimated as the relative retention at 22 h (R(22h)).Relative deposition of coarse particles in the alveolar region decreased from 60±8% seated to 34±16% supine (p=0.04). This change was accompanied by an increase in relative deposition in the intermediate (7±3% seated to 16±17% supine, P=0.09) and small airways (19±6% seated to 34±13% supine, p=0.06) when inhalation occurred in the supine posture. No change was observed in central to peripheral deposition (C/P ratio), the skew of the deposition distribution, or the apex-to-base ratio of deposition between seated and supine postures.Inhalation of coarse particles in the supine posture shifts relative deposition from the alveolar to the bronchial airways, when compared to the seated posture, likely driven by changes in functional residual capacity, and airway size, as well as changes in the regional distribution of ventilation between postures.

Background: Electronic cigarettes (ECIGs) are widely used, but their health effects are not well known. ECIG exposure is difficult to quantify, and a direct measurement of deposition would be beneficial to in vivo and in vitro toxicity studies. The aim of this study was to demonstrate effective radiolabeling of an ECIG. Methods: A technetium-99m-labeled carbon ultrafine (TCU) aerosol was generated and introduced to a fourth-generation ECIG before nucleation and aerosol formation. The aerosolized e-liquid was a commercially available strawberry flavor containing 1.2% nicotine in a 55% propylene glycol and 45% vegetable glycerine base. An ECIG power setting of 100 W was selected. Mass and radioactivity were measured on each stage within a Sierra Cascade Impactor at 14 L/min to verify the labeling technique using the calculated aerodynamic diameters. A strong positive correlation (R(2) > 0.95) between the percent activity and percent mass deposition on each stage provides a reliable validation of colocation. Results: Unlabeled ECIG aerosol from the chosen e-liquid produced a mass median aerodynamic diameter (MMAD) of 0.85 μm. An ECIG labeled with TCU produced an aerosol with an activity median aerodynamic diameter of 0.84 μm and an MMAD of 0.84 μm. The relative mass versus radioactivity on each plate was highly correlated (average R(2) = 0.973, p

Background Observational studies suggest that orally administered guaifenesin (GGE) may thin lower respiratory tract secretions but none have examined its effects on mucociliary and cough clearance (MCC/CC) during a respiratory tract infection (RTI). The current study was a randomized, parallel-group, double-blind, placebo-controlled study in non-smoking adults who suffered from an acute upper RTI. Methods We assessed the effects of a single dose of Mucinex ® 1200 mg (2 × 600 mg extended release tablets) (ER GGE) on 1) MCC/CC by assessing the rate of removal from the lung of inhaled radioactive tracer particles (Tc99m-sulfur colloid), 2) sputum dynamic rheology by stress/strain creep transformation over the linear part of the curve, 3) sessile drop interfacial tension by the deNouy ring technique, and 4) subjective symptom measures. MCC was measured during the morning (period 1) and compared to that in the afternoon 4 h later (period 2) immediately following either drug (n = 19) or placebo (n = 19). For both period 1 and 2 subjects performed 60 voluntary coughs from 60 to 90 min after inhalation of radio-labeled aerosol for a measure of CC. Sputum properties were measured from subjects who expectorated sputum during the cough period post treatment (n = 8–12 for each cohort). Results We found no effect of ER GGE on MCC or CC compared to placebo. MCC through 60 min for period 1 vs. 2 = 8.3 vs. 11.8% (placebo) and = 9.7 vs. 11.1% (drug) (NS) and CC for period 1 vs. 2 was 9.9 vs. 9.1% (placebo) and 10.8 vs. 5.6% (drug) (NS). There was no significant difference in sputum biophysical properties after administration of drug or placebo. Conclusions There was no significant effect of a single dose of ER GGE on MCC/CC or on sputum biophysical properties compared to placebo in this population of adult patients with an acute RTI. ClinicalTrials.gov Identifier: NCT01114581

The synthesis of Pt supported on zeolite 5A beads for the decarboxylation of oleic acid to heptadecane is demonstrated. The use of a microporous ZIF-67 crystalline layer on zeolite 5A beads not only improved the heptadecane selectivity but also, most importantly, improved the stability of the resultant catalyst. Heptadecane yields as high as ∼81% were observed for the fresh catalysts. The catalysts displayed only low to moderate loss of catalytic activity after two rounds of recycle. To our best knowledge, the catalytic performance of these catalysts is superior to those of the state-of-the-art catalysts at mild reaction conditions. In addition, as compared to powders, beads are much easier to recycle, can be fully recovered, and are more amenable for potential scale-up. The resultant catalysts are promising for the catalytic conversion of fatty acid molecules into gasoline/diesel-range hydrocarbons.

Chronic bronchitis (CB) is characterized by persistent cough and sputum production. Studies were performed to test whether mucus hyperconcentration and increased partial osmotic pressure, in part caused by abnormal purine nucleotide regulation of ion transport, contribute to the pathogenesis of CB.We tested the hypothesis that CB is characterized by mucus hyperconcentration, increased mucus partial osmotic pressures, and reduced mucus clearance.We measured in subjects with CB as compared with normal and asymptomatic smoking control subjects indices of mucus concentration (hydration; i.e., percentage solids) and sputum adenine nucleotide/nucleoside concentrations. In addition, sputum partial osmotic pressures and mucus transport rates were measured in subjects with CB.CB secretions were hyperconcentrated as indexed by an increase in percentage solids and total mucins, in part reflecting decreased extracellular nucleotide/nucleoside concentrations. CB mucus generated concentration-dependent increases in partial osmotic pressures into ranges predicted to reduce mucus transport. Mucociliary clearance (MCC) in subjects with CB was negatively correlated with mucus concentration (percentage solids). As a test of relationships between mucus concentration and disease, mucus concentrations and MCC were compared with FEV1, and both were significantly correlated.Abnormal regulation of airway surface hydration may slow MCC in CB and contribute to disease pathogenesis.

Heterogeneity of inhaled particle deposition in airways disease may be a sensitive indicator of physiologic changes in the lungs. Using planar gamma scintigraphy, we developed new methods to locate and quantify regions of high (hot) and low (cold) particle deposition in the lungs.Initial deposition and 24 hour retention images were obtained from healthy (n=31) adult subjects and patients with mild cystic fibrosis lung disease (CF) (n=14) following inhalation of radiolabeled particles (Tc99m-sulfur colloid, 5.4 μm MMAD) under controlled breathing conditions. The initial deposition image of the right lung was normalized to (i.e., same median pixel value), and then divided by, a transmission (Tc99m) image in the same individual to obtain a pixel-by-pixel ratio image. Hot spots were defined where pixel values in the deposition image were greater than 2X those of the transmission, and cold spots as pixels where the deposition image was less than 0.5X of the transmission. The number ratio (NR) of the hot and cold pixels to total lung pixels, and the sum ratio (SR) of total counts in hot pixels to total lung counts were compared between healthy and CF subjects. Other traditional measures of regional particle deposition, nC/P and skew of the pixel count histogram distribution, were also compared.The NR of cold spots was greater in mild CF, 0.221±0.047(CF) vs. 0.186±0.038 (healthy) (p0.005) and was significantly correlated with FEV1 %pred in the patients (R=-0.70). nC/P (central to peripheral count ratio), skew of the count histogram, and hot NR or SR were not different between the healthy and mild CF patients.These methods may provide more sensitive measures of airway function and localization of deposition that might be useful for assessing treatment efficacy in these patients.

Background Inhaled hypertonic saline (HS) has been shown to increase mucociliary clearance (MCC) and improve clinical outcomes in adults and adolescents with cystic fibrosis (CF). However, in younger children with CF, a large study failed to demonstrate clinical benefits. This discrepancy could reflect pharmacodynamic differences in the MCC response to HS in different populations. We previously demonstrated the absence of a sustained effect of HS on MCC in healthy adults and in this study sought to characterize the durability of the MCC response to HS in adults with CF. Methods At two study sites, MCC was measured in CF adults using gamma scintigraphy during three separate visits: at baseline, 15 min, and 4 h after a single dose of HS (7% NaCl, 4 mL). Particle clearance rates at these visits were used to assess the durability of the MCC response to HS. Results The average 90-minute clearance rate measured 4 h after HS was significantly increased (21.81% ± 12.8) when compared to baseline (13.77% ± 8.7, p = .048) and showed no apparent slowing relative to the rate measured 15 min after HS. While not all subjects responded to HS, the acute response strongly predicted the sustained effect in these subjects (r = 0.896, p Conclusions These results suggest that, in contrast to healthy adults, a single dose of HS has a prolonged effect on MCC in adults with CF, which lasts at least 4 h. This may explain its clinical efficacy in this population.

This chapter discusses the need for innovative health literacy solutions to combat extensive chronic disease prevalence and costs. The authors explore the intersection of chronic disease management and health literacy. They provide specific examples of successful health literacy interventions for managing several highly prevalent chronic diseases. This is followed by suggestions on pairing research and practice to support effective disease management programs. In addition, the authors discuss strategies for collection and dissemination of knowledge gained from collaborations between researchers and practitioners. They identify current challenges specific to disseminating information from the health literacy field and offer potential solutions. The chapter concludes with a brief look at future directions and organizational opportunities to integrate health literacy practices to address the need for effective chronic disease management.

It is difficult to identify reliable reference genes for transcriptomic analyses in biofluids such as saliva. This situation is particularly relevant for the newborn population, where rapid development is associated with dynamic changes in gene expression. Real-time gene expression monitoring holds great promise for elucidating disrupted pathways that result in morbidities unique to this population, such as retinopathy of prematurity, but its impact depends on identifying stable and consistently expressed genes across a wide range of gestational ages. We extracted total RNA from 400 neonatal saliva samples (postconceptional ages: 32 5/7 to 48 2/7 weeks), converted it to cDNA, and pre-amplified and analyzed it by qPCR for three commonly used reference genes, ACTB, GAPDH, and YWHAZ. Relative quantification was determined using the Δ Ct method. Data were analyzed as a whole and also stratified by age and sex. Descriptive statistics and homogeneity of variance were performed to identify optimal reference genes. Data analyzed from all ages and both sexes showed significant expression variation for ACTB, while GAPDH and YWHAZ showed greater stability. Male infants exhibited increased expression variation compared to females for ACTB, but neither GAPDH nor YWHAZ showed significant variance for either sex. We suggest that ACTB is an unreliable reference gene for the newborn population. Males showed significantly more variation in ACTB expression compared to females, which suggests a sex-specific developmental role for this biomarker. By contrast, GAPDH and YWHAZ were less variable and therefore preferable for use in neonates. Our findings may improve the use of reference genes for the RT-qPCR platform in the newborn over a wide range of gestational ages, thereby minimizing the likelihood of erroneous interpretation of gene expression during rapid growth, development, and differentiation.

Turner syndrome is a sex chromosome aneuploidy with characteristic malformations. Amniotic fluid, a complex biological material, could contribute to the understanding of Turner syndrome pathogenesis. In this pilot study, global gene expression analysis of cell-free RNA in amniotic fluid supernatant was utilized to identify specific genes/organ systems that may play a role in Turner syndrome pathophysiology. Cell-free RNA from amniotic fluid of five mid-trimester Turner syndrome fetuses and five euploid female fetuses matched for gestational age was extracted, amplified, and hybridized onto Affymetrix(®) U133 Plus 2.0 arrays. Significantly differentially regulated genes were identified using paired t tests. Biological interpretation was performed using Ingenuity Pathway Analysis and BioGPS gene expression atlas. There were 470 statistically significantly differentially expressed genes identified. They were widely distributed across the genome. XIST was significantly down-regulated (p < 0.0001); SHOX was not differentially expressed. One of the most highly represented organ systems was the hematologic/immune system, distinguishing the Turner syndrome transcriptome from other aneuploidies we previously studied. Manual curation of the differentially expressed gene list identified genes of possible pathologic significance, including NFATC3, IGFBP5, and LDLR. Transcriptomic differences in the amniotic fluid of Turner syndrome fetuses are due to genome-wide dysregulation. The hematologic/immune system differences may play a role in early-onset autoimmune dysfunction. Other genes identified with possible pathologic significance are associated with cardiac and skeletal systems, which are known to be affected in females with Turner syndrome. The discovery-driven approach described here may be useful in elucidating novel mechanisms of disease in Turner syndrome.

Objective The objective of this study was to understand the biological pathways involved in twin–twin transfusion syndrome (TTTS) by performing global gene expression analysis of amniotic fluid (AF) cell-free RNA. Methods A prospective whole transcriptome microarray study analyzing cell-free RNA in AF from TTTS recipient twins and singleton controls was carried out. Significantly differentially regulated genes in TTTS cases (N = 8) versus matched controls (N = 8) were identified and pathways analyses performed. Significant gene expression differences between stage II TTTS recipients (N = 5) and stage III TTTS recipients with abnormal Doppler measurements (N = 5) were also analyzed. Results Analysis of paired data from TTTS cases and controls revealed differential expression of 801 genes, which were significantly enriched for neurological disease and cardiovascular system pathways. We also identified cardiovascular genes and pathways associated with the presence of critically abnormal Doppler measurements in stage III TTTS recipients. Conclusions This study provides the first transcriptome-wide data on the impact of TTTS on fetal development. Our results show that gene expression involving neurological and cardiovascular pathways are altered in recipient fetuses prior to surgical treatment. This has relevance for the origins of long-term complications seen in survivors and for the development of future fetal biomarkers. © 2013 John Wiley & Sons, Ltd.

Quantification of particle deposition in the lung by gamma scintigraphy requires a reference image for location of regions of interest (ROIs) and normalization to lung thickness. In various laboratories, the reference image is made by a transmission scan ((57)Co or (99m)Tc) or gas ventilation scan ((133)Xe or (81)Kr). There has not been a direct comparison of measures from the two methods.We compared (99m)Tc transmission scans to (133)Xe equilibrium ventilation scans as reference images for 38 healthy subjects and 14 cystic fibrosis (CF) patients for their effects on measures of regional particle deposition: the central-to-peripheral ratio of lung counts (C/P); and ROI area versus forced vital capacity. Whole right lung ROI was based on either an isocontour threshold of three times the soft tissue transmission (TT) or a threshold of 20% of peak xenon ventilation counts (XV). We used a central ROI drawn to 50% of height and of width of the whole right lung ROI and placed along the left lung margin and centered vertically.In general, the correlation of normalized C/P (nC/P) between the two methods was strong. However, the value of nC/P was significantly smaller for the XV method than the TT method. Regression equations for the relationship of nC/P between the two methods were, for healthy subjects, y=0.75x+0.61, R(2)=0.64 using rectangular ROIs and y=0.76x+0.45, R(2)=0.66 using isocontour ROIs; and for CF patients, y=0.94x+0.46, R(2)=0.43 and y=0.85x+0.42, R(2)=0.41, respectively.(1) A transmission scan with an isocontour outline in combination with a rectangular central region to define the lung borders may be more useful than a ventilation scan. (2) Close correlation of nC/Ps measured by transmission or gas ventilation should allow confident comparison of values determined by the two methods.

A novel concept of utilizing nanoporous coatings as effective nanovalves on microporous adsorbents was developed for high capacity natural gas storage at low storage pressure. The work reported here for the first time presents the concept of nanovalved adsorbents capable of sealing high pressure CH4 inside the adsorbents and storing it at low pressure. Traditional natural gas storage tanks are thick and heavy, which makes them expensive to manufacture and highly energy-consuming to carry around. Our design uses unique adsorbent pellets with nanoscale pores surrounded by a coating that functions as a valve to help manage the pressure of the gas and facilitate more efficient storage and transportation. We expect this new concept will result in a lighter, more affordable product with increased storage capacity. The nanovalved adsorbent concept demonstrated here can be potentially extended for the storage of other important gas molecules targeted for diverse relevant functional applications.

It is now a well-established fact that fetal cells are present in the maternal circulation. The first evidence that fetal cells exist outside the fetoplacental unit was demonstrated in 1893 by Schmorl, who described the presence of trophoblast sprouts in the lungs of women who died of eclampsia (1). With the advent of sensitive molecular techniques, this finding has now been confirmed on numerous occasions. In 1969, Walknowska published a landmark study showing the detection of cells bearing a Y-chromosome in the blood of women carrying a male fetus (2). Since this report, the potential applications of using circulating fetal cells in prenatal screening have been researched extensively. Fetal cells represent a unique source of fetal DNA and RNA that can be used for the detection of fetal chromosomal aneuploidies and single gene disorders. In addition, new studies show that the fetoplacental unit is not as distinctly separate from the mother as was previously thought. In every pregnancy, there is trafficking of maternal and fetal cells and cell-free nucleic acids to and from the fetoplacental unit, respectively. Under normal circumstances, very few fetal cells enter the maternal bloodstream. However, in the case of many fetal or placental disorders, including aneuploidy or preeclampsia, an abnormally high number of fetal cells are present in the maternal circulation. Current data suggest that fetomaternal trafficking is a reflection not only of fetal health but also of placental development and pregnancy well-being. New evidence shows that the persistence of fetal cells in maternal tissues after delivery plays a role in subsequent maternal health. This chapter will discuss the role of circulating fetal cells in prenatal diagnosis and the relationship between abnormal fetomaternal trafficking of cells and the occurrence of disease in the woman both during gestation and postpartum.

Remodeling of adipose tissue is required to support the expansion of adipose mass. In obesity, an increased death of adipocytes contributes to the accelerated cellular turnover. We have shown that obesity in pregnancy is associated with metabolic and immune alterations in the adipose tissue. In this study, we characterized the mechanisms responsible for increased death of adipose cells of pregnant obese women and its functional consequences. We postulated that a higher turnover of dead cells in white adipose tissue of obese women would translate into release of cell-free DNA (cfDNA) into their systemic circulation. Increase in adipose mass of obese compared to lean women results from a lesser number of hypertrophic adipocytes and an accumulation of macrophages in the stromal vascular fraction (SVF). The adipocytes of obese displayed enhanced necrosis with a loss of perilipin staining at the plasma membrane. Apoptosis was prominent in SVF cells with an increased expression of caspase 9 and caspase 3 and a higher rate of terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end-labeling (TUNEL) positive CD68 macrophages in obese vs. lean. Whereas circulating fetal cfDNA concentrations were not changed, there was a twofold increase in circulating glyceraldehyde-3-phosphate dehydrogenase (GAPDH) cfDNA and adipose tissue GAPDH mRNA in obese women. The maternal systemic GAPDH cfDNA was positively correlated with BMI and gestational weight gain. These data suggest that the active remodeling of adipose tissue of obese pregnant women results in an increased release of cfDNA of maternal origin into the circulation.

Trisomies 18 and 21 are the two most common live born autosomal aneuploidies in humans. While the anatomic abnormalities in affected fetuses are well documented, the dysregulated biological pathways associated with the development of the aneuploid phenotype are less clear. Amniotic fluid (AF) cell-free RNA is a valuable source of biological information obtainable from live fetuses. In this study, we mined gene expression data previously produced by our group from mid-trimester AF supernatant samples. We identified the euploid, trisomy 18 and trisomy 21 AF transcriptomes, and analyzed them with a particular focus on the nervous system. We used multiple bioinformatics resources, including DAVID, Ingenuity Pathway Analysis, and the BioGPS Gene Expression Atlas. Our analyses confirmed that AF supernatant from aneuploid fetuses is enriched for nervous system gene expression and neurological disease pathways. Tissue analysis showed that fetal brain cortex and Cajal-Retzius cells were significantly enriched for genes contained in the AF transcriptomes. We also examined AF transcripts known to be dysregulated in aneuploid fetuses compared with euploid controls and identified several brain-specific transcripts among them. Many of these genes play critical roles in nervous system development. NEUROD2, which was downregulated in trisomy 18, induces neurogenic differentiation. SOX11, downregulated in trisomy 21, is a transcription factor that is essential for pan-neuronal protein expression and axonal growth of sensory neurons. Our results show that whole transcriptome analysis of cell-free RNA in AF from live pregnancies permits discovery of biomarkers of abnormal human neurodevelopment and advances our understanding of the pathophysiology of aneuploidy.

Amniotic fluid is a complex biological material that provides a unique window into the developing human. Residual amniotic fluid supernatant contains cell-free fetal RNA. The objective of this study was to develop an understanding of the amniotic fluid core transcriptome by analyzing the transcripts ubiquitously present in the amniotic fluid supernatant of euploid midtrimester fetuses.This was an in silico (computational) investigation using publicly available gene expression data previously produced by our group from 12 euploid midtrimester amniotic fluid samples. Functional analyses were performed using a web-based software analysis tool. Organ specificity was examined for each transcript using a gene expression atlas. For fetal organs not represented in the atlas, manual literature searching and the web-based software analysis tool were used to generate fetal organ-associated gene lists.There were 476 well-annotated genes present in 12 of 12 amniotic fluid samples. Functional analysis identified six physiologic systems represented in the amniotic fluid core transcriptome, including musculoskeletal and nervous system development and function and embryonic and organismal development. Mammalian target of rapamycin signaling was identified as a key canonical pathway. Twenty-three highly organ-specific transcripts were identified; six of these are known to be highly expressed in the fetal brain.Amniotic fluid cell-free fetal RNA can provide biological information on multiple fetal organ systems. The presence of fetal-brain specific transcripts in amniotic fluid suggests novel approaches to the study of developmental disorders that involve the central nervous system. The finding that the mammalian target of rapamycin signaling is enriched in midtrimester fetuses may have future applications in the study of fetal growth disorders.