HBD-1 production by patient unburned skin tissues in cultures supplemented with CD31⁺ IMC culture fluids.

<p>Patient #16∼#18 CD31⁺ IMC (1×10⁶ cells/ml) were individually treated with or without glycyrrhizin (100 µg/ml) for 12 hours. After washing with media, these cells were cultured for an additional 36 hours. Culture fluids harvested from the cultures supplemented with or without glycyrrhizin were added (5∼20%, v/v) to cultures of patient unburned skin tissues (0.2×0.2 cm), and cultured for 48 hours. Culture fluids harvested were assayed for HBD-1 by ELISA. *<i>P</i><0.05; **<i>P</i><0.01 vs. cultures without glycyrrhizin.</p

Culture fluids of lineage⁻CD34⁺CD31⁺ cells are inhibitory on HBD-1 production by NHEK. A.

<p>Inhibition of HBD production in NHEK cultures supplemented with culture fluids of lineage⁻CD34⁺CD31⁺ cells. Culture fluids were harvested 48 hours after cultivation of lineage⁻CD34⁺CD31⁺ cells (1×10⁶ cells/ml) derived from burn patients #8, #9, #13, #14 and #15 (open circles). The culture fluids of lineage⁻CD34⁺CD31⁻ cells of healthy donors #1∼#5 (filled circles) were utilized as a control. Five to 40% (v/v) of these culture fluids were added to cultures of NHEK (1×10⁵ cells/ml). Culture fluids harvested from NHEK cultures were assayed for HBD-1 by ELISA. * <i>P</i><0.01 vs control. <b>B–D.</b> Production of IL-10, IL-13 and CCL2 by lineage⁻CD34⁺CD31⁺ cells. Lineage⁻CD34⁺CD31⁺ cells (1×10⁵ cells/ml, open circles) isolated from peripheral blood of burn patients #8, #9, #13, #14 and #15 were cultured for 12 to 48 hours. As controls, peripheral blood lineage⁻CD34⁺CD31⁻ cells of healthy donors #1∼#5 (filled circles) were cultured under the same conditions. Supernatants obtained were assayed for IL-10 (<b>B</b>), IL-13 (<b>C</b>) and CCL2 (<b>D</b>) by ELISA. * <i>P</i><0.05, ** <i>P</i><0.01 vs lineage⁻CD34⁺CD31⁻ cells.</p

HBD-1 production and mRNA expression by NHEK cultured with peripheral blood lineage⁻CD34⁺ cells that were isolated from both severely burned patients and healthy donors. A.

<p>HBD-1 production. Lineage⁻CD34⁺ cells (1×10⁵ cells/ml, upper chamber), isolated from 5 healthy donors (#1∼#5) and 20 burn patients (#1∼#20), were transwell-cultured with NHEK (1×10⁵ cells/ml, lower chamber) for 36 hours. After removal of the upper chamber, NHEK in the lower chamber were cultured for an additional 36 hours. Culture fluids obtained were assayed for HBD-1 by ELISA. <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0082926#pone-0082926-g001" target="_blank">Fig. 1A-1</a> shows independent experiments performed using blood specimens from 5 healthy donors and 20 burn patients, and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0082926#pone-0082926-g001" target="_blank">Fig. 1A-2</a> shows mean ± SEM of the results shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0082926#pone-0082926-g001" target="_blank">Fig. 1A-1</a>. * <i>P</i><0.001 vs control. <b>B and C.</b> HBD-1 mRNA expression. Lineage⁻CD34⁺ cells (1×10⁵ cells/ml, upper chamber), isolated from peripheral blood of 5 healthy donors (#1∼#5, <b>B</b>) and 5 severely burn patients (#1, #16, #17, #18, #19, <b>C</b>), were transwell-cultured with NHEK (1×10⁵ cells/ml, lower chamber) for 36 hours. After removal of the upper chamber, NHEK in the lower chamber were analyzed for HBD-1 mRNA by RT-PCR.</p

Additional file 1 of An assessment of physician assistant student diversity in the United States: a snapshot for the healthcare workforce

Additional file 1: Supplemental Appendix 1. Top PA Performers Ranked by Number of Graduates from 2014-2018

Additional file 2 of An assessment of physician assistant student diversity in the United States: a snapshot for the healthcare workforce

Additional file 2: Supplemental Appendix 2. Top PA Performers Ranked by Proportion of Graduates from 2014-2018

Structure‑H Methane + 1,1,2,2,3,3,4-Heptafluorocyclopentane Mixed Hydrate at Pressures up to 373 MPa

Thermodynamic stability boundary of structure-H hydrates with large guest species and methane (CH₄) at extremely high pressures has been almost unclear. In the present study, the four-phase equilibrium relations in the structure-H CH₄ + 1,1,2,2,3,3,4-heptafluorocyclopentane (1,1,2,2,3,3,4-HFCP) mixed hydrate system were investigated in a temperature range of (281.05 to 330.12) K and a pressure range up to 373 MPa. The difference between equilibrium pressures in the structure-H CH₄ + 1,1,2,2,3,3,4-HFCP mixed hydrate system and the structure-I simple CH₄ hydrate system gets larger with increase in temperature. The structure-H CH₄ + 1,1,2,2,3,3,4-HFCP mixed hydrate survives even at 330 K and 373 MPa without any structural phase transition. The maximum temperature where the structure-H CH₄ + 1,1,2,2,3,3,4-HFCP mixed hydrate is thermodynamically stable is likely to be beyond that of the structure-H simple CH₄ hydrate

Associations Between Drug Use and Motorcycle Helmet Use in Fatal Crashes

<div><p><b>Objective:</b> Helmet use reduces mortality risk for motorcyclists, regardless of drug and alcohol use. However, the association between drug use and motorcycle helmet utilization is not well known. This study examines the relationship between drug use and motorcycle helmet use among fatally injured motorcycle riders.</p><p><b>Methods:</b> Using data from the 2005–2009 Fatality Analysis Reporting System (FARS), we examined the association between drug use and motorcycle helmet use in a multivariable logistic regression analysis of 9861 fatally injured motorcycle riders in the United States.</p><p><b>Results:</b> For fatally injured motorcycle riders, use of alcohol, marijuana, or other drugs was associated with increased odds of not wearing a motorcycle helmet, controlling for the effects of state motorcycle helmet laws and other confounding variables. Predicted probabilities indicate that helmet use substantially decreases among fatally injured riders mixing alcohol with marijuana and other drugs. Furthermore, the likelihood of helmet use between marijuana-only users and other drug users is virtually the same across all blood alcohol content (BAC) levels.</p><p><b>Conclusions:</b> This study provides evidence that alcohol, marijuana, and other drug use is associated with not wearing a motorcycle helmet in fatal motorcycle crashes. There is a clear need for additional prevention and intervention efforts that seek to change helmet and drug use norms among motorcycle riders.</p></div

CCL2 and IL-10 produced by burn patient lineage⁻CD34⁺ CD31⁺ cells are responsible for their suppressor cell activities on HBD-1 production by NHEK. A.

<p>Effect of mAbs against IL-10, CCL2 and IL-13 on the suppressor activities of lineage⁻CD34⁺CD31⁺ cell-culture fluids on HBD production by NHEK. NHEK (1×10⁵ cells/ml) were cultured with fresh media supplemented with 20% (v/v) of the culture fluids that were previously treated with mAb for IL-10, IL-13 and CCL2 (2.5 µg/ml each) or a combination of these mAbs. Forty-eight hours after cultivation, culture fluids harvested were assayed for HBD-1. <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0082926#pone-0082926-g004" target="_blank">Figure 4A</a> displays one of the representative results shown in independent experiments using culture fluids of 5 different lineage⁻CD34⁺CD31⁺ cell preparations derived from burn patients #8, #9, #13, #14 and #15. <b>B.</b> Effect of rIL-10 and rCCL2 on HBD-1 production by NHEK. NHEK (1×10⁵ cells/ml) were treated with rIL-10 (1 ng/ml), rCCL2 (10 ng/ml) or a combination of rIL-10 and rCCL2. Culture fluids obtained 24 hours after treatment were assayed for HBD-1. * <i>P</i><0.01, ** <i>P</i><0.001 vs NHEK cultured with media.</p

HBD-1 production in patient chimeras treated with glycyrrhizin.

<p><b>A.</b> Patient chimeras created with burn patient #1∼#3 unburned skin tissues and their CD31⁺ IMC were treated i.p. with glycyrrhizin (10 mg/kg) 6 and 24 hours after the IMC inoculation. As a control, the chimeras were treated with saline (0.2 ml/mouse). Twenty-four hours after the final glycyrrhizin treatment, 5 skin biopsies were obtained from grafted site tissues, homogenized together, and assayed for HBD-1 by ELISA. A white bar shows the results obtained from a group of mice grafted with skin alone. Fig. 2A-1 shows 3 independent experiments performed using skin and blood specimens from 3 patients, and Fig. 2A-2 shows mean ± SEM of the results shown in Fig. 2A-1. **<i>P</i><0.01 vs saline-treated control. <b>B.</b> The recovery of HBD-1 production in patient chimeras treated with glycyrrhizin. The chimeras created with unburned skin tissues from patients #4∼#7 and their CD31⁺ IMC were treated twice with 1 to 10 mg/kg of glycyrrhizin. As controls, the chimeras were treated with saline (0.2 ml/mouse). Twenty-four hours after the treatment, 5 skin biopsies were obtained from grafted site tissues, homogenized together, and assayed for HBD-1 by ELISA. A white bar shows the results obtained from a group of mice grafted with skin alone (mean ± SEM of the 4 independent experiments). *<i>P</i><0.05; **<i>P</i><0.01 vs saline-treated control.</p

Effect of glycyrrhizin on the resistance of chimeras against sepsis stemming from <i>P. aeruginosa</i> i.d. infections.

<p><b>A.</b> Survival of patient chimera substitutes i.d. infected with 20 LD₅₀ of <i>P. aeruginosa</i>. The chimeras exposed to the pathogen were treated i.p. with 10 mg/kg (open circles) and 3 mg/kg (open squires) of glycyrrhizin or saline (0.2 ml/mouse, control, open triangles). Each infection experiment was performed by a group of 2–3 mice, and it was repeated 3 times. The results obtained were combined and displayed in the figure. <b>B.</b> Growth of pathogen in kidneys of patient chimeras i.d. infected with 20 LD₅₀ of <i>P. aeruginosa</i>. Patient chimeras created with unburned skin tissues from patients #8∼#10 and their CD31⁺ IMC were exposed to the pathogen, and treated with glycyrrhizin (10 mg/kg) or saline (0.2 ml/mouse, control). Two days after infection, the growth of <i>P. aeruginosa</i> in kidneys of these chimeras was measured by a standard colony counting method. A white bar shows the results obtained from a group of mice grafted with skin alone. Fig. 3B-1 shows 3 independent experiments performed using skin and blood specimens from 3 patients, and Fig. 3B-2 shows mean ± SEM of the results shown in Fig. 3B-1. **<i>P</i><0.01 vs saline-treated control.</p