Prenatal treatment differentially affects offspring weight and weight gain over the lifespan.

<p>(A) During the lactation period, ETOH offspring gained less weight than did other groups, and this decreased weight gain was significant in female ETOH offspring in comparison with MD controls (p<0.05). (B) During the juvenile period (P21-P35), both male and female ETOH offspring gained significantly more weight in comparison to other groups (M: p<0.02; F: p<0.01). (C) During adolescence and adulthood (P35-P105), ETOH offspring gained less weight than other groups (M: p<0.0001 vs all other groups; F: p<0.05 vs. MD), despite their greater weight gain during the juvenile period. Values are mean ± SEM of more than 20 per sex*treatment group. * p<0.05 vs. H2O, ** p<0.01 vs. H2O, *** p<0.001 vs. H2O, # p<0.05 vs. isocaloric control (MD or MCT, as indicated by bar), using mixed linear factorial analysis of variance, followed by slice-effect ANOVAs with <i>a priori</i> hypotheses allowing for planned comparisons.</p

Prenatal calories, but not PAE, differentially impairs oral glucose tolerance in offspring.

<p>An oral glucose tolerance test (OGTT) was performed in 17-week-old PAE and control offspring (A-D), and again after they consumed a high-fat diet for four weeks. Blood glucose concentrations and insulin levels were assessed at times following an intragastric glucose bolus. (A) Male offspring receiving added gestational calories (MCT, MD, and ETOH) had higher blood glucose levels in response to oral glucose challenge as compared with H2O males. Males receiving PAE or isocaloric MD had significantly greater area-under-the-curve (AUC) values (inset) as compared with H2O-control males, and their AUC values did not differ from each other, suggesting that glucose clearance was reduced in those isocaloric groups. Males that received isocaloric MCT had a trend to elevated AUC compared with those that received H2O. (B) In these same males, PAE did not affect blood insulin levels as compared with H2O and isocaloric MD or MCT controls at either baseline fast or at 15 minutes post-glucose bolus. (C) PAE did not affect glucose clearance in female offspring as compared with H2O or isocaloric MD or MCT controls, nor did PAE alter the AUC for glucose clearance (inset). (D) PAE did not affect blood insulin levels in female offspring as compared with H2O and isocaloric controls at baseline fast and at fifteen minutes post-glucose bolus. (E) Consumption of a high-fat diet for four weeks elevated fasting blood glucose in PAE and control males. PAE did not further affect blood glucose or AUC values (inset) during the OGTT, as compared with controls. (F) In these males, high-fat diet consumption both elevated fasting insulin levels and their insulin levels at fifteen minutes post-glucose bolus. These levels were not further affected by PAE or prenatal treatment. (G) Consumption of a high-fat diet for four weeks elevated fasting blood glucose levels in PAE and control females. PAE did not further affect blood glucose or AUC values (inset) during the OGTT, as compared with controls. (H) In these same females, high-fat diet feeding elevated both fasting insulin and blood insulin levels at fifteen minutes post glucose bolus in all treatment groups. These values were not further affected by PAE or prenatal treatment. Values are mean ± SEM of 8–12 offspring per sex*treatment group. * p<0.05 vs. H2O, ** p < 0.01 vs. H2O, # p<0.05 vs. MCT, <i>t</i> p<0.1 vs. H2O, using mixed linear factorial analysis of variance, followed by slice-effect ANOVAs with <i>a priori</i> hypotheses allowing for planned comparisons.</p

PAE does not alter whole body metabolism in adult male and female mice.

<p>Male and female offspring were evaluated for food intake and whole body metabolism using environmental chambers. The individual’s ability to adapt its metabolism and food intake to diet composition was assessed by sequential feeding of chow diet (3 days), low fat diet (10%, 3 days), and high fat diet (60%, 4 days). Body composition was assessed at the beginning and end of each diet challenge using NMR. (A) Body weight at the beginning and end of each diet challenge did not differ by treatment in males (left panel) and was reduced in ETOH females only during chow feeding. (B) The percentage of body size as fat mass increased on the low-fat and high-fat diets, but was not affected by PAE or other treatments in male and female offspring. (C) Cumulative water intake (measured hourly) was not affected by prenatal treatment in either sex. (D) Cumulative food intake increased on the low-fat and high-fat diets compared with chow diet, but did not differ with respect to prenatal treatment. (E) Hourly food intake showed similar patterns across the dark/light cycle in males, and was significantly affected by prenatal treatment in females, as detailed in the text. (F) Oxygen consumption (VO2, ml/h/kg) showed diurnal variation consistent with food consumption pattern and activity, and was unaffected by prenatal treatment in either sex. (G) The calculated respiratory exchange ratio (RER) showed diurnal variation consistent with the pattern of food intake and, as expected, was increased with the low-fat diet and decreased with the high-fat diet. RER was not affected by PAE or other prenatal treatments in either sex. Shaded bars represent the dark cycle when mice are more active. Values are mean ± SEM of 8–10 offspring per sex*treatment group; SEMs are omitted from panels C-G for clarity. Blue, H2O; Green, MD; Black, MCT; Red, ETOH.</p

Isocaloric treatment, but not PAE, alters fat mass in adult male but not female mice.

<p>(A) Body composition (% lean mass, % fat mass) was quantified in control and alcohol-treated offspring using DXA. At age 17 weeks, male offspring had similar body weights and lean mass content, expressed as a percentage of body mass, regardless of prenatal treatment. However, ETOH males had significantly greater fat mass compared with MD males. (B) At age 17 weeks, female offspring had similar body weights, lean mass content, and fat mass content, expressed as a percentage of body mass, regardless of prenatal treatment. (C) In a separate cohort of males dissected for tissue collection at age 17 weeks, prenatal treatment did not affect total body weight, brown fat mass, and gastrocnemius mass, the latter two expressed as a percentage of body mass. However, male offspring in all three added-calorie groups (MCT, MD, ETOH) had a significantly decreased inguinal fat pad mass in comparison with H2O males (p<0.05). (D) In females, there were no ETOH or treatment effects upon body weight, or fat pad and gastrocnemius mass as a percentage of body weight. Values are mean ± SEM of 10–12 offspring per sex*treatment group. * p<0.05 vs. H2O, # p<0.05 vs. MD, using mixed linear factorial analysis of variance, followed by slice-effect ANOVAs with <i>a priori</i> hypotheses allowing for planned comparisons.</p

High-fat diet feeding does not unmask a unique adiposity phenotype in PAE offspring.

<p>Body composition was assessed by NMR in adult control and PAE mice, before and after they consumed a high-fat diet for 4 weeks. (A) High-fat diet intake caused significant weight gain in all male offspring, regardless of prenatal treatment. The weight gain percentage in ETOH males was higher than in MD males, but this was only a trend (p<0.07). (B) Chow-fed ETOH females weighed significantly less than all other groups at age 17 weeks. Consumption of a high-fat diet for four weeks increased female body weight regardless of prenatal treatment. However, ETOH and MD females weighed significantly less than their H2O counterparts (p<0.03), and their weight gain percentage was also decreased compared with H2O controls (p<0.03). (C) Fat mass significantly increased in males fed a high-fat diet, and this gain, as a percentage of weight, was not differentially affected by prenatal treatment. (D) Fat mass increased in females fed a high-fat diet, and this gain, as a percentage of body weight, was significantly less in MD and ETOH females (vs. H2O females, p<0.05). (E) Lean mass in male offspring was unaffected by high-fat diet or prenatal treatment. (F) Lean mass in female offspring was unaffected by high-fat diet or prenatal treatment. Values are mean ± SEM of 8–10 offspring per sex*treatment group. * p<0.05 vs. H2O, # p<0.05 vs. MCT, using mixed linear factorial analysis of variance, followed by slice-effect ANOVAs with <i>a priori</i> hypotheses allowing for planned comparisons.</p

Experimental design for metabolic assessment in alcohol-exposed offspring.

<p>(A) Primary study design. Prenatal gavage treatment of timed pregnancies began at E12.5 and continued daily through E17.5 using three controls (H2O, isocaloric MCT, or isocaloric MD) or a 3 g/kg ETOH dose. Offspring developed without interference until seventeen weeks of age, when male/female offspring pairs within each litter were randomized into one of three assessment arms: (1) Tissue collection at 17 weeks (n = 8–10 mice per treatment*sex group); (2) Body composition and metabolic cage assessment, followed by a thirty-day high fat (60%) diet challenge and oral glucose tolerance testing (n = 10–12 mice per treatment*sex group); (3) oral and intraperitoneal glucose tolerance testing, followed by blood pressure assessment (n = 8–10 mice per treatment*sex group). (B) A separate group of pregnant dams were gavaged with either 4.5 g/kg dose or isocaloric MD control as in (A). Body composition of offspring was assessed at 4, 6, 8, and 16 weeks of age. Fasting blood glucose was evaluated at 16 weeks.</p

Gestational alcohol exposure does not adversely affect maternal growth and litter size.

<p>(A) Maternal weight during the gavage period. Due to a partial randomization of treatment, ETOH dams were initially significantly heavier than other dams at E0.5 (p<0.05) and during the dosing period (p<0.01). However, treatment did not affect weight gain during the gavage period (inset, p>0.1), suggesting a lack of effect by alcohol. (B) Litter parameters at E17.5. Total maternal weight gain from E0.5 to E17.5 did not significantly differ across treatment groups, although ETOH dams gained the most weight. This weight gain likely represented the significant increase in ETOH litter size, because weight gain per pup was not affected by treatment. Values are mean ± SEM of 12 dams per treatment group. * p<0.05 vs. H2O, using mixed linear factorial analysis of variance, followed by slice-effect ANOVAs with <i>a priori</i> hypotheses allowing for planned comparisons.</p

Echocardiography of 13–16 months old wild-type and <i>Polg</i>^D257A/D257A mice on control and calorie-restricted diets.

<p>Echocardiography of 13–16 months old wild-type and <i>Polg</i>^D257A/D257A mice on control and calorie-restricted diets.</p

Stimuli-Responsive Polymer Coatings for the Rapid and Tunable Contact Transfer of Plasmid DNA to Soft Surfaces

We report the design and characterization of thin polymer-based coatings that promote the contact transfer of DNA to soft surfaces under mild and physiologically relevant conditions. Past studies reveal polymer multilayers fabricated using linear poly(ethylene imine) (LPEI), poly(acrylic acid) (PAA), and plasmid DNA promote contact transfer of DNA to vascular tissue. Here, we demonstrate that changes in the structure of the polyamine building blocks of these materials can have substantial impacts on rates and extents of contact transfer. We used two hydrogel-based substrate models that permit identification and manipulation of parameters that influence contact transfer. We used a planar gel model to characterize films having the structure (cationic polymer/PAA/cationic polymer/plasmid DNA)x fabricated using either LPEI or one of three poly(β-amino ester)s as polyamine building blocks. The structure of the polyamine influenced subsequent contact transfer of DNA significantly; in general, films fabricated using more hydrophilic polymers promoted transfer more effectively. This planar model also permitted characterization of the stabilities of films transferred onto secondary surfaces, revealing rates of DNA release to be slower than rates of release prior to transfer. We also used a three-dimensional hole-based hydrogel model to evaluate contact transfer of DNA from the surfaces of inflatable catheter balloons used in vascular interventions and selected a rapid-transfer coating for proof-of-concept studies to characterize balloon-mediated contact transfer of DNA to peripheral arterial tissue in swine. Our results reveal robust and largely circumferential transfer of DNA to the luminal walls of peripheral arteries using inflation times as short as 15 to 30 s. The materials and approaches reported here provide new and useful tools for promoting rapid, substrate-mediated contact transfer of plasmid DNA to soft surfaces in vitro and in vivo that could prove useful in a range of fundamental and applied contexts

Body weight, skeletal muscle mass, and testis weight of mitochondrial mutator mice.

<p>Body weights (A), vastus lateralis (VL) and gastrocnemius (GN) muscle mass (B) of 13–16 months old +/+, +/D257A, and D257A/D257A males (left) and females (right) under control diet or calorie restricted conditions. (C) Testes weights of 10–18 months old +/+, +/D257A, and D257A/D257A males under control diet or calorie restricted conditions. ‡P < 0.05 control diet vs CR diet within genotype. n = 8–16. *P < 0.05 +/+ vs D257A/D257A within diet. #P < 0.05 +/+ vs +/D257A within diet. §P < 0.05 +/D257A vs D257A/D257A within diet. +/+ = <i>wild-type</i>, +/D257A = <i>Polg</i>^+/D257A, and D257A/D257A = <i>Polg</i>^D257A/D257A.</p