On the development and validation of large language model-based classifiers for identifying social determinants of health.

The assessment of social determinants of health (SDoH) within healthcare systems is crucial for comprehensive patient care and addressing health disparities. Current challenges arise from the limited inclusion of structured SDoH information within electronic health record (EHR) systems, often due to the lack of standardized diagnosis codes. This study delves into the transformative potential of large language models (LLM) to overcome these challenges. LLM-based classifiers-using Bidirectional Encoder Representations from Transformers (BERT) and A Robustly Optimized BERT Pretraining Approach (RoBERTa)-were developed for SDoH concepts, including homelessness, food insecurity, and domestic violence, using synthetic training datasets generated by generative pre-trained transformers combined with authentic clinical notes. Models were then validated on separate datasets: Medical Information Mart for Intensive Care-III and our institutional EHR data. When training the model with a combination of synthetic and authentic notes, validation on our institutional dataset yielded an area under the receiver operating characteristics curve of 0.78 for detecting homelessness, 0.72 for detecting food insecurity, and 0.83 for detecting domestic violence. This study underscores the potential of LLMs in extracting SDoH information from clinical text. Automated detection of SDoH may be instrumental for healthcare providers in identifying at-risk patients, guiding targeted interventions, and contributing to population health initiatives aimed at mitigating disparities.

A Point-Based Risk Calculator for Mortality After Hepatectomy.

BACKGROUND: Preoperative risk stratification for hepatectomy patients can aid clinical decision making. The objective of this retrospective cohort study was to determine postoperative mortality risk factors and develop a score-based risk calculator using a limited number of preoperative predictors to estimate mortality risk in patients undergoing hepatectomy. METHODS: Data were collected from patients that underwent hepatectomy from the National Surgical Quality Improvement Program dataset from 2014 to 2020. Baseline characteristics were compared between survival and 30-day mortality cohorts using the χ 2 test. Next, the data were split into a training set to build the model and a test set to validate the model. A multivariable logistic regression model modeling 30-day postoperative mortality was trained on the training set using all available features. Next, a risk calculator using preoperative features was developed for 30-day mortality. The results of this model were converted into a score-based risk calculator. A point-based risk calculator was developed that predicted 30-day postoperative mortality in patients who underwent hepatectomy surgery. RESULTS: The final dataset included 38,561 patients who underwent hepatectomy. The data were then split into a training set from 2014 to 2018 (n = 26,397) and test set from 2019 to 2020 (n = 12,164). Nine independent variables associated with postoperative mortality were identified and included age, diabetes, sex, sodium, albumin, bilirubin, serum glutamic-oxaloacetic transaminase (SGOT), international normalized ratio, and American Society of Anesthesiologists classification score. Each of these features were then assigned points for a risk calculator based on their odds ratio. A univariate logistic regression model using total points as independent variables were trained on the training set and then validated on the test set. The area under the receiver operating characteristics curve on the test set was 0.719 (95% confidence interval, 0.681-0.757). CONCLUSIONS: Development of risk calculators may potentially allow surgical and anesthesia providers to provide a more transparent plan to support patients planned for hepatectomy.

Racial and Ethnic Differences in the Receipt of Regional Anesthesia Among Patients Undergoing Thoracic Surgery.

OBJECTIVES: The objective of this study was to assess differences in the use of perioperative regional anesthesia for thoracic surgery based on race and ethnicity. DESIGN: This retrospective cohort study used data from the American College of Surgeons National Surgical Quality Improvement Program from 2015 to 2020. The study authors applied a multivariate logistic regression in which the dependent variable was the primary endpoint (regional versus no regional anesthesia). The primary independent variables were race and ethnicity. SETTING: Multiple healthcare systems in the United States. PARTICIPANTS: Participants were ≥18 years of age and undergoing thoracic surgery. INTERVENTIONS: Regional anesthesia. MEASUREMENTS AND MAIN RESULTS: On adjusted multivariate analysis, Hispanic patients had lower odds (odds ratio [OR] 0.61, 95% CI 0.46-0.80, p = 0.0003) of receiving regional anesthesia for postoperative pain control compared to non-Hispanic patients. There was no significant difference in the odds of regional anesthesia when comparing racial cohorts (ie, White, Black, Asian, or other). CONCLUSIONS: There were differences observed in the provision of regional anesthesia for thoracic surgery among ethnic groups. Although the results of this study should not be taken as evidence for healthcare disparities, it could be used to support hypotheses for future studies that aim to investigate causes of disparities and corresponding patient outcomes.

Racial and Ethnic Differences in the Use of Regional Anesthesia for Patients Undergoing Total Knee Arthroplasty.

BACKGROUND: There is an increasing body of evidence that suggests racial and ethnic disparities exist in medical care. In the field of anesthesiology, few studies have investigated the association of race and ethnicity with the provision of regional anesthesia for patients undergoing total knee arthroplasty. This analysis queried a large national surgical database to determine whether there were racial or ethnic differences in the administration of peripheral nerve blocks for patients undergoing total knee arthroplasty. METHODS: In this retrospective cohort study, data were collected from a large national database during the years 2017-2019. Multivariable logistic regressions were used to measure the association of race and ethnicity with utilization of regional anesthesia. The participants for the study were patients 18 years or older undergoing total knee arthroplasty. RESULTS: Our primary finding was that among patients undergoing total knee arthroplasty, Black patients had lower odds (adjusted odds ratio [aOR]: 0.93, 99% confidence interval [CI]: 0.89-0.98) of receiving regional anesthesia than White patients. Also, Hispanic patients had lower odds (aOR: 0.88, 99% CI: 0.83-0.94) of receiving regional anesthesia than non-Hispanic patients. Native Hawaiian/Pacific Islander patients had increased odds (aOR: 2.04, 99% CI: 1.66-2.51) of receiving regional anesthesia. CONCLUSION: This study demonstrated that there might be racial and ethnic differences in the provision of regional anesthesia for patients undergoing total knee arthroplasty. These differences underscore the need for more studies aimed at equitable access to high quality and culturally competent health care.

A Retrospective Analysis Investigating Whether Case Volume Experience of the Anesthesiologist Correlates with Intraoperative Efficiency for Joint Arthroplasty.

The objective of this retrospective study was to determine if there was an association between anesthesiology experience (e.g. historic case volume) and operating room (OR) efficiency times for lower extremity joint arthroplasty cases. The primary outcome was time from patient in the OR to anesthesia ready (i.e. after spinal or general anesthesia induction was complete). The secondary outcomes included time from anesthesia ready to surgical incision, and time from incision to closing completed. Mixed effects linear regression was performed, in which the random effect was the anesthesiology attending provider. There were 4,575 patients undergoing hip or knee arthroplasty included. There were 82 unique anesthesiology providers, in which the median [quartile] frequency of cases performed was 79 [45, 165]. On multivariable mixed effects linear regression - in which the primary independent variable (anesthesiologist case volume history for joint arthroplasty anesthesia) was log-transformed - the estimate for log-transformed case volume was - 0.91 (95% confidence interval [CI] -1.62, -0.20, P = 0.01). When modeling time from incision to closure complete, the estimate for log-transformed case volume was - 2.07 (95% -3.54, -0.06, P = 0.01). Thus, when comparing anesthesiologists with median case volume (79 cases) versus those with the lowest case volume (10 cases), the predicted difference in times added up to only approximately 6 min. If the purpose of faster anesthesia workflows was to open up more OR time to increase surgical volume in a given day, this study does not support the supposition that anesthesiologists with higher joint arthroplasty case volume would improve throughput.

Machine Learning Prediction Models to Reduce Length of Stay at Ambulatory Surgery Centers Through Case Resequencing.

The post-anesthesia care unit (PACU) length of stay is an important perioperative efficiency metric. The aim of this study was to develop machine learning models to predict ambulatory surgery patients at risk for prolonged PACU length of stay - using only pre-operatively identified factors - and then to simulate the effectiveness in reducing the need for after-hours PACU staffing. Several machine learning classifier models were built to predict prolonged PACU length of stay (defined as PACU stay ≥ 3 hours) on a training set. A case resequencing exercise was then performed on the test set, in which historic cases were re-sequenced based on the predicted risk for prolonged PACU length of stay. The frequency of patients remaining in the PACU after-hours (≥ 7:00 pm) were compared between the simulated operating days versus actual operating room days. There were 10,928 ambulatory surgical patients included in the analysis, of which 580 (5.31%) had a PACU length of stay ≥ 3 hours. XGBoost with SMOTE performed the best (AUC = 0.712). The case resequencing exercise utilizing the XGBoost model resulted in an over three-fold improvement in the number of days in which patients would be in the PACU past 7pm as compared with historic performance (41% versus 12%, P<0.0001). Predictive models using preoperative patient characteristics may allow for optimized case sequencing, which may mitigate the effects of prolonged PACU lengths of stay on after-hours staffing utilization.

Isoflurane Ameliorates Acute Lung Injury by Preserving Epithelial Tight Junction Integrity.

BACKGROUND: Isoflurane may be protective in preclinical models of lung injury, but its use in patients with lung injury remains controversial and the mechanism of its protective effects remains unclear. The authors hypothesized that this protection is mediated at the level of alveolar tight junctions and investigated the possibility in a two-hit model of lung injury that mirrors human acute respiratory distress syndrome. METHODS: Wild-type mice were treated with isoflurane 1 h after exposure to nebulized endotoxin (n = 8) or saline control (n = 9) and then allowed to recover for 24 h before mechanical ventilation (MV; tidal volume, 15 ml/kg, 2 h) producing ventilator-induced lung injury. Mouse lung epithelial cells were similarly treated with isoflurane 1 h after exposure to lipopolysaccharide. Cells were cyclically stretched the following day to mirror the MV protocol used in vivo. RESULTS: Mice treated with isoflurane following exposure to inhaled endotoxin and before MV exhibited significantly less physiologic lung dysfunction. These effects appeared to be mediated by decreased vascular leak, but not altered inflammatory indices. Mouse lung epithelial cells treated with lipopolysaccharide and cyclic stretch and lungs harvested from mice after treatment with lipopolysaccharide and MV had decreased levels of a key tight junction protein (i.e., zona occludens 1) that was rescued by isoflurane treatment. CONCLUSIONS: Isoflurane rescued lung injury induced by a two-hit model of endotoxin exposure followed by MV by maintaining the integrity of the alveolar-capillary barrier possibly by modulating the expression of a key tight junction protein.

Retrospective Analysis of Perioperative Dexmedetomidine Use in Retina Surgeries: Impact on Postanesthesia Care.

BACKGROUND AND OBJECTIVE: Dexmedetomidine (Precedex®) has been linked to depressive hemodynamic effects and increased length of stay in the post-anesthesia care unit (PACU) when used in ambulatory phacoemulsification procedures. We aimed to determine the prevalence and impact of dexmedetomidine use during ambulatory vitreoretinal procedures. PATIENTS AND METHODS: This retrospective cohort study involved 9,666 adult vitrectomies. Cases were divided into groups by anesthesia type: general anesthesia (GA) and monitored anesthesia care (MAC). For each group, various factors were compared between those who did and did not receive dexmedetomidine. Chi-squared and t tests were used for comparisons. RESULTS: Changes in mean arterial pressure in the MAC group were -1.69 ± 0.23 mmHg for no dexmedetomidine patients and -6.31 ± 0.39 mmHg for dexmedetomidine patients (P < 0.01). In the GA group, mean arterial pressure was -6.1 ± 0.35 mmHg for no dexmedetomidine patients and -11.18 ± 0.88 mmHg for dexmedetomidine patients (P < 0.01). PACU Phase II time in the MAC group was 36.93 ± 0.37 minutes and 40.67 ± 0.86 minutes for no dexmedetomidine and dexmedetomidine patients, respectively (P < 0.01). In the GA group, PACU Phase II time was 58.63 ± 0.95 minutes and 65.19 ± 2.38 minutes for no dexmedetomidine and dexmedetomidine patients, respectively (P < 0.01). CONCLUSIONS: Dexmedetomidine use in vitrectomies was associated with significant PACU delays. These delays may stem from adverse hemodynamic effects. [Ophthalmic Surg Lasers Imaging Retina 2024;55:86-91.].

Development and benchmarking of machine learning models to classify patients suitable for outpatient lower extremity joint arthroplasty.

STUDY OBJECTIVE: Performing hip or knee arthroplasty as an outpatient surgery has been shown to be operationally and financially beneficial for selected patients. By applying machine learning models to predict patients suitable for outpatient arthroplasty, health care systems can better utilize resources efficiently. The goal of this study was to develop predictive models for identifying patients likely to be discharged same-day following hip or knee arthroplasty. DESIGN: Model performance was assessed with 10-fold stratified cross-validation, evaluated over baseline determined by the proportion of eligible outpatient arthroplasty over sample size. The models used for classification were logistic regression, support vector classifier, balanced random forest, balanced bagging XGBoost classifier, and balanced bagging LightGBM classifier. SETTING: The patient records were sampled from arthroplasty procedures at a single institution from October 2013 to November 2021. PATIENTS: The electronic intake records of 7322 knee and hip arthroplasty patients were sampled for the dataset. After data processing, 5523 records were kept for model training and validation. INTERVENTIONS: None. MEASUREMENTS: The primary measures for the models were the F1-score, area under the receiver operating characteristic curve (ROCAUC), and area under the precision-recall curve. To measure feature importance, the SHapley Additive exPlanations value (SHAP) were reported from the model with the highest F1-score. RESULTS: The best performing classifier (balanced random forest classifier) achieved an F1-score of 0.347: an improvement of 0.174 over baseline and 0.031 over logistic regression. The ROCAUC for this model was 0.734. Using SHAP, the top determinant features of the model included patient sex, surgical approach, surgery type, and body mass index. CONCLUSIONS: Machine learning models may utilize electronic health records to screen arthroplasty procedures for outpatient eligibility. Tree-based models demonstrated superior performance in this study.

Chronic endotoxin exposure produces airflow obstruction and lung dendritic cell expansion.

Little is known about the mechanisms of persistent airflow obstruction that result from chronic occupational endotoxin exposure. We sought to analyze the inflammatory response underlying persistent airflow obstruction as a result of chronic occupational endotoxin exposure. We developed a murine model of daily inhaled endotoxin for periods of 5 days to 8 weeks. We analyzed physiologic lung dysfunction, lung histology, bronchoalveolar lavage fluid and total lung homogenate inflammatory cell and cytokine profiles, and pulmonary gene expression profiles. We observed an increase in airway hyperresponsiveness as a result of chronic endotoxin exposure. After 8 weeks, the mice exhibited an increase in bronchoalveolar lavage and lung neutrophils that correlated with an increase in proinflammatory cytokines. Detailed analyses of inflammatory cell subsets revealed an expansion of dendritic cells (DCs), and in particular, proinflammatory DCs, with a reduced percentage of macrophages. Gene expression profiling revealed the up-regulation of a panel of genes that was consistent with DC recruitment, and lung histology revealed an accumulation of DCs in inflammatory aggregates around the airways in 8-week-exposed animals. Repeated, low-dose LPS inhalation, which mirrors occupational exposure, resulted in airway hyperresponsiveness, associated with a failure to resolve the proinflammatory response, an inverted macrophage to DC ratio, and a significant rise in the inflammatory DC population. These findings point to a novel underlying mechanism of airflow obstruction as a result of occupational LPS exposure, and suggest molecular and cellular targets for therapeutic development.

Heme oxygenase-1-derived carbon monoxide enhances the host defense response to microbial sepsis in mice.

Sepsis is characterized by a systemic response to severe infection. Although the inflammatory phase of sepsis helps eradicate the infection, it can have detrimental consequences if left unchecked. Therapy directed against inflammatory mediators of sepsis has shown little success and has the potential to impair innate antimicrobial defenses. Heme oxygenase-1 (HO-1) and the product of its enzymatic reaction, CO, have beneficial antiinflammatory properties, but little is known about their effects on microbial sepsis. Here, we have demonstrated that during microbial sepsis, HO-1-derived CO plays an important role in the antimicrobial process without inhibiting the inflammatory response. HO-1-deficient mice suffered exaggerated lethality from polymicrobial sepsis. Targeting HO-1 to SMCs and myofibroblasts of blood vessels and bowel ameliorated sepsis-induced death associated with Enterococcus faecalis, but not Escherichia coli, infection. The increase in HO-1 expression did not suppress circulating inflammatory cells or their accumulation at the site of injury but did enhance bacterial clearance by increasing phagocytosis and the endogenous antimicrobial response. Furthermore, injection of a CO-releasing molecule into WT mice increased phagocytosis and rescued HO-1-deficient mice from sepsis-induced lethality. These data advocate HO-1-derived CO as an important mediator of the host defense response to sepsis and suggest CO administration as a possible treatment for the disease.

Netropsin improves survival from endotoxaemia by disrupting HMGA1 binding to the NOS2 promoter.

The inducible form of nitric oxide synthase (NOS2) plays an important role in sepsis incurred as a result of infection with Gram-negative bacteria that elaborate endotoxin. The HMGA1 (high-mobility group A1) architectural transcription factor facilitates NOS2 induction by binding a specific AT-rich Oct (octamer) sequence in the core NOS2 promoter via AT-hook motifs. The small-molecule MGB (minor-groove binder) netropsin selectively targets AT-rich DNA sequences and can interfere with transcription factor binding. We therefore hypothesized that netropsin would improve survival from murine endotoxaemia by attenuating NOS2 induction through interference with HMGA1 DNA binding to the core NOS2 promoter. Netropsin improved survival from endotoxaemia in wild-type mice, yet not in NOS2-deficient mice, supporting an important role for NOS2 in the beneficial effects of MGB administration. Netropsin significantly attenuated NOS2 promoter activity in macrophage transient transfection studies and the AT-rich HMGA1 DNA-binding site was critical for this effect. EMSAs (electrophoretic mobility-shift assays) demonstrated that netropsin interferes with HMGA1 NOS2 promoter binding and NMR spectroscopy was undertaken to characterize this disruption. Chemical shift perturbation analysis identified that netropsin effectively competes both HMGA1 DNA-binding AT-hooks from the AT-rich NOS2 promoter sequence. Furthermore, NOESY data identified direct molecular interactions between netropsin and A/T base pairs within the NOS2 promoter HMGA1-binding site. Finally, we determined a structure of the netropsin/NOS2 promoter Oct site complex from molecular modelling and dynamics calculations. These findings represent important steps toward refined structure-based ligand design of novel compounds for therapeutic benefit that can selectively target key regulatory regions within genes that are important for the development of critical illness.

High-mobility group box 1 contributes to lethality of endotoxemia in heme oxygenase-1-deficient mice.

High-mobility group box 1 (HMGB1) is a nuclear protein that has been found to be a critical mediator of lethality in endotoxemia and sepsis. During the systemic inflammatory response, circulating levels of HMGB1 are increased, but in a delayed fashion compared with early inflammatory mediators. To counteract the inflammatory response of endotoxemia, a secondary anti-inflammatory response ensues in an attempt to prevent inflammation-induced tissue injury. One such cytoprotective gene that is induced during endotoxemia is heme oxygenase (HO)-1. HO-1, and its products of heme metabolism, possess anti-inflammatory and antioxidant properties to counter the damaging effects of endotoxemia. In the present study, we wanted to determine whether tissue and circulating levels of HMGB1 are increased further in the absence of HO-1 during endotoxemia, and whether this increase may contribute to the pathobiology of endotoxemia. Lung inflammation, HMGB1 protein levels, and expression of HMGB1 in inflammatory cells were increased in HO-1(-/-) mice compared with HO-1+/+ mice. After the administration of LPS, tissue levels of HMGB1 were not increased further in HO-1(-/-) mice; however, circulating levels of HMGB1 were higher when compared with HO-1+/+ mice. HO-1(-/-) mice treated with a carbon monoxide-releasing molecule or biliverdin showed a reduction in plasma HMGB1, which was associated with a marked improvement in survival. HO-1(-/-) mice given HMGB1-neutralizing antibody showed improvement in survival compared with control antibody. These data suggest that exaggerated circulating levels of HMGB1 contribute to endotoxin-induced mortality in the absence of HO-1.

Absence of cyclooxygenase-2 exacerbates hypoxia-induced pulmonary hypertension and enhances contractility of vascular smooth muscle cells.

BACKGROUND: Cyclooxygenase-2 (COX-2) is upregulated in pulmonary artery smooth muscle cells (PASMCs) during hypoxia and may play a protective role in the response of the lung to hypoxia. Selective COX-2 inhibition may have detrimental pulmonary vascular consequences during hypoxia. METHODS AND RESULTS: To investigate the role of COX-2 in the pulmonary vascular response to hypoxia, we subjected wild-type and COX-2-deficient mice to a model of chronic normobaric hypoxia. COX-2-null mice developed severe pulmonary hypertension with exaggerated elevation of right ventricular systolic pressure, significant right ventricular hypertrophy, and striking vascular remodeling after hypoxia. Pulmonary vascular remodeling in COX-2-deficient mice was characterized by PASMC hypertrophy but not increased proliferation. Furthermore, COX-2-deficient mice had significant upregulation of the endothelin-1 receptor (ET(A)) in the lung after hypoxia. Similarly, selective pharmacological inhibition of COX-2 in wild-type mice exacerbated hypoxia-induced pulmonary hypertension and resulted in PASMC hypertrophy and increased ET(A) receptor expression in pulmonary arterioles. The absence of COX-2 in vascular smooth muscle cells during hypoxia in vitro augmented traction forces and enhanced contractility of an extracellular matrix. Treatment of COX-2-deficient PASMCs with iloprost, a prostaglandin I(2) analog, and prostaglandin E(2) abrogated the potent contractile response to hypoxia and restored the wild-type phenotype. CONCLUSIONS: Our findings reveal that hypoxia-induced pulmonary hypertension and vascular remodeling are exacerbated in the absence of COX-2 with enhanced ET(A) receptor expression and increased PASMC hypertrophy. COX-2-deficient PASMCs have a maladaptive response to hypoxia manifested by exaggerated contractility, which may be rescued by either COX-2-derived prostaglandin I(2) or prostaglandin E(2).

Successful intraoperative electrophysiologic monitoring of the recurrent laryngeal nerve, a multidisciplinary approach: The Massachusetts Eye and Ear Infirmary monitoring collaborative protocol with experience in over 3000 cases.

BACKGROUND: Although intraoperative nerve monitoring (IONM) is utilized increasingly, the information on the related anesthesia technique is limited. This study presents an up-to-date clinical algorithm, including setup and troubleshooting of an IONM system, endotracheal tube placement, and anesthetic parameters. To our knowledge, this is the first interdisciplinary collaborative protocol for monitored neck surgery based on the published evidence and clinical experience. METHODS: The Departments of Otolaryngology Head and Neck Surgery, Anesthesiology, and Audiology collaboratively developed a protocol for IONM of the recurrent laryngeal nerve (RLN) based on published evidence and our experience with 3000 patients over a 16-year period. RESULTS: No complications related to monitoring or endotracheal tube placement were noted when the IONM protocol was implemented at Massachusetts Eye and Ear Infirmary (MEEI). The IONM protocol has proven to be vital in standardizing care and in avoiding intraoperative errors. CONCLUSION: An IONM system entails an anesthesiologist who understands the challenges posed by this technique; muscle relaxation must be minimized/eliminated to optimize IONM. © 2016 Wiley Periodicals, Inc. Head Neck 38: First-1494, 2016.

Distamycin A inhibits HMGA1-binding to the P-selectin promoter and attenuates lung and liver inflammation during murine endotoxemia.

BACKGROUND: The architectural transcription factor High Mobility Group-A1 (HMGA1) binds to the minor groove of AT-rich DNA and forms transcription factor complexes ("enhanceosomes") that upregulate expression of select genes within the inflammatory cascade during critical illness syndromes such as acute lung injury (ALI). AT-rich regions of DNA surround transcription factor binding sites in genes critical for the inflammatory response. Minor groove binding drugs (MGBs), such as Distamycin A (Dist A), interfere with AT-rich region DNA binding in a sequence and conformation-specific manner, and HMGA1 is one of the few transcription factors whose binding is inhibited by MGBs. OBJECTIVES: To determine whether MGBs exert beneficial effects during endotoxemia through attenuating tissue inflammation via interfering with HMGA1-DNA binding and modulating expression of adhesion molecules. METHODOLOGY/PRINCIPAL FINDINGS: Administration of Dist A significantly decreased lung and liver inflammation during murine endotoxemia. In intravital microscopy studies, Dist A attenuated neutrophil-endothelial interactions in vivo following an inflammatory stimulus. Endotoxin induction of P-selectin expression in lung and liver tissue and promoter activity in endothelial cells was significantly reduced by Dist A, while E-selectin induction was not significantly affected. Moreover, Dist A disrupted formation of an inducible complex containing NF-kappaB that binds an AT-rich region of the P-selectin promoter. Transfection studies demonstrated a critical role for HMGA1 in facilitating cytokine and NF-kappaB induction of P-selectin promoter activity, and Dist A inhibited binding of HMGA1 to this AT-rich region of the P-selectin promoter in vivo. CONCLUSIONS/SIGNIFICANCE: We describe a novel targeted approach in modulating lung and liver inflammation in vivo during murine endotoxemia through decreasing binding of HMGA1 to a distinct AT-rich region of the P-selectin promoter. These studies highlight the ability of MGBs to function as molecular tools for dissecting transcriptional mechanisms in vivo and suggest alternative treatment approaches for critical illness.

Reduction of nitric oxide synthase 2 expression by distamycin A improves survival from endotoxemia.

NO synthase 2 (NOS2) plays an important role in endotoxemia through overproduction of NO. Distamycin A (Dist A) belongs to a class of drugs termed minor-groove DNA binders, which can inhibit transcription factor binding to AT-rich regions of DNA. We and others have previously shown that AT-rich regions of DNA surrounding transcription factor binding sites in the NOS2 promoter are critical for NOS2 induction by inflammatory stimuli in vitro. Therefore, we hypothesized that Dist A would attenuate NOS2 up-regulation in vivo during endotoxemia and improve animal survival. C57BL/6 wild-type (WT) mice treated with Dist A and LPS (endotoxin) showed significantly improved survival compared with animals treated with LPS alone. In contrast, LPS-treated C57BL/6 NOS2-deficient (NOS2-/-) mice did not benefit from the protective effect of Dist A on mortality from endotoxemia. Treatment with Dist A resulted in protection from hypotension in LPS-treated WT mice, but not in NOS2-/- mice. Furthermore, LPS-induced NOS2 expression was attenuated in vivo (WT murine tissues) and in vitro (primary peritoneal and RAW 264.7 murine macrophages) with addition of Dist A. Dist A selectively decreased IFN regulatory factor-1 DNA binding in the enhancer region of the NOS2 promoter, and this IFN regulatory factor-1 site is critical for the effect of Dist A in attenuating LPS induction of NOS2. Our data point to a novel approach in modulating NOS2 expression in vivo during endotoxemia and suggest the potential for alternative treatment approaches for critical illness.