Attacks on minoritized communities and increasing awareness of the societal causes of health disparities have combined to highlight deep systemic inequities. In response, academic health centers have prioritized justice, equity, diversity, and inclusion (JEDI) in their strategic goals. To have a sustained impact, JEDI efforts cannot be siloed; rather, they must be woven into the fabric of our work and systematically assessed to promote meaningful outcomes and accountability. To this end, the University of Pittsburgh’s Institute for Clinical Research Education assembled a task force to create and apply a rubric to identify short and long-term JEDI goals, assess the current state of JEDI at our Institute, and make recommendations for immediate action. To ensure deep buy-in, we gathered input from diverse members of our academic community, who served on targeted subcommittees. We then applied a three-step process to ensure rapid forward progress. We emerged with concrete actions for priority focus and a plan for ongoing assessment of JEDI institutionalization. We believe our process and rubric offer a scalable and adaptable model for other institutions and departments to follow as we work together across academic medical institutions to put our justice, equity, diversity, and inclusion goals into meaningful action.

Twelve evidence-based profiles of roles across the Clinical and Translational Science (CTS) workforce and two patient profiles were developed by CTS Personas collaborators in 2019 as part of the CTSA Program National Center for Data to Health (CD2H). Based on feedback received from the community, CTS Personas team members collaborated to produce five additional Personas to broaden representation of the CTS workforce and enhance the existing portfolio. This paper presents the rationale and methodology used in the latest CTS Personas initiative. This work also includes an implementation scenario incorporating multiple Personas. Using the new National Institutes of Health’s (NIH) Data Management and Sharing Policy as an example, we demonstrate how administrators, researchers, support staff, and all CTS collaborators can use the Personas to respond to this new policy while considering the needs of service providers and users, CTS employees with short- and long-term needs, and interdisciplinary perspectives.

Identifying and disseminating actionable intelligence is a challenging task that requires thoughtful planning. The National Center for Advancing Translational Sciences instituted the Common Metrics Initiative with the goal of evaluating the Clinical and Translational Science Awards (CTSA) Programs using a standard set of metrics. Initially managed by Tufts University, the Center for Leading Innovation and Collaboration (CLIC) at the University of Rochester began leading this initiative in 2017. In directing this work, CLIC created a framework for communicating and disseminating data insights. Insights to Inspire emerged from the need to share strategies and lessons learned to improve metric performance at the local level to a network of 60+ academic research institutions. Insights to Inspire employs a mixed methods approach for translating data into actionable intelligence. A series of blogs, webinars, and webcasts were designed to communicate metric-specific strategies used by individual sites to the broader CTSA consortium. A dissemination plan to expand the reach beyond metric stakeholders utilized focused communications including social media channels, network newsletters, and presentations at national meetings. This framework serves as a blueprint for other national evaluation programs interested in a systematic approach to using data insights for continuous improvement.

Despite understanding its impact on organizational effectiveness, practical guidance on how to train translational team (TT) leaders is lacking. Previously, we developed an evolutionary learning model of TT maturation consisting of three goal-directed phases: (1). team assembly (Formation); (2). conducting research (Knowledge Generation); and (3). dissemination and implementation (Translation). At each phase, the team acquires group-level knowledge, skills, and attitudes (KSAs) that enhance its performance. Noting that the majority of team-emergent KSAs are promoted by leadership behaviors, we examine the SciTS literature to identify the relevant behaviors for each phase. We propose that effective team leadership evolves from a hierarchical, transformational model early in team Formation to a shared, functional leadership model during Translation. We synthesized an integrated model of TT leadership, mapping a generic “functional leadership” taxonomy to relevant leadership behaviors linked to TT performance, creating an evidence-informed Leadership and Skills Enhancement for Research (LASER) training program. Empirical studies indicate that leadership behaviors are stable across time; to enhance leadership skills, ongoing reflection, evaluation, and practice are needed. We provide a comprehensive multi-level evaluation framework for tracking the growth of TT leadership skills. This work provides a framework for assessing and training relevant leadership behaviors for high-performance TTs.

OBJECTIVES/SPECIFIC AIMS: As pediatric care becomes more concentrated in large urban hospitals, smaller rural hospitals with reduced pediatric care capacity may opt to transfer pediatric patients to higher levels of care even if the patient has a condition that is manageable in a general ED. Up to 20-40% of pediatric transfers are considered avoidable, placing a burden on the patient, their family and the health care system. The aim of this study is to determine the association between pediatric readiness (as measured by the National Pediatric Readiness Project score) and risk of interfacility transfer. We hypothesized that emergency departments with higher measures of pediatric readiness would be less likely to transfer pediatric patients to another facility. METHODS/STUDY POPULATION: The most recent and complete National Pediatric Readiness Project data were from 300 California hospitals in 2012. These data include the overall pediatric readiness score, presence of inter-facility guidelines (written protocols on patients needing care not available at the hospital), presence of interfacility agreements (written agreements with other hospitals regarding transfer patients), and other variables on the facility’s capacity to care for pediatric patients. We linked these hospital data with patient encounter data from the California Office of Statewide Health Planning and Development Emergency Department (ED) Database (OSHPD) using hospital name and zip code. To be eligible for the study, the patient must have a documented ED encounter and be less than 18 years old during the encounter. A patient was considered a transfer patient if they were transferred to a separate general hospital, children’s hospital, federal health care facility, rehabilitation facility, Critical Access Hospital or psychiatric hospital. Chi-square tests and t-tests were used for descriptive statistics. For non-normal data, we used the non-parametric Kruskall-Wallis test. We also used logistic regression to compare the odds of transfer between comparison groups. Statistical analyses were conducted in the R environment version 3.4.4. RESULTS/ANTICIPATED RESULTS: In 2012, there were 2,604,723 pediatric ED encounters, of which 10,966 resulted in a transfer (0.4%). Transferred patients on average were older (15 vs. 5 years, p < 0.001) and more likely to be female (58.6% vs 46.6%, p < 0.001). The transfer group originated from hospitals with a lower median pediatric readiness score (76.0 vs 78.3, p < 0.001). Patients were less likely to be transferred if they were seen at a hospital with written guidelines with transfer protocols (OR 0.89, 95% CI 0.83—0.95, p < 0.001). Patients were more likely to be transferred if they were seen at a hospital with written interfacility agreements with other hospitals (OR 1.17, 95%CI 1.10—1.25, p < 0.001). We anticipate, that even with more sophisticated multilevel statistical models, pediatric readiness scores will remain associated with odds of transfer. DISCUSSION/SIGNIFICANCE OF IMPACT: These preliminary data suggest that hospitals with higher levels of pediatric readiness and written guidelines with transfer protocols are less likely to transfer pediatric patients. There may be actionable policy and procedural items that a hospital could enact to lower the rate of transfer patients. Future analyses will include more complex statistical modeling to adjust for confounders, will include inpatient data, and will compare the risk of potentially avoidable transfers between hospitals with varying levels of pediatric readiness.

Health systems currently underutilize systematic reviews. Here, we describe a proof of concept project designed to augment the standard systematic review process by presenting qualitative information as a companion to a review on deprescribing interventions. We conducted a thematic analysis of semi-structured interviews with Veterans Health Administration clinicians and Veterans to describe first-hand experiences of engaging in the deprescribing process. Qualitative findings were incorporated into an interactive, web-based product designed to supplement the systematic review report. Preliminary evaluation suggests that integration of narratives as a companion to systematic reviews is of interest to frontline clinicians, researchers, and health system administrators.

OBJECTIVES/GOALS: Metabolic syndrome (MetSyn) is a risk for World Trade Center-Lung Injury (WTC-LI; defined as developing FEV1<lower limit of normal [LLN]). Metabolic health is a modifiable disease risk factor. We propose to characterize how time-dependent covariates of MetSyn are longitudinally associated with WTC-LI. METHODS/STUDY POPULATION: WTC-particulate exposed firefighters, consented, with pre-9/11 FEV1 LLN (N = 5,746). Data assessed from last pre-9/11 till August 1, 2017. Longitudinal MetSyn characteristics were assessed using 3 models: i. A linear mixed effect model to assess the effect size of longitudinal MetSyn and its components on longitudinal FEV1% predicted as an outcome; ii. a time-dependent Cox regression to assess the associations of MetSyn to time of onset of WTC-LI; iii. a novel, partially linear single index regression model with repeatedly measured MetSyn to assess their joint effects and delineate their relative contribution on the longitudinal lung function in the WTC-FDNY cohort. RESULTS/ANTICIPATED RESULTS: In Model I, BMI 30 kg/m2 had the largest effect size compared to ever-smoking, with −2.524 (95%CI: −2.708,−2.340) compared to −1.681(−2.325,−1.038) respectively. Having MetSyn, defined as 3/5 risk factors, had an effect size of −2.319(−2.526,−2.112). In Model II, hazards of triglycerides 150mg/dL were highest at 1.497(1.336, 1.677), followed by BMI 30 kg/m2 at 1.406(1.256, 1.575), and HDL<40mg/dL 1.355(1.176-1.561), compared to ever-smoking (1.201, p = 0.002). Having high exposure to PM by being present in the morning of 9/11 was a significant covariate only in Model II investigating HDL<40mg/dL or triglycerides 150mg/dL. Model III The proposed methods will be applied to our cohort study. DISCUSSION/SIGNIFICANCE OF IMPACT: MetSyn is both a predictor and concurrent marker of WTC-LI. The single index model can not only reduce dimensionality of the covariates, but also provides efficient estimates of the joint MetSyn effects, allowing linear or nonlinear effects. Future studies will investigate dietary intervention as a potential disease-modifying factor. CONFLICT OF INTEREST DESCRIPTION: NA, nothing to disclose.

OBJECTIVES/SPECIFIC AIMS: Cardiovascular disease (CVD) is the leading cause of death among US adults and its prevalence is increasing, despite efforts to identify, and address risk factors. Post-traumatic stress disorder (PTSD) has been identified as a potential risk factor for CVD, though the results to date have focused on male veterans with combat-related PTSD. To our knowledge, there are no prospective analyses/reports among older community-dwelling adults following Hurricane Katrina. The purpose of this study was to explore the link between PTSD associated with Hurricane Katrina and incident CVD among elderly adults using data from the Cohort Study of Medication Adherence among Older Adults (CoSMO). METHODS/STUDY POPULATION: PTSD associated with Katrina and incident CVD events were assessed among 2075 hypertensive participants age ≥65 who were enrolled in a managed care organization in southeastern Louisiana. Baseline surveys were conducted between August 2006 and September 2007. Baseline surveys were conducted between August 2006 and September 2007. PTSD was assessed using the civilian PTSD CheckList (PCL-17) and 2 cut-off points, ≥37 and ≥44, for primary and secondary analyses, respectively. Participants were followed through February 2011 for a composite CVD outcome of MI, stroke, CHF, or CVD death. Multivariable logistic regression was performed with 13 covariates identified in bivariate analysis: age, sex, race, marital status, education, hypertension knowledge, comorbidities, number of antihypertensive medication classes, dissatisfaction with healthcare, reduced medications due to cost, number of visits to healthcare provider in last year, depression, and coping. RESULTS/ANTICIPATED RESULTS: Participants were 59.8% female and 30.4% black, with a mean age of 75 years. The prevalence of PTSD using the primary and secondary cut points was 6.1% and 4.2%, respectively. In total, 240 (11.5%) participants had a CVD event during a median 3.8 year follow-up. After multivariable adjustment, the odds ratios and 95% confidence intervals (CI) for CVD event for the primary and secondary analyses were 1.90 (95% CI: 1.17, 3.09) and 3.74 (95% CI: 2.05, 6.81), respectively. DISCUSSION/SIGNIFICANCE OF IMPACT: PTSD was associated with an increased risk of incident CVD events among elderly adults. This finding from a prospective cohort study supports earlier reports suggesting PTSD is an independent risk factor for CVD. To our knowledge, this association has not been previously reported among a cohort of elderly community-dwelling adults. This study included hypertensive, elderly, insured participants living in southeastern Louisiana following Hurricane Katrina and may not be generalizable to all people with PTSD. Strengths of this study include its longitudinal design, the identification of incident CVD, the diversity of the study population with respect to gender, race and CV risk, and reduced confounding due to access to care and insurance status. Future research is needed to confirm this finding in other populations and to assess if efforts to minimize the impact of PTSD following disasters reduce CVD risk and premature CVD events among older adults.

OBJECTIVES/GOALS: During and after cancer treatment, cancer-related fatigue (CRF) is a debilitating symptom reported by up to 80% of cancer patients Our understanding of the pathology underlying CRF is limited. Preliminary RNA sequencing data suggest that increased levels of KIR3DL1, the natural killer cell (NK) immunoglobulin-like receptor 3DL1a, may be associated with CRF. METHODS/STUDY POPULATION: Fatigue was measured using the Functional Assessment of Chronic Illness Therapy-Fatigue (FACT-F). Functional validation of the NK cell finding was performed from whole blood obtained from fatigued and non-fatigued subjects. NK cells were isolated from freshly collected whole blood using a human NK cell isolation kit based on CD56 microbead positive selection. NK cell function was assessed using the NK cell direct cytotoxicity assay. Briefly, isolated NK cells were co-cultured in a 2:1 ratio with calcein AM-labelled K562 cells, which are NK cell-sensitive due to the very low MHCI expression. NK cell-mediated cytotoxicity was assessed with Cytation 1 Cell Imaging Multi-Mode reader. Flow cytometric protocols were used to examine NK subset differences between the fatigued and non-fatigued groups. RESULTS/ANTICIPATED RESULTS: NK cells isolated from the fatigued group exhibited decreased cytotoxicity at 12.28% compared to NK cells isolated from non-fatigued controls at a mean of 40.6% cytotoxicity. Flow cytometry analysis revealed a decrease in the CD56dim CD16bright population in the fatigued group (87.1% of CD56+CD4- cells) compared to the control (91.4% of CD56+CD4- cells). Furthermore, there was a decrease in NKG2A expression in mature NK cells (CD56dim CD16bright) isolated from the fatigued group compared to the non-fatigued group. DISCUSSION/SIGNIFICANCE OF IMPACT: Results from the pilot study suggest that there was a decrease in NK cell cytotoxicity in the fatigued group. In addition, there may be a shift in NK cell subpopulations associated with fatigue. Findings from this pilot study suggest that impaired NK cell function may be associated with CRF pathogenesis.

OBJECTIVES/GOALS: Our preclinical data demonstrate that the principal effects of nilotinib, a multi-tyrosine kinase inhibitor, in models of neurodegeneration is clearance of misfolded proteins via autophagy. Here we aimed to evaluate the effects of nilotinib on microRNAs in the cerebrospinal fluid of Parkinson’s disease patients. METHODS/STUDY POPULATION: Cerebrospinal fluid (CSF) was collected as part of an open label phase I (NCT02281474) (n = 12, 300 mg nilotinib taken orally once daily for 6 months), and a phase II randomized, double-blind, placebo-controlled study (NCT02954978) (n = 75, randomized 1:1:1 into placebo, 150 mg or 300 mg nilotinib taken orally once daily for 12 months). RNA was isolated from CSF and Indexed sequencing libraries were prepared from total RNA plus miRNA. Next generation whole-genome sequencing (single-end 1x75 bp, 25 million raw reads per sample) was performed to identify miRNAs significantly differentially expressed (fold-change ≥ 2, Benjamini-Hochberg FDR p-value ≤ 0.05 or Empirical Bayes FDR ≤ 0.05) with treatment compared to baseline. RESULTS/ANTICIPATED RESULTS: Next generation whole-genome sequencing of microRNAs in the CSF demonstrated that nilotinib significantly increases microRNAs that specifically regulate expression of autophagy and ubiquitination genes in individuals with Parkinson’s disease. In the open label phase I, samples, 28 microRNAs found to regulate autophagy and ubiquitination genes, were significantly altered with treatment (Benjamini-Hochberg FDR p-value ≤ 0.05). In the phase II randomized, double-blind, placebo-controlled study samples, we verified several of those 28 candidate microRNAs had been significantly deferentially expressed with treatment (Empirical Bayes FDR p-value ≤ 0.05). DISCUSSION/SIGNIFICANCE OF IMPACT: Our data provide robust evidence that nilotinib’s effects on misfolded protein clearance is via autophagy and CSF miRNA sequencing is a valid biomarker of nilotinib’s effects in a definitive phase III study to investigate nilotinib in Parkinson’s and other neurodegenerative diseases. CONFLICT OF INTEREST DESCRIPTION: Charbel Moussa is listed as an inventor on several Georgetown University patents for the use of tyrosine kinase inhibitors as a treatment for neurodegenerative diseases

OBJECTIVES/GOALS: There is no objective, biologic tool to detect concussion or guide clinical management. We previously showed that saliva microRNA (miRNA) levels differ in children with concussion and may predict symptom duration. The purpose of this study was to validate the utility of saliva miRNA and define longitudinal trends during the recovery period. METHODS/STUDY POPULATION: We collected concussion symptom burden (SCAT-5), cognitive performance (DANA), balance measures (ClearEdge), and saliva from 150 children (7-21 years) with concussion over 5 time-points: 0-2, 3-6, 7-14, 15-29, and 30-60 days post-injury. Saliva miRNA levels within the 443 concussion samples were quantified with RNA sequencing and compared to 218 samples from age- and sex-matched controls (healthy and post-exercise participants). Non-parametric ANOVA assessed RNA levels across time-points, and between concussions/controls. Machine learning was used to build logistic regression algorithms differentiating concussions/controls, and symptomatic/recovered concussion participants. Relationships between miRNAs and concussion phenotypes were explored with Spearman’s Rank correlations. RESULTS/ANTICIPATED RESULTS: Fifteen miRNAs differed across control and concussion participants (FDR < 0.05). Within concussion participants, all 15 miRNAs trended back toward control levels by 30-60 days post injury. A regression algorithm employing 6 of the 15 miRNAs differentiated control and concussion participants with an area under the curve (AUC) of 0.78 in a training set (n = 244) and 0.84 in a naïve test set (n = 24). Similarly, 6 miRNAs were able to differentiate symptomatic (SCAT-5 symptom score > 7) and asymptomatic concussion participants with an AUC of 0.73 in a training set (n = 219) and 0.76 in a naïve test set (n = 44). Furthermore, 5 miRNAs showed significant (R > 0.3; FDR < 0.05) associations with subjective and/or objective measures of concussion-related symptoms. DISCUSSION/SIGNIFICANCE OF IMPACT: Saliva miRNAs levels are altered in children with concussion, and display predictable longitudinal trends following injury. Saliva miRNA measurement represents a non-invasive, objective tool that could be rapidly assessed to provide biologic evidence for clinicians managing pediatric concussion. CONFLICT OF INTEREST DESCRIPTION: I serve as a paid consultant and scientific advisory board member for Quadrant Biosciences, who has funded a portion of this work and licensed the findings from the Penn State College of Medicine.

OBJECTIVES/SPECIFIC AIMS: We evaluated the accuracy of diagnosis and procedure codes to identify acute urinary retention (AUR) due to lack of existing validation studies. METHODS/STUDY POPULATION: We performed a cross-sectional validation study at a single medical institution in the emergency department (ED) and outpatient Urology Clinic in men ≥45 years. International Classification of Diseases, Ninth Revision, Clinical Modification (ICD-9-CM) diagnosis codes 788.20, 788.21, 788.29 for urinary retention and Current Procedural Terminology, Fourth Edition (CPT-4) codes 51701, 51702, 51703 for urinary catheterization were used to identify men with potential AUR. Four algorithms using ICD-9-CM and CPT-4 codes were compared against medical record review. Sensitivity, specificity, positive predictive value, negative predictive value, and area under the curve were calculated for both the ED and Urology Clinic. RESULTS/ANTICIPATED RESULTS: A total of 333 treated and released men in the ED were identified using facility billing data, and 245 men using physician billing data in the Urology Clinic were identified using the codes for urinary retention or urinary catheterization. Of the 4 algorithms, any ICD-9-CM diagnosis code for urinary retention was the preferred algorithm with a sensitivity and specificity of 0.95 and 0.91 using ED facility billing data and a sensitivity and specificity of 0.95 and 0.58 using Urology physician billing data. DISCUSSION/SIGNIFICANCE OF IMPACT: Use of the ICD-9-CM diagnosis codes for urinary retention performed well at identifying AUR in the ED. This study provides justification to use urinary retention diagnosis codes (specifically 788.20 and 788.29) in future studies to identify AUR using administrative data, especially in the ED.

Clinical and translational science is vitally dependent on the nation’s underlying health-care policies and programs. In a reciprocal fashion, data generated by clinical and translational research can inform both health policy and health-care delivery. It is important, therefore, to rate health reform proposals comprehensively on a set of criteria that reflect the broad goals of reform, including the potential impact on clinical and translational science and medical education. I propose that the criteria include achieving universal coverage, reducing administrative costs, retaining one’s chosen primary care physician, encouraging care coordination, empowering physicians, freeing industry from choosing and administering health plans, providing choice of specialists and hospitals, providing patient education, preventing patient overuse of services, rationalizing resource allocation, encouraging competition, limiting government’s role, supporting medical education, training, and research, and freeing industry to make personnel decisions based on business criteria rather than the impact on health-care costs to the company. I discuss the rationale for each element and offer a rating of current proposals relative to a proposal previously made.

OBJECTIVES/GOALS: High cholesterol is among the major causes of cardiometabolic complications. People with high cholesterol have about twice the risk of heart disease as people with lower levels. Approximately every 40 seconds, an American will have a heart attack. Costs related to Heart Attack exceed 60 Billion USA dollars per year. Renin-Angiotensin-Aldosteron System (RAAS) is implicated in the genesis of coronary heart disease and in the perpetuation of heart failure. Angiotensin-Converting Enzyme inhibitors (ACE-I) have emerged as the treatment of choice for patients with all degrees of heart failure. Many clinical trials (Consensus, 1987; Save 1990) provide the evidence that ACE-I preserves cardiac function, prevents cardiovascular death, myocardial infarction & stroke and limit remodeling after myocardial infarction. However, there are still controversies in cardiology and a debate over cardioprotection is continuing:

• Do ACE Inhibitors have unique properties, beyond their antihypertensive effect?

• Can we protect the heart during hypercholesterolemia?

• In which way hypercholesterolemia affects mitochondria bioenergetics?

• How does ramipril affect mitochondrial bioenergetics during the course of experimental hypercholesterolemia?

Objectives/Goals were: To evaluate the mitochondrial actions of chronically administered ramipril (non-SH-containing ACE inhibitor) in cholesterol-fed rabbits by determining the influence of ramipril on:

• myocardial oxygen consumption (State 4, State 3), Respiratory Control Ratio (RCR), and adenosine diphosphate - oxygen index (ADP/O) and

• oxidative stress biomarkers

METHODS/STUDY POPULATION: Animal treatments. In the course of twelve-week the male Chinchilla rabbits (n = 10/group) received once a day a single dose, in group: A - sunflower oil (control animals); B – atherogenic 2% hypercholesterolemic diet; C - atherogenic diet and ramipril (1 mg/kg) and D - ramipril (1 mg/kg) only. Animals were terminated in accordance with the U.K. “Animals (Scientific Procedures) Act.” Isolation of mitochondria - Mitochondria from rabbit heart were isolated by tissue digestion (trypsin), fractionation and differential centrifugation. Mitochondrial respiratory functional measures (State 4 - Basal; State 3 - ADP-stimulated respiration, RCR and ADP/O) and biochemical markers of oxidative damage (the nitrite level) were measured polarographically (Clark electrode, YSI, USA) and spectrophotometrically, respectively, in isolated heart mitochondrial suspensions. Statistics - All results are reported as means ± SD. Comparisons between ramipril treated and control animals were performed by unpaired t-test or one-way ANOVA with a Tukey adjustment for multiple comparisons. A P value < 0.05 was considered significant for all tests. RESULTS/ANTICIPATED RESULTS: Plasma cholesterol levels: After a period of 12 weeks

• Plasma cholesterol levels in control rabbits (A) were low (1.36 ± 0.23 mmol/l).

• Cholesterol-fed rabbits became hypercholesterolemic and their plasma total cholesterol level was higher even than 10 mmol/l. The level of total cholesterol in the high-cholesterol-diet group was significantly increased compared with the level in the normal-diet group (p < 0.01).

• In the high-cholesterol-diet group treated with ramipril (C), the plasma cholesterol level was not affected by the drug ramipril (10.54 ± 1.31 mmol/l). ACE-I ramipril did not infuence the concentration of total cholesterol.

• Plasma cholesterol levels in group D were low (1.46 ± 0.29 mmol/l).

• Atherogenic 2% cholesterol diet (B) caused a decline in mitochondrial function (in both, State 3 and 4) (−25%).

• Mitochondria from group C animals (treatment with ramipril along with 2% cholesterol diet) exhibited higher State 3 respiratory rates compared with group B.

• Mild inhibition of mitochondrial respiration was recorded in group D, in both respiratory states (V4&V3).

• In cholestrerol-supplemented hearts myocardial oxygen consumption was markedly reduced (State 4 and State 3) compared to controls. Administration of high-cholesterol diet decreased not only the respiratory activity of rabbit heart mitochondria (RHM), but also the sensitivity of respiratory chain to ADP (ADP/O), while concomitantly caused an increase in nitrite production. Possible explanation: high-fat diet affects the fluidity of mitochondrial membrane – Electron transport chain (ETC) may be damaged, and unable to support high rates of respiration (e.g. substantial cytochrome c could be lost).

• Administration of ACE-I ramipril along with cholesterol diet partially abolish reduction in MQO2 and improved coupling efficiency (ADP/O). Possible explanation: reduced coupling efficiency means the coupling mechanism itself is altered (e.g. the respiratory complexes slip and pump fewer protons than normal and less ATP is produced per oxygen consumed. Ramipril partially improved coupling efficiency and increased the amount of ATP per oxygen consumed.

• RCR - No significant difference between the groups were found. High RCR indicates good function (a high capacity for substrate oxidation and ATP turnover). Low RCR usually indicates dysfunction. However, there is no absolute RCR value that is diagnostic of dysfunctional mitochondria, because values are substrate- and tissue-dependent.

• NO• exerts metabolic control over mitochondrial respiration. Group B: The lowered state 3-respiration in heart mitochondria seems to contribute to the increased NO production, and elevated nitrite level.

• In a system as complex as OXPHOS, conclusions about overall efficiency must involve measurements of: mito membrane potential, proton transport, ATP synthesis and modular kinetic analisys.