Clifford Qualls

Background/Objectives: Previous studies have suggested that lifestyle intervention (LSI) therapies involving diet and exercise can modulate DNA methylation; however, whether this occurs in severely obese hypogonadal men undergoing weight loss from diet and exercise remains unclear. Methods: In this study, we investigated the effects of weight loss from diet and exercise on global DNA methylation as well as on the mRNA expression of specific demethylation enzymes, DNMT1, DNMT3A, and DNMT3B—in peripheral blood mononuclear cells (PBMCs) and DNA methylation markers in DNA of severely obese hypogonadal men. This is a secondary analysis of samples of severely obese (body mass index of ≥35 kg/m²) hypogonadal men undergoing weight loss from diet and exercise in addition to an aromatase inhibitor (anastrozole) or placebo for a total of 12 months. Results: LSI therapy significantly reduced global DNA methylation and 5-methylcytosine (5-mC) levels, decreased DNMT1, DNMT3A, and DNMT3B (p < 0.05) mRNA levels and markedly decreased CEBPα, FTO, and PPARγ mRNA expression. The reduction in global methylation was independent of aromatase inhibitor use. Conclusions: In summary, our findings suggest that LSI induces epigenetic modifications in leukocytes, possibly through the regulation of DNMT gene expression. Future studies are warranted to clarify the mechanistic pathways linking lifestyle-induced epigenetic alterations to metabolic health outcomes.

Purpose: The purpose of this study is to identify associative factors of vision threatening complications of diabetic retinopathy (DR), including proliferative diabetic retinopathy (PDR) and diabetic macular edema (DME) in a New Mexican patient population with Type 2 diabetes featuring a high proportion of Hispanic and American Indian patients. Methods: In a retrospective, case‐control, cross‐sectional study, we performed both univariate and multivariate logistic regression testing of systemic factors for association among patients with nonproliferative diabetic retinopathy (NPDR) without DME, NPDR with DME, and PDR. We also used receiver operating characteristic (ROC) curves to understand how reliable glycemic control is at predicting DR end‐complications. Results: Among our 584 patients, 172 were diagnosed with NPDR without DME, 293 with PDR, and 119 with NPDR with DME. A total of 25% of patients with NPDR without DME had poor glycemic control (HbA1c levels of ≥ 9.0%). Similarly, 10% of PDR and 7% of NPDR with DME patients had good glycemic control (HbA1c levels ≤ 7.0%), despite their advanced eye disease. For patients with PDR, we identified significant independent associations with microalbuminuria and macroalbuminuria, Hispanic ethnicity, duration of diabetes, and the use of insulin and beta‐blocker medications. In NPDR with DME patients, significant associations were noted with microalbuminuria and macroalbuminuria, HbA1c, GFR, and beta‐blocker medications. The hemoglobin A1c level was not significantly associated with PDR but was significantly associated with NPDR with DME. The ROC curve analysis indicated poor predictability for the HbA1c model concerning the presence of PDR (AUC = 0.640) and NPDR with DME (AUC = 0.640). Conclusions: This study suggests that glycemic exposure alone is not a good enough predictor of DR progression. Other diabetic microvascular complications such as nephropathy, in addition to race/ethnicity with differing genetic associative factors, may also play a role in the multifactorial progression through DR phenotypes.

Grape seed procyanidin extract (GSE) is widely used to promote cardiovascular health and has purported anti-inflammatory properties. Chronic inflammation in the lungs caused by environmental toxins such as tobacco smoking plays a pivotal role in lung cancer development. In a modified phase I lung cancer chemoprevention study conducted in heavy active and former smokers using leucoselect phytosome (LP), a standardized grape seed procyanidin extract complexed with soy phospholipids to enhance bioavailability, three months of LP treatment favorably modulated a variety of surrogate endpoint biomarkers, including markers of cell proliferation. In this correlative study, we further analyzed the effects of LP on cytochrome P450 3A4 (CYP3A4) activities by comparing the endogenous conversions of cortisol and cortisone to 6-beta-hydroxycortisol and 6-beta-hydroxycortisone, respectively, before and after LP treatment and the anti-inflammatory effects of LP in the lung microenvironment of these participants by comparing a profile of inflammatory cytokines and chemokines in matched pre- and post-treatment bronchoalveolar lavage (BAL) fluids. LP treatment did not significantly alter CYP3A4 activity, and three months of LP treatment significantly decreased tumor necrosis factor (TNF), C-C Motif Chemokine Ligand 3 (CCL3) and granzyme B in BAL fluids. Furthermore, post-LP-treatment BAL fluids significantly reduced migration/invasion of various human lung neoplastic cells in vitro. Our findings support the anti-inflammatory effects of GSE/LP in the lung microenvironment and its potential utility for reducing cancerizing forces, as well as driving forces for other common respiratory diseases such as chronic obstructive pulmonary disease and asthma, in the lungs of heavy former and active smokers.

Cortisol is secreted by the human adrenal cortex and circulates in plasma as free or protein-bound cortisol. Corticosteroid binding globulin (CBG) and albumin are the principal binding proteins (BPs) for cortisol in human plasma. Plasma concentrations of total cortisol (sum of protein-bound and free cortisol) are typically measured in vitro. Determination of free cortisol adds clinical and diagnostic value to total cortisol concentration. However, direct measurement of free cortisol concentrations involves laborious separation methods, limiting clinical utility. The development and application of physiologic protein-ligand binding models and equations provide an alternative approach to assessment of free cortisol concentrations in vitro. In this chapter, we introduce a matrix notation to represent relevant mass action and mass conservation equations. The matrix notation is also used to summarize and compare several contemporary models of interest, including cubic, quadratic, and quartic polynomial equations. Second, we introduce Feldmans equations for competitive ligand-protein binding interactions, which are represented by matrices for multiple ligands and multiple BPs, including illustrative 2 2 matrix; we also discuss iterative solution strategies for coupled polynomial equations. Third, we develop a theorem for albumin-cortisol binding and review related assumptions that have been used to simplify polynomial equations and their equilibrium solutions.

Context Estimated rates of cortisol elimination and appearance vary according to the model used to obtain them. Generalizability of current models of cortisol disposition in healthy humans is limited. Objective Development and validation of a realistic, mechanistic model of cortisol disposition that accounts for the major factors influencing plasma cortisol concentrations in vivo (Model 4), and comparison to previously described models of cortisol disposition in current clinical use (Models 1-3). Methods The four models were independently applied to cortisol concentration data obtained for the hydrocortisone bolus experiment (20 mg) in two clinical groups: healthy volunteers (HV, n = 6) and CBG-deficient (n = 2). Model 4 used Fick’s first law of diffusion to model free cortisol flux between vascular and extravascular compartments. Pharmacokinetic parameter solutions for Models 1-4 were optimized by numerical methods, and model-specific parameter solutions were compared by RM-ANOVA. Models and respective parameter solutions were compared by mathematical and simulation analyses, and an assessment tool was used to compare performance characteristics. Results Cortisol half-lives differed significantly between models (all P < 0.001) with significant model-group interaction (P = 0.02). In comparative analysis, Model 4 solutions yielded significantly reduced free cortisol half-life, improved fit to experimental data (both P < 0.01), and superior model performance. Conclusions The proposed 4-compartment diffusion model (Model 4) is consistent with relevant experimental observations and met the greatest number of empiric validation criteria. Cortisol half-life solutions obtained using Model 4 were generalizable between HV and CBG-deficient groups and bolus and continuous modes of hydrocortisone infusion.

Background: Emerging data suggest that type 2 diabetes mellitus (T2DM) is associated with increased risk for fractures despite relatively normal or increased bone mineral density (BMD). Furthermore, it is now known that decreased bone turnover mainly due to reduced bone formation is the hallmark for bone disease in T2DM. Whether glucose control is important in generating this impairment in bone metabolism remains unknown and to what extent it would reflect this abnormality is undetermined. The purpose of our study is to identify Hemoglobin A1c (A1c) level threshold by which reduction in bone turnover begins. Method: Baseline data from 217 men between age of 35–65 who were participants in 2 clinical trials conducted at the Michael DeBakey VA Medical Center and the New Mexico VA Health Care System were analyzed. A1c was measured by high performance liquid chromatography, testosterone and estradiol measured by liquid chromatography/mass spectrometry (LC/MS). Bone turnover markers (Osteocalcin [OC],C-telopeptide of type 1 Collagen [CTx]) and sclerostin were measured by enzyme-linked immunosorbent assay. Bone mineral density was assessed by dual energy X-ray absorptiometry. Patients were grouped into 4 categories based of A1c values (%) (group 1:7). Simple correlations were assessed by simple regression analysis and group comparisons among the different A1c categories were performed by analysis of variance (ANOVA). Results: The mean age of the participants was 55±9 years old with mean BMI of 36.15±6.44 kg/m². Participants mean A1c was 6.1±1.5%. Simple correlation analysis showed a significant negative correlation between A1C and OC (r=-0.32, pConclusion: Our data analysis showed breakpoint A1c level of 7% or greater is associated with lower bone turnover irrespective of medication use in patients with T2DM.

Background We sought to determine whether prenatal supplementation with the omega-3 fatty acids eicosapentaenoic acid (EPA) or docosahexaenoic acid (DHA) would increase markers of insulin sensitivity in maternal or cord blood compared with placebo supplementation. A secondary aim was to evaluate the association of serum EPA and DHA fractions with adiponectin, leptin and the adiponectin:leptin ratio (ALR). We hypothesized that omega-3 fatty acid supplementation would increase markers of insulin sensitivity in maternal and umbilical cord plasma.Methods We analyzed stored plasma samples collected from a prior 3-arm prospective, double-blinded, randomized controlled trial in which 126 women with singleton pregnancies between 12- and 20-weeks gestation were randomized to receive: 1) an EPA-rich fish oil supplement, 2) a DHA-rich fish oil supplement, or 3) a soy oil placebo. Maternal venous blood samples were collected at 12-20 weeks gestation (before supplementation) and at 34-36 weeks gestation. At delivery, cord blood was collected. Samples were analyzed using enzyme-linked immunosorbent assay kits to quantify leptin and adiponectin levels which were utilized to calculate the ALR, a proxy measure for insulin sensitivity.Results We found no difference in adiponectin, leptin, and the ALR between the treatment and placebo groups at baseline, after supplementation, or in umbilical cord blood. In regression analyses, higher maternal serum DHA fraction was associated with increased ALR before (p = 0.01) and after (p = 0.04) DHA supplementation. There was no association of EPA fraction with any measure of insulin sensitivity. Cord blood DHA fraction was significantly associated with cord plasma leptin (p = 0.02). Early pregnancy BMI was significantly associated with maternal leptin levels at baseline and in late pregnancy (p < 0.001) and was inversely associated with the ALR (p < 0.001). The ALR decreased significantly between the early and late pregnancy visits (p < 0.001). Pregnancy weight gain was inversely associated with the ALR (P. <0.02). Conclusions EPA- and DHA- rich fish oil supplementation had no effect on plasma markers of insulin sensitivity. However, maternal serum DHA fraction was significantly associated with markers of insulin sensitivity.

Background We sought to determine whether prenatal supplementation with the omega-3 fatty acids eicosapentaenoic acid (EPA) or docosahexanoic acid (DHA) would increase markers of insulin sensitivity in maternal or cord blood compared with placebo supplementation. A secondary aim was to evaluate the association of serum EPA and DHA fractions with adiponectin, leptin and the adiponectin:leptin ratio (ALR). We hypothesized that omega-3 fatty acid supplementation would increase markers of insulin sensitivity in maternal and umbilical cord plasma.Methods We analyzed stored plasma samples collected from a prior 3-arm prospective, double-blinded, randomized controlled trial in which 126 women with singleton pregnancies between 12- and 20-weeks’ gestation were randomized to receive: 1) an EPA-rich fish oil supplement, 2) a DHA-rich fish oil supplement, or 3) a soy oil placebo. Maternal venous blood samples were collected at 12-20 weeks gestation (before supplementation) and at 34-36 weeks gestation. At delivery, cord blood was collected. Samples were analyzed using sandwich enzyme-linked immunosorbent assay kits to quantify leptin and adiponectin levels which were utilized to calculate the ALR, a proxy measure for insulin sensitivity.Results We found no difference in adiponectin, leptin, and the ALR between the treatment and placebo groups at baseline, after supplementation, or in umbilical cord blood. In regression analyses, higher maternal serum DHA fraction was associated with increased ALR before (p = 0.01) and after (p = 0.04) DHA supplementation. There was no association of EPA fraction with any measure of insulin sensitivity. Cord blood DHA fraction was significantly associated with cord plasma leptin (p = 0.02). Early pregnancy BMI was significantly associated with maternal leptin levels at baseline and in late pregnancy (p < 0.001) and was inversely associated with the ALR (p < 0.001). The ALR decreased significantly between the early and late pregnancy visits (p < 0.001). Pregnancy weight gain was inversely associated with the ALR (P. <0.02). Conclusions EPA- and DHA- rich fish oil supplementation had no effect on plasma markers of insulin sensitivity. However, maternal serum DHA fraction was significantly associated with markers of insulin sensitivity.