Glycogen synthase kinase 3 activity enhances liver inflammation in MASH.

Background & Aims: Metabolic dysfunction-associated steatohepatitis (MASH) is characterized by excessive circulating toxic lipids, hepatic steatosis, and liver inflammation. Monocyte adhesion to liver sinusoidal endothelial cells (LSECs) and transendothelial migration (TEM) are crucial in the inflammatory process. Under lipotoxic stress, LSECs develop a proinflammatory phenotype known as endotheliopathy. However, mediators of endotheliopathy remain unclear. Methods: Primary mouse LSECs isolated from C57BL/6J mice fed chow or MASH-inducing diets rich in fat, fructose, and cholesterol (FFC) were subjected to multi-omics profiling. Mice with established MASH resulting from a choline-deficient high-fat diet (CDHFD) or FFC diet were also treated with two structurally distinct GSK3 inhibitors (LY2090314 and elraglusib [9-ING-41]). Results: Integrated pathway analysis of the mouse LSEC proteome and transcriptome indicated that leukocyte TEM and focal adhesion were the major pathways altered in MASH. Kinome profiling of the LSEC phosphoproteome identified glycogen synthase kinase (GSK)-3ß as the major kinase hub in MASH. GSK3ß-activating phosphorylation was increased in primary human LSECs treated with the toxic lipid palmitate and in human MASH. Palmitate upregulated the expression of C-X-C motif chemokine ligand 2, intracellular adhesion molecule 1, and phosphorylated focal adhesion kinase, via a GSK3-dependent mechanism. Congruently, the adhesive and transendothelial migratory capacities of primary human neutrophils and THP-1 monocytes through the LSEC monolayer under lipotoxic stress were reduced by GSK3 inhibition. Treatment with the GSK3 inhibitors LY2090314 and elraglusib ameliorated liver inflammation, injury, and fibrosis in FFC- and CDHFD-fed mice, respectively. Immunophenotyping using cytometry by mass cytometry by time of flight of intrahepatic leukocytes from CDHFD-fed mice treated with elraglusib showed reduced infiltration of proinflammatory monocyte-derived macrophages and monocyte-derived dendritic cells. Conclusion: GSK3 inhibition attenuates lipotoxicity-induced LSEC endotheliopathy and could serve as a potential therapeutic strategy for treating human MASH. Impact and Implications: LSECs under lipotoxic stress in MASH develop a proinflammatory phenotype known as endotheliopathy, with obscure mediators and functional outcomes. The current study identified GSK3 as the major driver of LSEC endotheliopathy, examined its pathogenic role in myeloid cell-associated liver inflammation, and defined the therapeutic efficacy of pharmacological GSK3 inhibitors in murine MASH. This study provides preclinical data for the future investigation of GSK3 pharmacological inhibitors in human MASH. The results of this study are important to hepatologists, vascular biologists, and investigators studying the mechanisms of inflammatory liver disease and MASH, as well as those interested in drug development.

Preterm and Early-Term Delivery After Heat Waves in 50 US Metropolitan Areas.

Importance: Heat waves are increasing in frequency, intensity, and duration and may be acutely associated with pregnancy outcomes. Objective: To examine changes in daily rates of preterm and early-term birth after heat waves in a 25-year nationwide study. Design, Setting, and Participants: This cohort study of singleton births used birth records from 1993 to 2017 from the 50 most populous US metropolitan statistical areas (MSAs). The study included 53 million births, covering 52.8% of US births over the period. Data were analyzed between October 2022 and March 2023 at the National Center for Health Statistics. Exposures: Daily temperature data from Daymet at 1-km2 resolution were averaged over each MSA using population weighting. Heat waves were defined in the 4 days (lag, 0-3 days) or 7 days (lag, 0-6 days) preceding birth. Main Outcomes and Measures: Daily counts of preterm birth (28 to <37 weeks), early-term birth (37 to <39 weeks), and ongoing pregnancies in each gestational week on each day were enumerated in each MSA. Rate ratios for heat wave metrics were obtained from time-series models restricted to the warm season (May to September) adjusting for MSA, year, day of season, and day of week, and offset by pregnancies at risk. Results: There were 53 154 816 eligible births in the 50 MSAs from 1993 to 2017; 2 153 609 preterm births and 5 795 313 early-term births occurring in the warm season were analyzed. A total of 30.0% of mothers were younger than 25 years, 53.8% were 25 to 34 years, and 16.3% were 35 years or older. Heat waves were positively associated with daily rates of preterm and early-term births, showing a dose-response association with heat wave duration and temperatures and stronger associations in the more acute 4-day window. After 4 consecutive days of mean temperatures exceeding the local 97.5th percentile, the rate ratio for preterm birth was 1.02 (95% CI, 1.00-1.03), and the rate ratio for early-term birth was 1.01 (95% CI, 1.01-1.02). For the same exposure, among those who were 29 years of age or younger, had a high school education or less, and belonged to a racial or ethnic minority group, the rate ratios were 1.04 (95% CI, 1.02-1.06) for preterm birth and 1.03 (95% CI, 1.02-1.05) for early-term birth. Results were robust to alternative heat wave definitions, excluding medically induced deliveries, and alternative statistical model specifications. Conclusions and Relevance: In this cohort study, preterm and early-term birth rates increased after heat waves, particularly among socioeconomically disadvantaged subgroups. Extreme heat events have implications for perinatal health.

A multiscale analysis of heatwaves and urban heat islands in the western U.S. during the summer of 2021.

Extreme heat events are occurring more frequently and with greater intensity due to climate change. They result in increased heat stress to populations causing human health impacts and heat-related deaths. The urban environment can also exacerbate heat stress because of man-made materials and increased population density. Here we investigate the extreme heatwaves in the western U.S. during the summer of 2021. We show the atmospheric scale interactions and spatiotemporal dynamics that contribute to increased temperatures across the region for both urban and rural environments. In 2021, daytime maximum temperatures during heat events in eight major cities were 10-20 °C higher than the 10-year average maximum temperature. We discuss the temperature impacts associated with processes across scales: climate or long-term change, the El Niño-Southern Oscillation, synoptic high-pressure systems, mesoscale ocean/lake breezes, and urban climate (i.e., urban heat islands). Our findings demonstrate the importance of scale interactions impacting extreme heat and the need for holistic approaches in heat mitigation strategies.

Utility of new image-derived biomarkers for autosomal dominant polycystic kidney disease prognosis using automated instance cyst segmentation.

New image-derived biomarkers for patients affected by autosomal dominant polycystic kidney disease are needed to improve current clinical management. The measurement of total kidney volume (TKV) provides critical information for clinicians to drive care decisions. However, patients with similar TKV may present with very different phenotypes, often requiring subjective decisions based on other factors (e.g., appearance of healthy kidney parenchyma, a few cysts contributing significantly to overall TKV, etc.). In this study, we describe a new technique to individually segment cysts and quantify biometric parameters including cyst volume, cyst number, parenchyma volume, and cyst parenchyma surface area. Using data from the Consortium for Radiologic Imaging Studies of Polycystic Kidney Disease (CRISP) study the utility of these new parameters was explored, both quantitatively as well as visually. Total cyst number and cyst parenchyma surface area showed superior prediction of the slope of estimated glomerular filtration rate decline, kidney failure and chronic kidney disease stages 3A, 3B, and 4, compared to TKV. In addition, presentations such as a few large cysts contributing significantly to overall kidney volume were shown to be much better stratified in terms of outcome predictions. Thus, these new image biomarkers, which can be obtained automatically, will have great utility in future studies and clinical care for patients affected by autosomal dominant polycystic kidney disease.

Statistical Comparison and Assessment of Four Fire Emissions Inventories for 2013 and a Large Wildfire in the Western United States.

Wildland fires produce smoke plumes that impact air quality and human health. To understand the effects of wildland fire smoke on humans, the amount and composition of the smoke plume must be quantified. Using a fire emissions inventory is one way to determine the emissions rate and composition of smoke plumes from individual fires. There are multiple fire emissions inventories, and each uses a different method to estimate emissions. This paper presents a comparison of four emissions inventories and their products: Fire INventory from NCAR (FINN version 1.5), Global Fire Emissions Database (GFED version 4s), Missoula Fire Labs Emissions Inventory (MFLEI (250 m) and MFLEI (10 km) products), and Wildland Fire Emissions Inventory System (WFEIS (MODIS) and WFEIS (MTBS) products). The outputs from these inventories are compared directly. Because there are no validation datasets for fire emissions, the outlying points from the Bayesian models developed for each inventory were compared with visible images and fire radiative power (FRP) data from satellite remote sensing. This comparison provides a framework to check fire emissions inventory data against additional data by providing a set of days to investigate closely. Results indicate that FINN and GFED likely underestimate emissions, while the MFLEI products likely overestimate emissions. No fire emissions inventory matched the temporal distribution of emissions from an external FRP dataset. A discussion of the differences impacting the emissions estimates from the four fire emissions inventories is provided, including a qualitative comparison of the methods and inputs used by each inventory and the associated strengths and limitations.

Sequence analysis and function of mosquito aeCCC2 and Drosophila Ncc83 orthologs.

Dipteran insects have genes that code for two different Na+-dependent cation-chloride cotransporter (CCC) paralogs. Aedes aegypti aeNKCC1 is an ortholog of Drosophila melanogaster Ncc69, a bumetanide-sensitive Na+-K+-2Cl- cotransporter (NKCC). Aedes aegypti aeCCC2 and aeCCC3 are orthologs of Drosophila Ncc83. Prior work suggests that the transport properties of aeCCC2 differ from canonical NKCCs. In particular, Xenopus oocytes expressing aeCCC2 have increased Na+-dependent membrane currents compared to controls, whereas NKCCs are electroneutral. Here, we further evaluated the function and localization of aeCCC2 and Ncc83. In oocytes expressing aeCCC2 or Ncc83, membrane potential (Vm) hyperpolarized upon Na+ removal; following hypotonic exposure the change in Vm was greater than it was in controls. In voltage-clamp experiments, membrane currents were concentration dependent on Na+ with an apparent affinity (Km) of approximately 4.6 mM. In Malpighian tubules of larval and adult mosquitoes, aeCCC2 was localized along the basolateral aspect of principal cells. Sequence comparisons among transporters from Drosophila, Aedes, Anopheles, and Culex revealed 33 residues within the transmembrane domains (TMDs) that are fully conserved within paralogs but that differ between orthologs of NKCC1 and orthologs of aeCCC2/Ncc83. These residues are distributed across all 12 TMDs. Our results provide a foundation for further exploration of the structural basis for functional differences between insect Na+-dependent CCCs.

Estimating Heat-Related Exposures and Urban Heat Island Impacts: A Case Study for the 2012 Chicago Heatwave.

Accelerated urbanization increases both the frequency and intensity of heatwaves (HW) and urban heat islands (UHIs). An extreme HW event occurred in 2012 summer that caused temperatures of more than 40°C in Chicago, Illinois, USA, which is a highly urbanized city impacted by UHIs. In this study, multiple numerical models, including the High Resolution Land Data Assimilation System (HRLDAS) and Weather Research and Forecasting (WRF) model, were used to simulate the HW and UHI, and their performance was evaluated. In addition, sensitivity testing of three different WRF configurations was done to determine the impact of increasing model complexity in simulating urban meteorology. Model performances were evaluated based on the statistical performance metrics, the application of a multi-layer urban canopy model (MLUCM) helps WRF to provide the best performance in this study. HW caused rural temperatures to increase by ∼4°C, whereas urban Chicago had lower magnitude increases from the HW (∼2-3°C increases). Nighttime UHI intensity (UHII) ranged from 1.44 to 2.83°C during the study period. Spatiotemporal temperature fields were used to estimate the potential heat-related exposure and to quantify the Excessive Heat Factor (EHF). The EHF during the HW episode provides a risk map indicating that while urban Chicago had higher heat-related stress during this event, the rural area also had high risk, especially during nighttime in central Illinois. This study provides a reliable method to estimate spatiotemporal exposures for future studies of heat-related health impacts.

Confronting Uncertainties of Simulated Air Pollution Concentrations during Persistent Cold Air Pool Events in the Salt Lake Valley, Utah.

Air pollutant accumulations during wintertime persistent cold air pool (PCAP) events in mountain valleys are of great concern for public health worldwide. Uncertainties associated with the simulated meteorology under stable conditions over complex terrain hinder realistic simulations of air quality using chemical transport models. We use the Community Multiscale Air Quality (CMAQ) model to simulate the gaseous and particulate species for 1 month in January 2011 during the Persistent Cold Air Pool Study (PCAPS) in the Salt Lake Valley (SLV), Utah (USA). Results indicate that the temporal variability associated with the elevated NOx and PM2.5 concentrations during PCAP events was captured by the model (r = 0.20 for NOx and r = 0.49 for PM2.5). However, concentrations were not at the correct magnitude (NMB = -35/12% for PM2.5 during PCAPs/non-PCAPs), where PM2.5 was underestimated during PCAP events and overestimated during non-PCAP periods. The underestimated PCAP strength is represented by valley heat deficit, which contributed to the underestimated PM2.5 concentrations compared with observations due to the model simulating more vertical mixing and less stable stratification than what was observed. Based on the observations, the dominant PM2.5 species were ammonium and nitrate. We provide a discussion that aims to investigate the emissions and chemistry model uncertainties using the nitrogen ratio method and the thermodynamic ammonium nitrate regime method.

Electrogenic sodium bicarbonate cotransporter NBCe1 regulates pancreatic ß cell function in type 2 diabetes.

Pancreatic ß cell failure in type 2 diabetes mellitus (T2DM) is attributed to perturbations of the ß cell's transcriptional landscape resulting in impaired glucose-stimulated insulin secretion. Recent studies identified SLC4A4 (a gene encoding an electrogenic Na+-coupled HCO3- cotransporter and intracellular pH regulator, NBCe1) as one of the misexpressed genes in ß cells of patients with T2DM. Thus, in the current study, we set out to test the hypothesis that misexpression of SLC4A4/NBCe1 in T2DM ß cells contributes to ß cell dysfunction and impaired glucose homeostasis. To address this hypothesis, we first confirmed induction of SLC4A4/NBCe1 expression in ß cells of patients with T2DM and demonstrated that its expression was associated with loss of ß cell transcriptional identity, intracellular alkalinization, and ß cell dysfunction. In addition, we generated a ß cell-selective Slc4a4/NBCe1-KO mouse model and found that these mice were protected from diet-induced metabolic stress and ß cell dysfunction. Importantly, improved glucose tolerance and enhanced ß cell function in Slc4a4/NBCe1-deficient mice were due to augmented mitochondrial function and increased expression of genes regulating ß cell identity and function. These results suggest that increased ß cell expression of SLC4A4/NBCe1 in T2DM plays a contributory role in promotion of ß cell failure and should be considered as a potential therapeutic target.

Use of 3D Robotic Ultrasound for In Vivo Analysis of Mouse Kidneys.

Common modalities for in vivo imaging of rodents include positron emission tomography (PET), computed tomography (CT), magnetic resonance imaging (MRI), and ultrasound (US). Each method has limitations and advantages, including availability, ease of use, cost, size, and the use of ionizing radiation or magnetic fields. This protocol describes the use of 3D robotic US for in vivo imaging of rodent kidneys and heart, subsequent data analysis, and possible research applications. Practical applications of robotic US are the quantification of total kidney volume (TKV), as well as the measurement of cysts, tumors, and vasculature. Although the resolution is not as high as other modalities, robotic US allows for more practical high throughput data collection. Furthermore, using US M-mode imaging, cardiac function may be quantified. Since the kidneys receive 20%-25% of the cardiac output, assessing cardiac function is critical to the understanding of kidney physiology and pathophysiology.

Lithium heparin interference in the Abbott enzymatic creatinine assay: the significance of under-filled tubes.

BACKGROUND: Spuriously high results using the Abbott Architect enzymatic creatinine assay were noted to be particularly associated with very small sample volumes. This led us to query the effect of under-filling lithium heparin tubes on the measured enzymatic creatinine result. METHODS: Blood was provided by 5 laboratory personnel and then decanted into 5 x1.2 mL Sarstedt S-Monovette tubes, giving final blood volumes of 200, 400, 600, 800 and 1200 µL. Plasma was analysed using Abbott Architect Jaffe, enzymatic creatinine, Beckman Coulter (AU500) enzymatic creatinine and Roche (Cobas c702) enzymatic creatinine assays. Saline was also added to Sarstedt 1.2 mL and Teklab 2 mL tubes and analysed using the Abbott Jaffe and enzymatic creatinine methods. RESULTS: Increasing degrees of under-fill were associated with greater over-estimation of creatinine using the Abbott enzymatic assay, but no difference was noted using Jaffe methodology on the same platform or enzymatic assays provided by Roche or Beckman. On average, creatinine was 40.6% (+27.7 µmol/L) higher when only 200 µL of blood was present in the tube. Small volumes of saline added to lithium heparin tubes measured significant creatinine concentrations using the Abbott enzymatic method. CONCLUSIONS: Lithium heparin directly interferes in the Abbott Architect enzymatic creatinine assay. Under-filling lithium heparin tubes can lead to clinically significant over-estimation of creatinine results by this assay. Users of this assay should be aware of the potential for spurious results in small sample volumes collected into lithium heparin tubes and implement robust procedures for identifying and reporting results on these samples.

Coupled Air Quality and Boundary-Layer Meteorology in Western U.S. Basins during Winter: Design and Rationale for a Comprehensive Study.

Wintertime episodes of high aerosol concentrations occur frequently in urban and agricultural basins and valleys worldwide. These episodes often arise following development of persistent cold-air pools (PCAPs) that limit mixing and modify chemistry. While field campaigns targeting either basin meteorology or wintertime pollution chemistry have been conducted, coupling between interconnected chemical and meteorological processes remains an insufficiently studied research area. Gaps in understanding the coupled chemical-meteorological interactions that drive high pollution events make identification of the most effective air-basin specific emission control strategies challenging. To address this, a September 2019 workshop occurred with the goal of planning a future research campaign to investigate air quality in Western U.S. basins. Approximately 120 people participated, representing 50 institutions and 5 countries. Workshop participants outlined the rationale and design for a comprehensive wintertime study that would couple atmospheric chemistry and boundary-layer and complex-terrain meteorology within western U.S. basins. Participants concluded the study should focus on two regions with contrasting aerosol chemistry: three populated valleys within Utah (Salt Lake, Utah, and Cache Valleys) and the San Joaquin Valley in California. This paper describes the scientific rationale for a campaign that will acquire chemical and meteorological datasets using airborne platforms with extensive range, coupled to surface-based measurements focusing on sampling within the near-surface boundary layer, and transport and mixing processes within this layer, with high vertical resolution at a number of representative sites. No prior wintertime basin-focused campaign has provided the breadth of observations necessary to characterize the meteorological-chemical linkages outlined here, nor to validate complex processes within coupled atmosphere-chemistry models.

Acute associations between heatwaves and preterm and early-term birth in 50 US metropolitan areas: a matched case-control study.

BACKGROUND: The effect of heatwaves on adverse birth outcomes is not well understood and may vary by how heatwaves are defined. The study aims to examine acute associations between various heatwave definitions and preterm and early-term birth. METHODS: Using national vital records from 50 metropolitan statistical areas (MSAs) between 1982 and 1988, singleton preterm (< 37 weeks) and early-term births (37-38 weeks) were matched (1:1) to controls who completed at least 37 weeks or 39 weeks of gestation, respectively. Matching variables were MSA, maternal race, and maternal education. Sixty heatwave definitions including binary indicators for exposure to sustained heat, number of high heat days, and measures of heat intensity (the average degrees over the threshold in the past 7 days) based on the 97.5th percentile of MSA-specific temperature metrics, or the 85th percentile of positive excessive heat factor (EHF) were created. Odds ratios (OR) for heatwave exposures in the week preceding birth (or corresponding gestational week for controls) were estimated using conditional logistic regression adjusting for maternal age, marital status, and seasonality. Effect modification by maternal education, age, race/ethnicity, child sex, and region was assessed. RESULTS: There were 615,329 preterm and 1,005,576 early-term case-control pairs in the analyses. For most definitions, exposure to heatwaves in the week before delivery was consistently associated with increased odds of early-term birth. Exposure to more high heat days and more degrees above the threshold yielded higher magnitude ORs. For exposure to 3 or more days over the 97.5th percentile of mean temperature in the past week compared to zero days, the OR was 1.027 for early-term birth (95%CI: 1.014, 1.039). Although we generally found null associations when assessing various heatwave definitions and preterm birth, ORs for both preterm and early-term birth were greater in magnitude among Hispanic and non-Hispanic black mothers. CONCLUSION: Although associations varied across metrics and heatwave definitions, heatwaves were more consistently associated with early-term birth than with preterm birth. This study's findings may have implications for prevention programs targeting vulnerable subgroups as climate change progresses.

Expression of the regulated isoform of the electrogenic Na+/HCO3- cotransporter, NBCe1, is enriched in pacemaker interstitial cells of Cajal.

Interstitial cells of Cajal (ICCs) generate electrical slow waves, which are required for normal gastrointestinal motility. The mechanisms for generation of normal pacemaking are not fully understood. Normal gastrointestinal contractility- and electrical slow-wave activity depend on the presence of extracellular HCO3-. Previous transcriptional analysis identified enrichment of mRNA encoding the electrogenic Na+/HCO3- cotransporter (NBCe1) gene (Slc4a4) in pacemaker myenteric ICCs in mouse small intestine. We aimed to determine the distribution of NBCe1 protein in ICCs of the mouse gastrointestinal tract and to identify the transcripts of the Slc4a4 gene in mouse and human small intestinal tunica muscularis. We determined the distribution of NBCe1 immunoreactivity (NBCe1-IR) by immunofluorescent labeling in mouse and human tissues. In mice, NBCe1-IR was restricted to Kit-positive myenteric ICCs of the stomach and small intestine and submuscular ICCs of the large intestine, that is, the slow wave generating subset of ICCs. Other subtypes of ICCs were NBCe1-negative. Quantitative real-time PCR identified >500-fold enrichment of Slc4a4-207 and Slc4a4-208 transcripts ["IP3-receptor-binding protein released by IP3" (IRBIT)-regulated isoforms] in Kit-expressing cells isolated from KitcreERT2/+, Rpl22tm1.1Psam/Sj mice and from single GFP-positive ICCs from Kittm1Rosay mice. Human jejunal tunica muscularis ICCs were also NBCe1-positive, and SLC4A4-201 and SLC4A4-204 RNAs were >300-fold enriched relative to SLC4A4-202. In summary, NBCe1 protein expressed in ICCs with electrical pacemaker function is encoded by Slc4a4 gene transcripts that generate IRBIT-regulated isoforms of NBCe1. In conclusion, Na+/HCO3- cotransport through NBCe1 contributes to the generation of pacemaker activity in subsets of ICCs.NEW & NOTEWORTHY In this study, we show that the electrogenic Na+/HCO3- cotransporter, NBCe1/Slc4a4, is expressed in subtypes of interstitial cells of Cajal (ICCs) responsible for electrical slow wave generation throughout the mouse gastrointestinal tract and is absent in other types of ICCs. The transcripts of Slc4a4 expressed in mouse ICCs and human gastrointestinal smooth muscle are the regulated isoforms. This indicates a key role for HCO3- transport in generation of gastrointestinal motility patterns.

Spontaneous pneumomediastinum: an unusual complication of hyperemesis gravidarum.

A 21-year-old first trimester primagravida woman with hyperemesis gravidarum was noted to have incidental subcutaneous emphysema during thyroid ultrasound. Follow-up radiograph demonstrated supraclavicular subcutaneous emphysema, left apical pneumothorax and pneumomediastinum. The patient was transferred to the intensive care unit and evaluated for esophageal rupture. Because no defects were seen on swallow studies/endoscopy and the patient was stable, she was treated conservatively with antibiotics and monitored. The patient's condition improved, and she was discharged on hospital day 6. Subcutaneous emphysema secondary to hyperemesis gravidarum is a rare but potentially life-threatening condition in which the source of the mediastinal leak needs to be immediately determined. After the more serious sources of mediastinal air have been excluded, conservative management is suitable.

Specialized stellate cells offer a privileged route for rapid water flux in Drosophila renal tubule.

Insects are highly successful, in part through an excellent ability to osmoregulate. The renal (Malpighian) tubules can secrete fluid faster on a per-cell basis than any other epithelium, but the route for these remarkable water fluxes has not been established. In Drosophila melanogaster, we show that 4 genes of the major intrinsic protein family are expressed at a very high level in the fly renal tissue: the aquaporins (AQPs) Drip and Prip and the aquaglyceroporins Eglp2 and Eglp4 As predicted from their structure, and by their transport function by expressing these proteins in Xenopus oocytes, Drip, Prip, and Eglp2 show significant and specific water permeability, whereas Eglp2 and Eglp4 show very high permeability to glycerol and urea. Knockdowns of any of these genes result in impaired hormone-induced fluid secretion. The Drosophila tubule has 2 main secretory cell types: active cation-transporting principal cells, wherein the aquaglyceroporins localize to opposite plasma membranes, and small stellate cells, the site of the chloride shunt conductance, with these AQPs localizing to opposite plasma membranes. This suggests a model in which osmotically obliged water flows through the stellate cells. Consistent with this model, fluorescently labeled dextran, an in vivo marker of membrane water permeability, is trapped in the basal infoldings of the stellate cells after kinin diuretic peptide stimulation, confirming that these cells provide the major route for transepithelial water flux. The spatial segregation of these components of epithelial water transport may help to explain the unique success of the higher insects in regulating their internal environments.

Assessing Polycystic Kidney Disease in Rodents: Comparison of Robotic 3D Ultrasound and Magnetic Resonance Imaging.

Polycystic kidney disease (PKD) is an inherited disorder characterized by renal cyst formation and enlargement of the kidney. PKD severity can be staged noninvasively by measuring total kidney volume (TKV), a promising biomarker that has recently received regulatory qualification. In preclinical mouse models, where the disease is studied and potential therapeutics are evaluated, the most popular noninvasive method of measuring TKV is magnetic resonance imaging (MRI). Although MRI provides excellent 3D resolution and contrast, these systems are expensive to operate, have long acquisition times, and, consequently, are not heavily used in preclinical PKD research. In this study, a new imaging instrument, based on robotic ultrasound (US), was evaluated as a complementary approach for assessing PKD in rodent models. The objective was to determine the extent to which TKV measurements on the robotic US scanner correlated with both in vivo and ex vivo reference standards (MRI and Vernier calipers, respectively). A cross-sectional study design was implemented that included both PKD-affected mice and healthy wild types, spanning sex and age for a wide range of kidney volumes. It was found that US-derived TKV measurements and kidney lengths were strongly associated with both in vivo MRI and ex vivo Vernier caliper measurements (R 2=0.94 and 0.90, respectively). In addition to measuring TKV, renal vascular density was assessed using acoustic angiography (AA), a novel contrast-enhanced US methodology. AA image intensity, indicative of volumetric vascularity, was seen to have a strong negative correlation with TKV (R 2=0.82), suggesting impaired renal vascular function in mice with larger kidneys. These studies demonstrate that robotic US can provide a rapid and accurate approach for noninvasively evaluating PKD in rodent models.

Source apportionment of primary and secondary PM2.5: Associations with pediatric respiratory disease emergency department visits in the U.S. State of Georgia.

We developed a hybrid chemical transport model and receptor model (CTM-RM) to conduct source apportionment of both primary and secondary PM2.5 (particulate matter ≤2.5 µm in diameter) at 36 km resolution throughout the U.S. State of Georgia for the years 2005 and 2007. This novel source apportionment model enabled us to estimate and compare associations of short-term changes in 12 PM2.5 source concentrations (agriculture, biogenic, coal, dust, fuel oil, metals, natural gas, non-road mobile diesel, non-road mobile gasoline, on-road mobile diesel, on-road mobile gasoline, and all other sources) with emergency department (ED) visits for pediatric respiratory diseases. ED visits for asthma (N = 49,651), pneumonia (N = 25,558), and acute upper respiratory infections (acute URI, N = 235,343) among patients aged ≤18 years were obtained from patient claims records. Using a case-crossover study, we estimated odds ratios per interquartile range (IQR) increase for 3-day moving average PM2.5 source concentrations using conditional logistic regression, matching on day-of-week, month, and year, and adjusting for average temperature, humidity, and holidays. We fit both single-source and multi-source models. We observed positive associations between several PM2.5 sources and ED visits for asthma, pneumonia, and acute URI. For example, for asthma, per IQR increase in the source contribution in the single-source model, odds ratios were 1.022 (95% CI: 1.013, 1.031) for dust; 1.050 (95% CI: 1.036, 1.063) for metals, and 1.091 (95% CI: 1.064, 1.119) for natural gas. These sources comprised 5.7%, 2.2%, and 6.3% of total PM2.5 mass, respectively. PM2.5 from metals and natural gas were positively associated with all three respiratory outcomes. In addition, non-road mobile diesel was positively associated with pneumonia and acute URI.

A Bayesian ensemble approach to combine PM2.5 estimates from statistical models using satellite imagery and numerical model simulation.

Ambient fine particulate matter less than 2.5 µm in aerodynamic diameter (PM2.5) has been linked to various adverse health outcomes. PM2.5 arises from both natural and anthropogenic sources, and PM2.5 concentrations can vary over space and time. However, the sparsity of existing air quality monitors greatly restricts the spatial-temporal coverage of PM2.5 measurements, potentially limiting the accuracy of PM2.5-related health studies. Various methods exist to address these limitations by supplementing air quality monitoring measurements with additional data. We develop a method to combine PM2.5 estimated from satellite-retrieved aerosol optical depth (AOD) and chemical transport model (CTM) simulations using statistical models. While most previous methods utilize AOD or CTM separately, we aim to leverage advantages offered by both data sources in terms of resolution and coverage using Bayesian ensemble averaging. Our approach differs from previous ensemble approaches in its ability to not only incorporate uncertainties in PM2.5 estimates from individual models but also to provide uncertainties for the resulting ensemble estimates. In an application of estimating daily PM2.5 in the Southeastern US, the ensemble approach outperforms previously developed spatial-temporal statistical models that use either AOD or bias-corrected CTM simulations in cross-validation (CV) analyses. More specifically, in spatially clustered CV experiments, the ensemble approach reduced the AOD-only and CTM-only model's root mean squared error (RMSE) by at least 13%. Similar improvements were seen in R2. The enhanced prediction performance that the ensemble technique provides at fine-scale spatial resolution, as well as the availability of prediction uncertainty, can be further used in health effect analyses of air pollution exposure.