Impacts of phosphorus amendments on legacy soil contamination from lead-based paint on a California, USA university campus.

Lead (Pb) is one of the most common heavy metal urban soil contaminants with well-known toxicity to humans. This incubation study (2-159 d) compared the ability of bone meal (BM), potassium hydrogen phosphate (KP), and triple superphosphate (TSP), at phosphorus:lead (P:Pb) molar ratios of 7.5:1, 15:1, and 22.5:1, to reduce bioaccessible Pb in soil contaminated by Pb-based paint relative to control soil to which no P amendment was added. Soil pH and Mehlich 3 bioaccessible Pb and P were measured as a function of incubation time and amount and type of P amendment. XAS assessed Pb speciation after 30 and 159 d of incubation. The greatest reductions in bioaccessible Pb at 159 d were measured for TSP at the 7.5:1 and 15:1 P:Pb molar ratios. The 7.5:1 KP treatment was the only other treatment with significant reductions in bioaccessible Pb compared to the control soil. It is unclear why greater reductions of bioaccessible Pb occurred with lower P additions, but it strongly suggests that the amount of P added was not a controlling factor in reducing bioaccessible Pb. This was further supported because Pb-phosphates were not detected in any samples using XAS. The most notable difference in the effect of TSP versus other amendments was the reduction in pH. However, the relationship between increasing TSP additions, resulting in decreasing pH and decreasing Pb bioaccessibility was not consistent. The 22.5:1 P:Pb TSP treatment had the lowest pH but did not significantly reduce bioaccessible Pb compared to the control soil. The 7.5:1 and 15:1 P:Pb TSP treatments significantly reduced bioaccessible Pb relative to the control and had significantly higher pH than the 22.5:1 P:Pb treatment. Clearly, impacts of P additions and soil pH on Pb bioaccessibility require further investigation to decipher mechanisms governing Pb speciation in Pb-based paint contaminated soils.

Effect of the COVID-19 pandemic on racial disparities in postpartum visits at a Texas level IV maternal center.

Background: The primary aim of our study was to determine the attendance of postpartum visits stratified by race and if the COVID-19 pandemic affected racial disparities in postpartum visit attendance. Methods: We searched our labor and delivery records from July 1, 2019 to December 31, 2019 and from July 1, 2020 to December 31, 2020 and included patients who delivered liveborn infants. The final analysis was restricted to patients who identified as White or Caucasian only, Black or African American only, or Hispanic. We then performed joint tests on the logistic regression with an interaction term of race and year of delivery to determine the final model. Results: The odds ratio of Black or African American and Hispanic patients attending a postpartum visit was 0.589 (95% CI 0.456, 0.760; P < .001) and 0.836 (95% CI 0.676, 1.034; P = 0.099), respectively, compared to White or Caucasian patients. The interaction term of race and year of delivery was not statistically significant. Conclusion: Black or African American patients at our hospital had a clinically and statistically significant lower utilization of postpartum visits compared to White or Caucasian patients and this disparity was not exacerbated by the COVID-19 pandemic.

Effect of the COVID-19 pandemic on racial disparities in neuraxial labor analgesia at a Texas level IV maternal care center.

We hypothesized that racial disparities in labor epidural analgesia at our hospital that existed prior to the COVID-19 pandemic would be exacerbated during the COVID-19 pandemic. We examined patients who delivered vaginally at our hospital for the last 6 months of 2019 and the last 6 months of 2020. We performed joint testing of coefficient P values, and the interaction term between race and year of delivery was not significant (0.364). A multivariate logistic regression model found that Hispanic patients (odds ratio 0.555 [0.408, 0.756], P < 0.001) and Black or African American patients (odds ratio 0.613 [0.408, 0.921], P = 0.018) were less likely to receive labor epidural analgesia compared to White or Caucasian patients. Odds ratios of receiving labor epidural analgesia were higher with increasing gestational age (1.116 [1.067, 1.168], P < 0.001) and lower with increasing parity (0.789 [0.719, 0.867], P < 0.001). The year of birth that corresponded to before or during the COVID-19 pandemic did not predict whether a patient received labor epidural analgesia (1.247 [0.941, 1.652], P = 0.124). Because the interaction between race and year of birth was not statistically significant, we conclude that the COVID-19 pandemic did not exacerbate racial disparities in labor epidural analgesia at our hospital.

Choice of anesthetic technique for dilation and curettage for indication of pregnancy loss.

We hypothesized that patients at our hospital who received general anesthesia as the initial anesthetic technique for dilation and curettage for loss of pregnancy during the first or second trimesters would have a higher estimated blood loss compared to patients who had sedation. We searched our electronic medical record system for patients who had a dilation and curettage for the indication of loss of pregnancy during the first or second trimesters from July 1, 2018, to June 30, 2021. A total of 165 (72%) and 64 (28%) patients had general anesthesia and sedation, respectively, as the initial anesthetic technique. Patients who had general anesthesia and sedation had estimated blood loss interquartile ranges of 50 to 500 mL and 30 to 100 mL, respectively (P < 0.01). A multivariate model that controlled for gestational age and location of procedure found that the odds ratio of patients receiving sedation for dilation and curettage in the labor and delivery suite was 7.24 (95% confidence interval 2.92, 17.94; P < 0.01) compared to the main operating room. Dilation and curettage that used sedation was associated with a lower estimated blood loss and was more likely to be performed in the labor and delivery suite.

Anti-nociceptive efficacy of the soluble epoxide hydrolase inhibitor t-TUCB in horses with mechanically induced lameness.

INTRODUCTION: Soluble epoxide hydrolase (sEH) inhibitors are novel anti-inflammatory and analgesic agents that could improve pain management in horses. The objective of the present study was to evaluate the anti-nociceptive effect of a single-dose intravenous administration of the sEH inhibitor trans-4-{4-[3-(4-trifluro-methoxy-phenyl)-ureido]-cyclohexyloxy}-benzoic acid (t-TUCB) using an adjustable heart bar shoe (a-HBS) model of lameness. We hypothesized that t-TUCB would improve objective and subjective lameness measures compared to the control. MATERIALS AND METHODS: Reversible lameness was induced in 8 horses for 24 h using an a-HBS in a randomized, crossover design. A vehicle-control placebo (DMSO) or t-TUCB (1 mg/kg) was intravenously administered at time 0 following a baseline induced lameness evaluation. Blood was collected and lameness was objectively measured with an inertial sensor system at 0-, 1-, 3-, 6-, 12-, and 24-h time points. Front-facing videos were obtained at each time point for subjective evaluation by three blinded evaluators using a visual analog scale (VAS). RESULTS: Treatment with t-TUCB significantly decreased (i.e. improved) lameness compared to placebo at 1-h and compared to baseline at 1-, 3-, and 6-h following administration. Lameness significantly increased (i.e. worsened) from baseline in placebo-treated horses 12 h after administration. All horses returned to baseline soundness within 24 h of reversing lameness. CONCLUSION: Treatment with single-dose IV administration of t-TUCB improved lameness induced by the a-HSB, suggesting that t-TUCB has anti-nociceptive effects in horses. CLINICAL RELEVANCE: The soluble epoxide hydrolase inhibitor, t-TUCB, is a promising novel analgesic for horses.

Comparative effects of nonsteroidal anti-inflammatory drugs at castration and tail-docking in neonatal piglets.

This study assessed the efficacy of meloxicam, flunixin, and ketoprofen in piglets undergoing routine castration and tail-docking. Six-day-old male piglets (8/group) received one of five randomized treatments: intramuscular saline (SAL PROC), meloxicam (MEL; 0.4 mg/kg), flunixin (FLU; 2.2 mg/kg), ketoprofen (KETO; 3.0 mg/kg) or sham (SAL SHAM; saline injection, no processing). Two hours post-dose, piglets were castrated and tail-docked. Plasma cortisol, interstitial fluid (ISF) prostaglandin E2 (PGE2) and activity levels via Actical® monitoring were used to estimate pain. SAL SHAM and FLU exhibited lower cortisol concentrations than SAL PROC at the time of processing (p = 0.003 and p = 0.049, respectively), and all NSAIDs exhibited lower PGE2 than SAL PROC at 3.69 hours (MEL p = 0.050; FLU p = 0.043 and KETO p = 0.031). While not statistically significant, PGE2 was higher in SAL PROC piglets vs. other treatment groups at most time points. There was also a high degree of variability between piglets, especially for SAL PROC. Activity levels were significantly decreased at multiple time points in SAL PROC and MEL piglets following processing. However, FLU and KETO piglets had increased activity levels closer to that of the SAL SHAM group, suggesting that these NSAIDs are more effective than MEL in providing analgesia. These results demonstrate that management strategies including administration of intramuscular flunixin or ketoprofen to reduce pain associated with processing will likely improve piglet health and welfare in the United States.

Larazotide acetate induces recovery of ischemia-injured porcine jejunum via repair of tight junctions.

Intestinal ischemia results in mucosal injury, including paracellular barrier loss due to disruption of tight junctions. Larazotide acetate (LA), a small peptide studied in Phase III clinical trials for treatment of celiac disease, regulates tight junctions (TJs). We hypothesized that LA would dose-dependently hasten recovery of intestinal ischemic injury via modulation of TJs. Ischemia-injured tissue from 6-8-week-old pigs was recovered in Ussing chambers for 240-minutes in the presence of LA. LA (1 µM but not 0.1 µM or 10 µM) significantly enhanced transepithelial electrical resistance (TER) above ischemic injured controls and significantly reduced serosal-to-mucosal flux LPS (P<0.05). LA (1 µM) enhanced localization of the sealing tight junction protein claudin-4 in repairing epithelium. To assess for the possibility of fragmentation of LA, an in vitro enzyme degradation assay using the brush border enzyme aminopeptidase M, revealed generation of peptide fragments. Western blot analysis of total protein isolated from uninjured and ischemia-injured porcine intestine showed aminopeptidase M enzyme presence in both tissue types, and mass spectrometry analysis of samples collected during ex vivo analysis confirmed formation of LA fragments. Treatment of tissues with LA fragments had no effect alone, but treatment with a fragment missing both amino-terminus glycines inhibited barrier recovery stimulated by 1 µM LA. To reduce potential LA inhibition by fragments, a D-amino acid analog of larazotide Analog #6, resulted in a significant recovery response at a 10-fold lower dose (0.1 µM) similar in magnitude to that of 1 µM LA. We conclude that LA stimulates repair of ischemic-injured epithelium at the level of the tight junctions, at an optimal dose of 1 µM LA. Higher doses were less effective because of inhibition by LA fragments, which could be subverted by chirally-modifying the molecule, or microdosing LA.

Longitudinal impact of a pre-populated default quantity on emergency department opioid prescriptions.

OBJECTIVE: Previously published studies indicate that a pre-populated default quantity may decrease opioid amounts on discharge prescriptions from the emergency department (ED). However, the longitudinal effect of defaulted quantities has not been described in the literature. METHODS: A retrospective review of electronic health record data from visits to 4 hospital EDs in a community health system examined opioid prescription dispense quantities 3.5 years pre- and 6.5 years post-implementation of a defaulted dispense quantity of seventeen. The primary purpose was to determine the percentage of ED discharge opioid prescriptions containing the prepopulated default dispense quantity after implementation. The longitudinal effect of a default quantity implementation on the average quantity prescribed (normalized per 1000 visits) was examined by comparing the pre-implementation period (January 1, 2009-July 31, 2012) to the post-implementation period (August 1, 2012-June 30, 2018). RESULTS: After implementation in 2012, the acceptance rate of the default dispense quantity increased each year, up to 48% in 2016 and maintained through 2018. A significant decrease in prescribed opioid quantities post-default quantity implementation was sustained, with the average quantity prescribed from 2015-2018 maintained at 17 or lower. CONCLUSION: A pre-populated default quantity impacts discharge opioid prescribing as evidenced by a high sustained rate of prescriber utilization over years and reduction in the per prescription average pill quantity. The acceptance of a pre-populated default quantity may allow for selection of even a lower quantity to influence prescribing patterns of opioid analgesics.

Evaluation of air-free glassware using the ketyl test.

Air-free glassware and techniques are used in most organometallic research laboratories, but their relative effectiveness has not been well studied. We report a method for quantitatively comparing air and moisture exclusion by air-free glassware. Samples of benzophenone dianion in various glassware are monitored by video as their purple color is quenched. Numerical values for time until colorless are extracted from the video data and interpreted as a relative measure of air and moisture exclusion. Low pressure/vacuum (LPV) NMR tubes demonstrated by far the best performance, compared to a variety of Schlenk flasks and standard NMR tubes. The findings are of immediate practical use for synthetic chemists, and the evaluation methods can be used by individual laboratories to assess other conditions for air and moisture exclusion.

Sterility and concentration of liposomal bupivacaine single-use vial when used in a multiple-dose manner.

OBJECTIVE: To evaluate the sterility of bupivacaine liposome injectable suspension (Nocita®) used in a multiple-dose fashion for 5 days. STUDY DESIGN: Triplicate liposomal bupivacaine vials were stored under two conditions, (1) room temperature (24°C) and (2) refrigerated temperature (5°C). A 3-mL aliquot was withdrawn from each vial daily. Samples were inoculated in tryptic soy broth in triplicate and then incubated for 24 hours at 37°C and subcultured every 48 hours onto blood agar and Sabouraud dextrose agar, respectively. Separate 1.5-mL aliquots of liposomal bupivacaine were centrifuged at 3500 g to separate liposome-encapsulated bupivacaine from the solution. Concentration of unencapsulated bupivacaine was analyzed via high-pressure liquid chromatography. Data were analyzed by using mixed effects procedure with multiple comparisons. SAMPLE POPULATION: Ten 20-mL vials of bupivacaine liposome injectable suspension stored under two conditions, (1) room temperature (24°C) and (2) refrigerated temperature (5°C). RESULTS: Five days of repeated withdrawal from the single-use vials yielded no bacterial growth. One control vial, which was opened and punctured once on the last day of the experiment, yielded fungal growth of an Aspergillus spp, likely an environmental contaminant. The concentration of free bupivacaine did not significantly differ until the fifth day of sampling. CONCLUSION: When aseptic technique was used, liposomal bupivacaine remained sterile for 5 days. Concentrations of free bupivacaine were unchanged from baseline for 4 days in both refrigerated and room temperature conditions. CLINICAL SIGNIFICANCE: Single-use liposomal bupivacaine vials can be used extralabel in a multiple-dose fashion for up to 4 days when stored either refrigerated or room temperature when sterile technique is used.

Nutrient Regulation by Continuous Feeding for Large-scale Expansion of Mammalian Cells in Spheroids.

In this demonstration, spheroids formed from the ß-TC6 insulinoma cell line were cultured as a model of manufacturing a mammalian islet cell product to demonstrate how regulating nutrient levels can improve cell yields. In previous studies, bioreactors facilitated increased culture volumes over static cultures, but no increase in cell yields were observed. Limitations in key nutrients such as glucose, which were consumed between batch feedings, can lead to limitations in cell expansion. Large fluctuations in glucose levels were observed, despite the increase in glucose concentrations in the media. The use of continuous feeding systems eliminated fluctuations in glucose levels, and improved cell growth rates when compared with batch fed static and SSB culture methods. Additional increases in growth rates were observed by adjusting the feed rate based on calculated nutrient consumption, which allowed the maintenance of physiological glucose over three weeks in culture. This method can also be adapted for other cell types.

Nutrient regulation by continuous feeding removes limitations on cell yield in the large-scale expansion of Mammalian cell spheroids.

Cellular therapies are emerging as a standard approach for the treatment of several diseases. However, realizing the promise of cellular therapies across the full range of treatable disorders will require large-scale, controlled, reproducible culture methods. Bioreactor systems offer the scale-up and monitoring needed, but standard stirred bioreactor cultures do not allow for the real-time regulation of key nutrients in the medium. In this study, ß-TC6 insulinoma cells were aggregated and cultured for 3 weeks as a model of manufacturing a mammalian cell product. Cell expansion rates and medium nutrient levels were compared in static, stirred suspension bioreactors (SSB), and continuously fed (CF) SSB. While SSB cultures facilitated increased culture volumes, no increase in cell yields were observed, partly due to limitations in key nutrients, which were consumed by the cultures between feedings, such as glucose. Even when glucose levels were increased to prevent depletion between feedings, dramatic fluctuations in glucose levels were observed. Continuous feeding eliminated fluctuations and improved cell expansion when compared with both static and SSB culture methods. Further improvements in growth rates were observed after adjusting the feed rate based on calculated nutrient depletion, which maintained physiological glucose levels for the duration of the expansion. Adjusting the feed rate in a continuous medium replacement system can maintain the consistent nutrient levels required for the large-scale application of many cell products. Continuously fed bioreactor systems combined with nutrient regulation can be used to improve the yield and reproducibility of mammalian cells for biological products and cellular therapies and will facilitate the translation of cell culture from the research lab to clinical applications.