Leah D. Whigham

Selective inhibition of cyclooxygenase-2

Cyclooxygenase (COX), a key enzyme in the formation of prostanoids, is known to exist in two isoforms: an inducible enzyme (COX 2) and a constitutive from (COX 1). Both enzymes are inhibited by non-steroidal anti-inflammatory drugs (NSAID), but only marginal selectivity has thus far been reported. In this study, we report on a novel selective inhibitor of COX 2, CGP 28238 (6-(2,4-difluorophenoxy)-5-methyl-sulfonylamino-1-indanon e). Human washed platelets were used as a source of COX 1. For IL-1 stimulated rat mesangial cells we demonstrated the almost exclusive presence of COX 2 in western blot and mRNA analysis. Therefore these two model systems were chosen for selectivity testing. With an IC50 value of 15 nM, CGP 28238 blocked COX 2 activity in a similar concentration range to that of other potent NSAID such as indomethacin and diclofenac (IC50 = 1.17-8.9 nM). However, in contrast to these reference NSAIDs, CGP 28238 was at least 1000-fold less potent in inhibiting COX 1. Using other cell systems reported to express COX 1 or COX 2, we obtained a similar selectivity for COX 2. Thus, on the basis of our findings, CGP 28238 is a novel, highly potent and selective inhibitor of COX 2 and may be a lead compound for a new generation of potent anti-inflammatory drugs with an improved side-effect profile.

Skin and plasma carotenoid response to a provided intervention diet high in vegetables and fruit: uptake and depletion kinetics

BACKGROUND Objective biomarkers are needed to assess adherence to vegetable and fruit intervention trials. Blood carotenoids are considered the best biomarker of vegetable and fruit intake, but collecting blood is invasive and the analyses are relatively expensive for population studies. Resonance Raman spectroscopy (RRS) is an innovative method for assessing carotenoids in skin noninvasively. OBJECTIVE Our objective was to compare blood carotenoid concentrations with skin carotenoid assessments by RRS during a controlled feeding intervention. DESIGN Twenty-nine participants consumed low-carotenoid diets (6 wk, phases 1 and 3), a provided diet containing 6-cup equivalents (1046 g/d) of vegetables and fruit (8 wk, phase 2), and usual diet (final 8 wk, phase 4). RESULTS At baseline, skin and plasma total carotenoid values were correlated (r = 0.61, P < 0.001). Skin and plasma carotenoid values decreased (P < 0.001) 36% and 30%, respectively, from baseline to the end of phase 1 and then increased (P < 0.001) by >200% at the end of phase 2. Plasma carotenoids returned to baseline concentrations by the middle of phase 3 and skin carotenoid concentrations by the middle of phase 4. Skin carotenoid status predicted plasma values by using a mixed linear model including all time points (r = 0.72, P < 0.001), which indicates that changes in skin carotenoid status closely follow changes in plasma across a broad range of intakes. At the individual level, skin carotenoids predicted plasma values (r = 0.70, P < 0.001) over all time points. CONCLUSION Skin carotenoid status assessed by resonance Raman spectroscopy is a noninvasive, objective biomarker of changes in vegetable and fruit intake.

Strategies to Increase Vegetable or Reduce Energy and Fat Intake Induce Weight Loss in Adults

For obese individuals seeking to optimize health and well-being, healthy dietary strategies are important. Vegetables and fruits contribute to a healthy diet, and increased consumption may cause weight reduction by displacing foods high in energy and fat. The objective of this study was to determine if advising high vegetable (8 servings) and moderate fruit (2–3 servings) consumption would result in weight reduction in obese individuals. We compared this to advising a more traditional strategy of reducing daily energy intake by 500 kcal (2.1 MJ)/d and limiting energy from fat to ≤25%. A randomized study design was used. Subjects (age 21–50 y, n = 30/group) received food (2 meals + 1 snack/d, 5 d/wk) and education (2 group lessons/wk plus individual consultations as requested) for the first 3 mo. Weight and body composition were measured at baseline and after 3, 12, and 18 mo. Fasting serum lipid panel, insulin, glucose, hematocrit, and C-reactive protein were measured at baseline, 3, and 12 mo. Both groups lost weight after 3 mo (P = 0.0087 for high vegetable diet and P < 0.0001 for energy reduction diet), and the energy and fat reduction diet resulted in lower weight over time (P < 0.0001, treatment effect). Total cholesterol and cholesterol:HDL decreased after 3 mo in both groups (P ≤ 0.0061). Both strategies produced initial weight loss at 3 mo, but only the group following the caloric and fat reduction advice maintained weight loss at the 12- and 18-mo follow-up assessments. Nonetheless, the group following the high vegetable advice did not regain weight above baseline. In conclusion, traditional messages to reduce calories and fat are important, and increasing vegetable intake can assist individuals to maintain weight.

A meta-analysis of the effects of conjugated linoleic acid on fat-free mass in humans

Treatment of laboratory animals with a 50:50 mixture of c9,t11 and t10,c12 conjugated linoleic acid (CLA) results in fat loss and, to a smaller degree, fat-free mass (FFM) gain. In a previous meta-analysis, we found that CLA produced a fat loss, but that humans were not as responsive as mice. We performed a similar meta-analysis in the same 18 studies to test whether CLA increased FFM. Only placebo-controlled trials that measured body composition were included. We found that FFM increased during CLA treatment (0.3 +/- 0.7 kg; p = 0.05), but that the change did not display an effect of length of treatment (0.001 +/- 0.005 kg.week(-1); p = 0.8), or an effect of dosage (0.1 +/- 0.1 kg.g CLA(-1).day(-1); p = 0.3). We conclude that FFM does increase in humans during CLA treatment, but the onset of the increase is rapid and the total increase is small (<1%).

Prenatal androgen excess negatively impacts body fat distribution in a nonhuman primate model of polycystic ovary syndrome

Introduction:Prenatally androgenized (PA) female rhesus monkeys share metabolic abnormalities in common with polycystic ovary syndrome (PCOS) women. Early gestation exposure (E) results in insulin resistance, impaired pancreatic β-cell function and type 2 diabetes, while late gestation exposure (L) results in supranormal insulin sensitivity that declines with increasing body mass index (BMI).Objective:To determine whether PA females have altered body fat distribution.Design:Five early-treated PA (EPA), five late-treated PA (LPA) and five control adult female monkeys underwent somatometrics, dual-X-ray absorptiometry (DXA) and abdominal computed tomography (CT). Five control and five EPA females underwent an intravenous glucose tolerance test to assess the relationship between body composition and glucoregulation.Results:There were no differences in age, weight, BMI or somatometrics. LPA females had ∼20% greater DXA-determined total fat and percent body fat, as well as total and percent abdominal fat than EPA or control females (P⩽0.05). LPA females also had ∼40% more CT-determined non-visceral abdominal fat than EPA or control females (P⩽0.05). The volume of visceral fat was similar among the three groups. EPA (R 2=0.94, P⩽0.01) and LPA (R 2=0.53, P=0.16) females had a positive relationship between visceral fat and BMI, although not significant for LPA females. Conversely, control females had a positive relationship between non-visceral fat and BMI (R 2=0.98, P⩽0.001). There was a positive relationship between basal insulin and total body (R 2=0.95, P⩽0.007), total abdominal (R 2=0.81, P⩽0.04) and visceral (R 2=0.82, P⩽0.03) fat quantities in EPA, but not control females.Conclusions:Prenatal androgenization in female rhesus monkeys induces adiposity-dependent visceral fat accumulation, and late gestation androgenization causes increased total body and non-visceral fat mass. Early gestation androgenization induces visceral fat-dependent hyperinsulinemia. The relationship between the timing of prenatal androgen exposure and body composition phenotypes in this nonhuman primate model for PCOS may provide insight into the heterogeneity of metabolic defects found in PCOS women.

A Mobile Phone Food Record App to Digitally Capture Dietary Intake for Adolescents in a Free-Living Environment: Usability Study

Background Mobile technologies are emerging as valuable tools to collect and assess dietary intake. Adolescents readily accept and adopt new technologies; thus, a food record app (FRapp) may be a useful tool to better understand adolescents’ dietary intake and eating patterns. Objective We sought to determine the amenability of adolescents, in a free-living environment with minimal parental input, to use the FRapp to record their dietary intake. Methods Eighteen community-dwelling adolescents (11-14 years) received detailed instructions to record their dietary intake for 3-7 days using the FRapp. Participants were instructed to capture before and after images of all foods and beverages consumed and to include a fiducial marker in the image. Participants were also asked to provide text descriptors including amount and type of all foods and beverages consumed. Results Eight of 18 participants were able to follow all instructions: included pre- and post-meal images, a fiducial marker, and a text descriptor and collected diet records on 2 weekdays and 1 weekend day. Dietary intake was recorded on average for 3.2 (SD 1.3 days; 68% weekdays and 32% weekend days) with an average of 2.2 (SD 1.1) eating events per day per participant. A total of 143 eating events were recorded, of which 109 had at least one associated image and 34 were recorded with text only. Of the 109 eating events with images, 66 included all foods, beverages and a fiducial marker and 44 included both a pre- and post-meal image. Text was included with 78 of the captured images. Of the meals recorded, 36, 33, 35, and 39 were breakfasts, lunches, dinners, and snacks, respectively. Conclusions These data suggest that mobile devices equipped with an app to record dietary intake will be used by adolescents in a free-living environment; however, a minority of participants followed all directions. User-friendly mobile food record apps may increase participant amenability, increasing our understanding of adolescent dietary intake and eating patterns. To improve data collection, the FRapp should deliver prompts for tasks, such as capturing images before and after each eating event, including the fiducial marker in the image, providing complete and accurate text information, and ensuring all eating events are recorded and should be customizable to individuals and to different situations. Trial Registration Clinicaltrials.gov NCT01803997. http://clinicaltrials.gov/ct2/show/NCT01803997 (Archived at: http://www.webcitation.org/6WiV1vxoR).

Metabolic Evidence of Diminished Lipid Oxidation in Women With Polycystic Ovary Syndrome

Polycystic ovary syndrome (PCOS), a common female endocrinopathy, is a complex metabolic syndrome of enhanced weight gain. The goal of this pilot study was to evaluate metabolic differences between normal (n=10) and PCOS (n=10) women via breath carbon isotope ratio, urinary nitrogen and nuclear magnetic resonance (NMR)-determined serum metabolites. Breath carbon stable isotopes measured by cavity ring down spectroscopy (CRDS) indicated diminished (p<0.030) lipid use as a metabolic substrate during overnight fasting in PCOS compared to normal women. Accompanying urinary analyses showed a trending correlation (p<0.057) between overnight total nitrogen and circulating testosterone in PCOS women, alone. Serum analyzed by NMR spectroscopy following overnight, fast and at 2 h following an oral glucose tolerance test showed that a transient elevation in blood glucose levels decreased circulating levels of lipid, glucose and amino acid metabolic intermediates (acetone, 2-oxocaporate, 2-aminobutyrate, pyruvate, formate, and sarcosine) in PCOS women, whereas the 2 h glucose challenge led to increases in the same intermediates in normal women. These pilot data suggest that PCOS-related inflexibility in fasting-related switching between lipid and carbohydrate/protein utilization for carbon metabolism may contribute to enhanced weight gain.

Excess Gestational Weight Gain in Low-Income Overweight and Obese Women: A Qualitative Study.

OBJECTIVE Examine factors implicated in gestational weight gain (GWG) in low-income overweight and obese women. DESIGN Qualitative study. SETTING Community-based perinatal center. PARTICIPANTS Eight focus groups with women (black = 48%, white non-Hispanic = 41%, and Hispanic = 10%) in the first half (n = 12) and last half of pregnancy (n = 10) or postpartum (n = 7), 2 with obstetrician-gynecologists (n = 9). PHENOMENON OF INTEREST Barriers and facilitators to healthy eating and GWG within different levels of the Social Ecological Model: for example, intrapersonal, interpersonal, and organizational. ANALYSIS Coding guide was based on the Social Ecological Model. Transcripts were coded by 3 researchers for common themes. Thematic saturation was reached. RESULTS At an intrapersonal level, knowledge/skills and cravings were the most common barriers. At an interpersonal level, family and friends were most influential. At an organizational level, the Special Supplemental Nutrition Program for Women, Infants, and Children and clinics were influential. At the community level, lack of transportation was most frequently discussed. At a policy level, complex policies and social stigma surrounding the Special Supplemental Nutrition Program for Women, Infants, and Children were barriers. There was consensus that ideal intervention approaches would include peer-facilitated support groups with information from experts. Obstetrician-gynecologists felt uncomfortable counseling patients about GWG because of time constraints, other priorities, and lack of training. CONCLUSIONS AND IMPLICATIONS There are multilevel public health opportunities to promote healthy GWG. Better communication between nutrition specialists and obstetrician-gynecologists is needed.

Whole-body protein turnover response to short-term high-protein diets during weight loss: a randomized controlled trial

To determine whole-body protein turnover responses to high-protein diets during weight loss, 39 adults (age, 21±1 years; VO2peak, 48±1 ml kg−1 min−1; body mass index, 25±1 kg m2) were randomized to diets providing protein at the recommend dietary allowance (RDA), 2 × -RDA or 3 × -RDA. A 10-day weight maintenance period preceded a 21-day, 40% energy deficit. Postabsorptive (FASTED) and postprandial (FED) whole-body protein turnover was determined during weight maintenance (day 10) and energy deficit (day 31) using [1-13C]leucine. FASTED flux, synthesis and breakdown were lower (P<0.05) for energy deficit than weight maintenance. Protein flux and synthesis were higher (P<0.05) for FED than FASTED. Feeding attenuated (P<0.05) breakdown during weight maintenance but not energy deficit. Oxidation increased (P<0.05) between dietary protein levels and feeding stimulated oxidation, although oxidative responses to feeding were higher (P<0.05) for energy deficit than weight maintenance. FASTED net balance decreased between dietary protein levels, but in the FED state, net balance was lower for 3 × -RDA as compared with RDA and 2 × -RDA (diet-by-state, P<0.05). Consuming dietary protein at levels above the RDA, particularly 3 × -RDA, during short-term weight loss increases protein oxidation with concomitant reductions in net protein balance.

Comparison of combinations of drugs for treatment of obesity: body weight and echocardiographic status

Background:Obesity treatment with single drugs produces weight losses of about 8–10% of initial body weight. Few studies of combinations of drugs for treating obesity have been published. The combination of phentermine, an adrenergic agent, and fenfluramine, a serotonergic agent, (phen–fen) produced weight losses of about 15% of initial body weight. Fenfluramine is no longer available because it was associated with cardiac valve lesions. Phentermine–fluoxetine (phen–flu) has been proposed as an alternative for phen–fen.Objective:To compare the efficacy of treatment and prevalence of cardiac valve abnormalities on phen–flu vs phen–fen.Design:Retrospective chart review of all patients treated for at least 3 months with phen–flu (N=97) to a random sample of patients treated with phen–fen (N=98) in the Clinical Nutrition Clinic at the University of Wisconsin. Comparison of echocardiograms in all patients treated solely with phen–flu (N=21) to a random sample of patients treated with phen–fen (N=47), and to a group of subjects never treated with obesity drugs (N=26).Results:With last observation carried forward analysis (LOCF), at 6 months of treatment the phen–fen patients lost 12.6±0.6% of baseline weight and phen–flu patients lost 9.0±0.6% (P<0.001). With completers analysis, there were no significant differences in weight loss as a percent of baseline weight at 6 months (14.4±0.6 vs 13.3±0.9%). LOCF decreases in body mass index (BMI) at 6 months were −5.3 and −3.6 kg/m2 for phen–fen and phen–flu, respectively (P<0.001), and 6.2±0.3 vs 5.4±0.4 kg/m2, respectively, for the completers analysis (P – NS). Dropout rate at 6 months was higher in phen–flu subjects (44 vs 28%). In subjects without atherosclerosis of valves (presumably pre-existing), cardiac valve lesions occurred in eight of 38 phen–fen subjects and in none of 15 phen–flu subjects or 25 control subjects who had not been treated with drugs.Conclusions:The combination of phentermine and fluoxetine was not as effective as phen–fen, but was not associated with cardiac valve lesions. Longer term, larger scale studies of phen–flu are warranted.