Determining If Sex Bias Exists in Human Surgical Clinical Research

Sex is a variable that is poorly controlled for in clinical research

Use of complete medication history to identify and correct transitions-of-care medication errors at psychiatric hospital admission.

Methods for categorizing the scale and severity of medication errors corrected by pharmacy staff during admission medication reconciliation using complete medication history continue to evolve. We established a rating scale that is effective for generating error reports to health system quality leadership. These reports are needed to quantify the value of investment in transitions-of-care pharmacy staff. All medication errors that were reported by pharmacy staff in the admission medication reconciliation process during a period of 6 months were eligible for inclusion. Complete medication history data source was utilized by admitting providers and all pharmacist staff and a novel medication error scoring methodology was developed. This methodology included: medication error category, medication error type, potential medication error severity, and medication non-adherence. We determined that 82 medication errors were detected from 72 patients and assessed that 74 of these errors may have harmed patients if they were not corrected through pharmacist intervention. Most of these errors were dosage discrepancies and omissions. With hospital system budgets continually becoming leaner, it is important to measure the effectiveness and value of staff resources to optimize patient care. Pharmacists performing admission medication reconciliation can detect subtle medication discrepancies that may be overlooked by other clinician types. This methodology can serve as a foundation for error reporting and predicting the severity of adverse drug events

Determining If Sex Bias Exists in Human Surgical Clinical Research

Sex is a variable that is poorly controlled for in clinical research

Remodeling of Tight Junctions and Enhancement of Barrier Integrity of the CACO-2 Intestinal Epithelial Cell Layer by Micronutrients.

The micronutrients zinc, quercetin, butyrate, indole and berberine were evaluated for their ability to induce remodeling of epithelial tight junctions (TJs) and enhance barrier integrity in the CACO-2 gastrointestinal epithelial cell culture model. All five of these chemically very diverse micronutrients increased transepithelial electrical resistance (Rt) significantly, but only berberine also improved barrier integrity to the non-electrolyte D-mannitol. Increases of Rt as much as 200% of untreated controls were observed. Each of the five micronutrients also induced unique, signature-like changes in TJ protein composition, suggesting multiple pathways (and TJ arrangements) by which TJ barrier function can be enhanced. Decreases in abundance by as much as 90% were observed for claudin-2, and increases of over 300% could be seen for claudins -5 and -7. The exact effects of the micronutrients on barrier integrity and TJ protein composition were found to be highly dependent on the degree of differentiation of the cell layer at the time it was exposed to the micronutrient. The substratum to which the epithelial layer adheres was also found to regulate the response of the cell layer to the micronutrient. The implications of these findings for therapeutically decreasing morbidity in Inflammatory Bowel Disease are discussed

The effect of six micronutrients on CACO-2 transepithelial electrical resistance.

<p>1A: 7-Day post-confluent CACO-2 cell layers on Millipore PCF filters were refed in control medium (apical and basal-lateral compartments) or medium containing 50μM, 100μM, or 200μM berberine chloride 12–17 hrs prior to electrical measurements. Data shown represents the mean ± standard error of 9 cell layers per condition (3 experiments, 3 cell layers per experiment). 1B: 7-Day post-confluent CACO-2 cell layers on Millipore PCF filters were refed, as above, in control medium or medium containing 1μM, 10μM, or 100μM Nicotine 48 hrs prior to electrical measurements. Data shown represents the mean ± standard error of 6 cell layers per condition (2 experiments, 3 cell layers per experiment). 1C: 4-Day post-confluent CACO-2 cell layers on Millipore PCF filters were refed, as in A, in control medium or medium containing 0.5mM, 2.0mM, or 5mM sodium butyrate 72 hrs prior to electrical measurements. Data shown represents the mean ± standard error of 6 cell layers per condition (2 experiments, 3 cell layers per experiment). 1D: 7-Day post-confluent CACO-2 cell layers on Millipore PCF filters were refed, as in A, in control medium or medium containing 100μM, 200μM, or 400μM quercetin 48hrs prior to electrical measurements. Data shown represents the mean ± standard error of 6 cell layers per condition (2 experiments, 3 cell layers per experiment). 1E: 7-Day post-confluent CACO-2 cell layers on Millipore PCF filters were refed, as in A, in control medium or medium containing 0.5mM, 1.0mM, or 2.0mM indole 48hrs prior to electrical measurements. Data shown represents the mean ± standard error of 9 cell layers per condition (3 experiments, 3 cell layers per experiment). 1F: 7-Day post-confluent CACO-2 cell layers on Millipore PCF filters were refed, as in A, in control medium or medium containing 50μM, 100μM, or 150μM zinc sulfate 48hrs prior to electrical measurements. Data shown represents the mean ± standard error of 12 cell layers for both the control and 100μM conditions, and 4 cell layers for both the 50μM and 150μM conditions. In all cases, data represents the percent of control resistance normalized for each experiment. * indicates P < 0.05; ** indicates P < 0.01; *** indicates P < 0.001 (one-way ANOVA followed by Dunnett’s post hoc testing versus control).</p

Effect of the Filter Substratum on CACO-2 Transepithelial Barrier Function Properties.

<p>CACO-2 cell layers were seeded at identical densities on Millicell PCF and Millicell HA units as described in Materials and Methods. After 7 days, cell layers were refed in apical and basal-lateral compartments with control medium prior to electrical measurements and ¹⁴C-D-mannitol flux studies. Data shown is expressed as the mean ± standard error of 8 cell layers per condition (2 experiments, 4 cell layers per condition).</p><p>NS indicates non significance.</p><p>*** indicates P<0.001 (Student’s t test, two-tailed).</p><p>Effect of the Filter Substratum on CACO-2 Transepithelial Barrier Function Properties.</p

Effect of zinc on CACO-2 tight junctional proteins as a function of the differentiation state of the CACO-2 cell layer.

<p>3-day, 7-day, and 21-day post-confluent CACO-2 cell layers in Falcon 75 cm² culture flasks were refed with control medium or medium containing 100μM Zinc 48 hrs before harvesting in lysis buffer. Further steps were performed as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0133926#pone.0133926.t001" target="_blank">Table 1</a>. Data represents the percent of the zinc condition relative to the normalized control for each age cell layer. Data shown is expressed as the mean ± standard error for an n = 4 cell layers in all cases. NS indicates non significance. * indicates P < 0.05; ** indicates P < 0.01; *** indicates P < 0.001 (Student’s t test, two-tailed).</p

Effects of Individual Micronutrients on CACO-2 Tight Junctional Proteins.

<p>Summary of effects of various micronutrients on a panel of eight tight junctional proteins in CACO-2 cell layers. Post-confluent CACO-2 cell layers in Falcon 75 cm² culture flasks were refed on both sides with control medium or medium containing 100μM Zinc 48 hrs before harvesting in lysis buffer. These total cell lysates were analyzed by PAGE followed by immunoblotting as described in Methods. Immunoblots were probed with primary antisera against specific tight junctional antigens also as described. Densitometry was performed on developed blots to quantitate band densities. Data shown represents the mean ± standard error for an n = 3 cell layers in all cases. P values are listed for instances of statistical significance (Student’s t test [two tailed] to at least the P<0.05 level). NS indicates non significance. This same procedure was used for quercetin (400μM), butyrate (5.0mM), berberine (100μM), and indole (1.0mM) at the concentrations that provided maximal barrier enhancement and for the same time periods used in Figs <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0133926#pone.0133926.g001" target="_blank">1</a> and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0133926#pone.0133926.g002" target="_blank">2</a>. Significant increases are highlighted in green while significant decreases are highlighted in yellow to emphasize the distinct overall pattern specific to each micronutrient.</p><p>Effects of Individual Micronutrients on CACO-2 Tight Junctional Proteins.</p

The effect of six micronutrients on CACO-2 transepithelial flux of ¹⁴C-D-Mannitol.

<p>In all cases, after electrical measurements, the same CACO-2 cell layers represented in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0133926#pone.0133926.g001" target="_blank">Fig 1</a> were used to perform radiotracer flux studies with 0.1mM, 0.20 μCi/ml ¹⁴C-D-mannitol, as described in Materials and Methods. Data represents the percent of control flux rate normalized for each experiment, and is expressed as the mean ± standard error for the total number of cell layers per condition, as detailed in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0133926#pone.0133926.g001" target="_blank">Fig 1</a>. * indicates P < 0.05, ** indicates P < 0.01 and *** indicates P < 0.001 (one-way ANOVA followed by Dunnett’s post hoc testing versus control).</p

Differential effect of quercetin treatment on the tight junctional proteins of 7-day vs. 1-day post-confluent CACO-2 cell layers.

<p>1-day and 7-day post-confluent CACO-2 cell layers in Falcon 75 cm² culture flasks were refed with control medium or medium containing 400μM quercetin 48 hrs before harvesting in lysis buffer. Further steps were performed as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0133926#pone.0133926.t001" target="_blank">Table 1</a>. Data represents the percentage of band density of the no-quercetin control for that CACO-2 cell layer. Data shown is expressed as the mean ± standard error for an n = 3 cell layers in all cases. * indicates P < 0.05 for 1-day vs. 7-day changes in TJ protein (Student’s t test, two-tailed.</p