Comparative Phylogenetics of Papilio Butterfly Wing Shape and Size Demonstrates Independent Hindwing and Forewing Evolution.

The complex forces that shape butterfly wings have long been a subject of experimental and comparative research. Butterflies use their wings for flight, camouflage, mate recognition, warning, and mimicry. However, general patterns and correlations among wing shape and size evolution are still poorly understood. We collected geometric morphometric measurements from over 1400 digitized museum specimens of Papilio swallowtails and combined them with phylogenetic data to test two hypotheses: 1) forewing shape and size evolve independently of hindwing shape and size and 2) wing size evolves more quickly than wing shape. We also determined the major axes of wing shape variation and discovered that most shape variability occurs in hindwing tails and adjacent areas. We conclude that forewing shape and size are functionally and biomechanically constrained, whereas hindwings are more labile, perhaps in response to disruptive selective pressure for Batesian mimicry or against predation. The development of a significant, re-usable, digitized data resource will enable further investigation on tradeoffs between flight performance and ecological selective pressures, along with the degree to which intraspecific, local-scale selection may explain macroevolutionary patterns. [Batesian mimicry; Lepidoptera; geometric morphometrics; museum specimens.].

Preweaning food intake influences the adiposity of young adult baboons.

The hypothesis that preweaning nutrition influences adult fat cell number and adiposity was tested in baboons. Newborn baboons were fed Similac formulas with caloric densities of 40.5 kcal (underfed), 67.5 kcal (fed normally), and 94.5 kcal (overfed) per 100 g formula. From weaning (16 wk) until necropsy at 5 yr of age all baboons were fed the same diet. At necropsy, fat cell number and fat cell size in 10 fat depots were measured. Female baboons overfed as infants had markedly greater fat depot mass, primarily because of fat cell hypertrophy, than normally fed or underfed females. Overfed male baboons had a greater fat mass in 4 of 10 depots compared with males underfed or fed normally as infants. Underfeeding did not affect body weight, nor adipose mass of either sex. The results show that infant food intake does not have a major influence on the fat cell number of young adult baboons.

Effects of dietary cholesterol on adipose tissue lipoprotein lipase in the baboon.

The effects of infant diet (breast milk or formula containing 2, 30 or 60 mg/dl cholesterol) and subsequent dietary cholesterol (0.02, 1.0 or 1.7 mg/kcal) and fat (saturated or unsaturated) on heparin-releasable lipolytic activity from omental adipose tissue was estimated from 99 baboons of 5-8 years of age. This lipase activity was characterized as lipoprotein lipase based on salt inhibition and apolipoprotein C-II activation. Lipoprotein lipase activity released from adipose tissue by heparin was significantly (P less than 0.002) lower in high cholesterol-fed baboons than in those fed low cholesterol. Most of this difference was due to impaired long-term heparin release of lipoprotein lipase. Adipose tissue lipoprotein lipase increased with increasing fat cell size regardless of diet, but there was no effect of diet on adipocyte size. There were no significant effects of infant cholesterol intake nor adult saturated or unsaturated fat on lipoprotein lipase activity. Adult baboons breast fed as infants had lower adipose tissue lipoprotein lipase activity (P less than 0.07) than adults fed formula as infants.

An assessment of the role of protein kinase and zymogen granule phosphorylation during secretion by the rat exocrine pancreas.

1. The levels of protein kinase activity and zymogen granule phosphorylation were studied in the adult rat during stimulus-coupled secretion in vitro. 2. The specific activity of protein kinase associated with intact zymogen granules was 11 pmol [32P]phosphate transferred to histone per min per mg protein. Most of this activity was recovered in purified granule membranes. 2. The addition of 10(-6) M cyclic AMP to a mixture of zymogen granules and the postmicrosomal supernatant resulted in a 5-fold increase in protein kinase activity associated with zymogen granules. The adsorbed activity was eluted from granules by 0.15 M NaCl. Cyclic GMP did not promote protein kinase binding to isolated granules. 4. Incubation of tissues with carbachol (10(-5) M), pancreozymin (0.1 unit/ml), caerulein (10(-8) M) or dibutyryl cyclic AMP (2.10(-4) M) between 2.5 and 60 min did not increase the levels of protein kinase activity in isolated zymogen granules above control values. 5. Protein phosphorylation of zymogen granule membranes and granule content was not detectable in tissues incubated with carbachol, pancreozymin-C-octapeptide, or caerulein. 6. These results suggest that neither the phosphorylation of zymogen granule membrane protein nor the adsorption of protein kinase activity to zymogen granules is an obligatory step in secretion.

Influence of breast and formula feeding on hepatic concentrations of apolipoprotein and low-density lipoprotein receptor mRNAs.

We tested the hypothesis that breast and formula feeding differentially affect hepatic mRNA concentrations for LDL receptor (LDL-R) and apolipoproteins A-I, B and E in infant baboons during the preweaning period. The mRNA concentrations were measured in liver biopsies obtained prior to weaning at 14 weeks from 43 baboons that were either breast-fed (n = 17) or fed formulas with a high (n = 12) or low (n = 14) polyunsaturated/saturated (P:S) fat ratio. Breast-fed baboons had 99% higher LDL-R mRNA concentrations compared with infants fed formulas, but there were no differences among breast and formula-fed baboons in mRNA concentrations of apolipoproteins A-I, B or E. The fatty acid P:S ratio of the formulas did not affect hepatic LDL-R or apolipoprotein mRNA concentrations. These results suggest that breast-feeding increases LDL-R gene expression even though breast milk is higher in cholesterol and saturated fat compared with formulas.

Glycoprotein synthesis in the adult rat pancreas. IV. Subcellular distribution of membrane glycoproteins.

Zymogen granule membranes from the rat exocrine pancreas displays distinctive, simple protein and glycoprotein compositions when compared to other intracellular membranes. The carbohydrate content of zymogen granule membrane protein was 5-10-fold greater than that of membrane fractions isolated from smooth and rough microsomes, mitochondria and a preparation containing plasma membranes, and 50-100-fold greater than the zymogen granule content and the postmicrosomal supernate. The granule membrane glycoprotein contained primarily sialic acid, fucose, mannose, galactose and N-acetylglucosamine. The levels of galactose, fucose and sialic acid increased in membranes in the following order: rough microsomes less than smooth microsomes less than zymogen granules. Membrane polypeptides were analyzed by polyacrylamide gel electrophoresis in sodium dodecyl sulfate. The profile of zymogen granule membrane polypeptides was characterized by GP-2, a species with an apparent molecular weight of 74 000. Radioactivity profiles of membranes labeled with [3H]glucosamine or [3H]leucine, as well as periodic acid-Schiff stain profiles, indicated that GP-2 accounted for approx. 40% of the firmly bound granule membrane protein. Low levels of a species similar to GP-2 were detected in membranes of smooth microsomes and the preparation enriched in plasma membranes but not in other subcellular fractions. These results suggest that GP-2 is a biochemical marker for zymogen granules. Membrane glycoproteins of intact zymogen granules were resistant to neuraminidase treatment, while those in isolated granule membranes were readily degraded by neuraminidase. GP-2 of intact granules was not labeled by exposure to galactose oxidase followed by reduction with NaB3H4. In contrast, GP-2 in purified granule membranes was readily labeled by this procedure. Therefore GP-2 appears to be located on the zymogen granule interior.

Effect of premature birth and survival on hepatic thyroxine 5'-monodeiodinase activity in baboons.

Plasma thyroid hormones and hepatic 5'-monodeiodinase type I (5'-MDI) activity were measured in a primate model of premature birth and survival. When prematurely delivered at 140 days (term gestation is 184 days), infant baboons developed hyaline membrane disease, had no surge in T3 and T4, temporarily developed hypothyroxinemia, and had extremely low T3 concentrations during the first 40 h of life. After 4 days, both plasma T4 and T3 levels progressively increased, but were still considerably lower at 16 days compared to those in normal term infants. Hepatic 5'-MDI activity was measured in the presence of dithiothreitol in fetal, premature, and infant baboons. Fetal baboons at 140 and 161 days gestation had 80% less 5'-MDI activity than term infants, but at 178-180 days gestation, near term, fetal hepatic 5'-MDI activity increased to levels similar to those in young adults. Interruption of in utero development by premature birth resulted in no change in hepatic 5'-MDI activity after 24 h, but within 6 days after delivery, hepatic 5'-MDI had significantly increased to levels observed in fetal baboons near term. Kinetic analysis revealed that fetal and premature 5'-MDI had different maximum velocities and similar apparent Km values. There was no significant difference in hepatic total, protein, or nonprotein sulfhydryl groups between 1- and 10-day-old premature (140 days gestation) baboons. These results suggest that premature birth does not limit the postnatal maturation of hepatic 5'-MDI activity.

Effect of energy intake on postprandial plasma hormones and triglyceride concentrations in infant female baboons (Papio species).

We previously reported that female baboons overfed during infancy were not fatter at weaning, but developed hypertrophic obesity after puberty. To clarify the mechanisms of this dietary effect on adipocyte hypertrophy, we determined the effects of infant overfeeding on preweaning plasma hormone and triglyceride levels and their relationship with fat cell volume at weaning (19 weeks of age). Newborn female baboons from 3 sires and 24 dams were fed either 280 kilojoules (normally fed; n = 12) or 395 kilojoules (overfed; n = 10) per 100 g Similac formula for 18 weeks. Both formulas contained 9.2%, 43.1%, and 48.5% of calories as protein, carbohydrate, and fat, respectively. During the first 9 weeks, overfed infants had significantly higher fasting and postprandial insulin, total T3, and free T3 concentrations; lower cortisol levels; and lower excretion of urinary 17-hydroxycorticosteroids (17-OHCS) than normally fed infants. These effects were no longer significant at 17-18 weeks. Infant diet did not influence fasting and postprandial plasma triglyceride levels, and fat cell volume was not influenced by energy intake. However, fat cell volume was positively associated with postprandial triglyceride concentrations and inversely associated with postmeal nadir cortisol levels. These results demonstrate that infant overfeeding initiates early alterations in insulin, T3, free T3, and cortisol, but these effects persist only as long as there is a significant increase in energy intake.

A quality assurance program for human immunodeficiency virus seropositivity screening of dried-blood spot specimens.

Epidemiologic projects have been implemented for using dried-blood spot (DBS) specimens routinely collected on filter paper from neonates as a seroepidemiologic resource to monitor seroprevalence of human immunodeficiency virus (HIV) among childbearing women. To ensure the quality of the data base produced from the national and other epidemiologic surveys, a quality assurance program was developed to interact with all the neonatal screening laboratories. The focus of the Centers for Disease Control's quality assurance program for HIV seropositivity testing of neonatal blood specimens is to maintain a national program to produce, certify, and provide external quality control materials as DBSs on filter paper, to monitor the filter paper matrix, to operate an external performance surveillance program, and to provide other special and consultative services. The dried-blood control and surveillance materials are certified by rigorous testing for accuracy, stability, and homogeneity. Preliminary results from the first performance evaluation of screening laboratories indicated excellent performance.

Early determinants of adult metabolic regulation: effects of infant nutrition on adult lipid and lipoprotein metabolism.

In a series of experiments over the past 20 years, we have demonstrated long-term deferred effects of infant nutrition, particularly breast- as compared with formula-feeding and overfeeding as compared with normal or underfeeding, on serum HDL-cholesterol concentrations, adiposity, and atherosclerosis in the baboon, a large nonhuman primate. Low HDL-cholesterol levels and obesity are associated with accelerated progression of atherosclerosis and with increased risk of coronary heart disease in humans. We have observed other deferred effects of infant nutrition on bile acid metabolism, enzyme activities, and water and electrolyte balance, some of which may be physiologically related to HDL-cholesterol levels or to adiposity. The occurrence of these deferred effects suggests that infant nutrition may program other metabolic systems for life, and that these effects may contribute to other chronic diseases of adults. Although our understanding of the mechanisms by which infant diet regimens affect adult metabolism is meager, it is important to identify these mechanisms because they are likely to provide valuable clues to the causes and ultimately may contribute to the long-range prevention of those diseases.

In vivo and in vitro development of visceral adipose tissue in a nonhuman primate (Papio species).

We determined the development of the omental fat depot in a cross-sectional study of 242 baboons from birth to mature adulthood. The triglyceride content of the omentum increased during preweaning (birth to 4 months) and adolescence (2 to 5 years) and was associated with an increase in both fat cell number and size. Between weaning and 2 years of age omentum triglyceride mass decreased as a result of decreasing fat cell size, but fat cell number remained constant. After adolescence and up to 13 years of age, omental triglyceride mass and fat cell volume were stable, but fat cell number increased slightly in female baboons. We determined the in vitro potential of omental stromal vascular (S-V) cells from baboons at different stages of development to differentiate in a serum-free medium. Both the proportion of omental S-V cells that accumulated cytoplasmic lipid droplets and the induction of glycerol phosphate dehydrogenase (GPDH) activity were increased to the greatest degree in the presence of 1-methyl-3-isobutylxanthine, 2.0 nmol/L triiodothyronine (T3), 0.85 mumol/L insulin, and 1.0 mumol/L cortisol. Omental S-V cells from preweaning and adolescent baboons had a greater differentiation rate, GPDH activity, and triglyceride accumulation compared with cells from postweaned infants and mature adults. In summary, most of the growth of the baboon omentum occurs during the preweaning and pubertal periods of life, and omental S-V cells isolated from animals during these periods retain a greater potential to differentiate in vitro.

The role of cholesterol absorption and hepatic cholesterol content in high and low responses to dietary cholesterol and fat in pedigreed baboons (Papio species).

Selective breeding has produced baboon families with low and high plasma cholesterol responses to dietary cholesterol and fat. We used 12 high- and 12 low-responding (mainly in low-density lipoprotein [LDL] cholesterol) pedigreed baboons to determine whether cholesterol absorption and hepatic cholesterol concentration are associated with these responses. We measured cholesterol absorption first on the chow diet, which was low in cholesterol and fat, and after 3 and 13 weeks on the challenge diets, which contained 0.45 mg cholesterol/kcal and 40% of calories as either coconut oil or corn oil. Plasma, lipoprotein, and hepatic cholesterol concentrations were measured 1 week after cholesterol absorption measurements. High-responding baboons had higher percentage cholesterol absorption than low-responding baboons on both chow and challenge diets, regardless of the type of dietary fat. Both high and low responders had higher percentage cholesterol absorption with corn oil than with coconut oil. High responders also had higher hepatic cholesterol concentrations than low responders on chow and after consuming the challenge diets for 4 weeks. After consuming the challenge diets for 14 weeks, low responders fed coconut oil had hepatic cholesterol levels equal to those of high responders, while low responders fed corn oil continued to have low hepatic cholesterol levels. Thus, percentage cholesterol absorption is consistently higher in high-responding baboons regardless of diet, but hepatic cholesterol concentration varies with duration of challenge and type of fat. The results suggest that both cholesterol absorption and hepatic cholesterol concentration regulate cholesterolemic responses to diet, but by different mechanisms.

Effect of dietary lipids on plasma activity and hepatic mRNA levels of cholesteryl ester transfer protein in high- and low-responding baboons (Papio species).

We compared the effects of dietary cholesterol, type of fat (coconut oil v corn oil), and phenotype (low low-density lipoprotein [LDL] response v high LDL response) on the plasma activity and hepatic mRNA levels of cholesteryl ester transfer protein (CETP). In a crossover design, eight high- and eight low-LDL-responding baboons were fed a series of diets with increasing amounts of cholesterol (0.05, 0.15, 0.45, and 1.35 mg/kcal) with either coconut oil or corn oil. All diets were fed for 7 weeks each. plasma and lipoprotein cholesterol concentrations, plasma lecithin:cholesterol acyltransferase (LCAT) and CETP activity, and hepatic mRNA levels for CETP and apolipoprotein (apo) A-I were measured after 6 weeks on each diet. Data were analyzed in two steps, ie, the effect of the initial change from chow to 0.05 mg cholesterol with each fat and the effect of the stepwise increase in cholesterol from 0.05 to 1.35 mg/kcal with each fat. High-responding baboons, as expected, showed a more pronounced increment in plasma LDL cholesterol at all dietary cholesterol levels, particularly with coconut oil as the dietary fat. Plasma high-density lipoprotein 2 (HDL2) and HDL3 cholesterol increased as dietary cholesterol increased on both the coconut and corn oil diets, with a greater increase in high-responding baboons than in low-responding baboons. The stepwise increase in dietary cholesterol increased plasma LCAT activity in both high- and low-responding baboons fed the coconut oil diet, but not in those fed the corn oil diet. Dietary cholesterol, regardless of type of fat, increased plasma CETP activity.(ABSTRACT TRUNCATED AT 250 WORDS)

Relationships of plasma and hepatic variables with rates of plasma low-density lipoprotein apolipoprotein B metabolism in baboons fed low- and high-fat diets.

These studies were conducted to determine relationships of plasma low-density lipoprotein (LDL) cholesterol concentrations and hepatic mRNA levels for apolipoprotein (apo) B, LDL receptor, and hepatic hydroxymethyl glutaryl coenzyme A (HMG CoA) synthase with plasma LDL apo B production and catabolic rates in baboons maintained on a low-cholesterol, low-fat chow diet and on a high-cholesterol, high-fat (HCHF) diet. Twelve baboons with LDL cholesterol levels ranging from low to high on the HCHF diet but with similar high-density lipoprotein (HDL) cholesterol levels were selected from a colony of selectively bred pedigreed baboons. LDL apo B turnover and hepatic mRNA concentrations for apo B, LDL receptor, and HMG CoA synthase were measured on a chow diet and again on a HCHF diet fed for 14 weeks. LDL apo B fractional catabolic rates decreased and production rates increased on the HCHF diet. Hepatic mRNA concentrations for apo B were not affected by the HCHF diet. Hepatic LDL receptor and HMG CoA synthase mRNA concentrations decreased on the HCHF diet as compared with the chow diet. LDL apo B fractional catabolic rate was negatively correlated with plasma cholesterol, LDL cholesterol, LDL apo B, and LDL apo B production and positively correlated with hepatic LDL receptor and HMG CoA synthase mRNA concentrations and with plasma LDL triglyceride to cholesterol ratio on the chow diet but not on the HCHF diet. LDL apo B production was positively correlated with plasma cholesterol, LDL cholesterol, and LDL apo B on the HCHF diet and negatively correlated with LDL triglyceride to cholesterol ratio on both chow and HCHF diets.(ABSTRACT TRUNCATED AT 250 WORDS)

Total body water-to-lean body mass ratio in baboons (Papio sp.) of varying adiposity.

Total body water (TBW), lean body mass (LBM), and triglyceride mass were measured in 23 5-yr-old baboons (13 females and 10 males). Male baboons weighed more, had more LBM, more TBW, and contained less triglyceride mass per unit body weight than female baboons. Among all baboons, triglyceride mass per unit body weight ranged from 2.4 to 33.5%. The ratio TBW:LBM ranged from 0.70 to 0.92, increasing (r = 0.98) with increased body triglyceride content (both percent and absolute mass) in both male and female baboons. However, the water content per unit weight of tissues free of fat cells (liver, lungs, kidneys, central nervous system, eyes, tongue) was nearly constant at 0.73 +/- 0.02. The increase in TBW:LBM is more than can be expected from the water in the increased adipose tissue mass. We conclude that TBW:LBM is not constant but is influenced by body triglyceride content in baboons.

Use of NMR for measurement of total body water and estimation of body fat.

A nuclear magnetic resonance (NMR) method is described for quantitatively measuring total body water (TBW) and for estimating the fat content of baboons. The hydrogen associated with water was measured as the amplitude of the free-induction decay voltage following a series of 90 degree radio frequency pulses at the Lamour frequency for hydrogen with a pulse length of 14 microseconds and a peak measuring time of 50 microseconds. TBW was calculated by multiplying the peak amplitude (volts) by the experimentally determined constant for a water standard (g water/V). This NMR method yielded TBW contents similar to those obtained in the same baboons by direct gravimetric procedures. In contrast, the widely used 3H2O-dilution method usually and variably overestimated body water. By providing an accurate measure of body water, this NMR procedure provides a rapid, noninvasive, reasonably accurate way of estimating body fat content.

Resveratrol promotes atherosclerosis in hypercholesterolemic rabbits.

The hypothesis was tested that resveratrol, a compound in red wine, would inhibit atherosclerotic development in rabbits fed 0.5% cholesterol for 60 days. Rabbits were supplemented with or without oral resveratrol. During the study, body weights and food consumption were similar for the two groups. The lack of differences between liver weights and a series of serum parameters indicative of liver disease suggest that liver function was similar in the two groups. The diet produced hypercholesterolemia in both groups, but no differences in lipoprotein-cholesterol concentrations. The electrophoretic mobility of plasma low-density lipoprotein (LDL) and plasma LDL after induced oxidation also was not different between the groups. Staining of atherosclerotic lesions in the control and resveratrol-treated groups revealed that the resveratrol-treated rabbits had significantly more aortic surface area covered by atherosclerotic lesions (P < 0.02). Therefore, resveratrol promoted atherosclerotic development, rather than protect against it, by a mechanism that is independent of observed differences in gross animal health, liver function, plasma cholesterol concentrations, or LDL oxidative status.

Comparison of laboratory performance with blind and mail-distributed proficiency testing samples.

Simulated addict urine samples containing drugs were sent to collaborating hospital administrators and officials of methadone centers, who then forwarded the samples to their supporting laboratories as though they were ordinary specimens from patients. The laboratories, which were already participating in the proficiency testing program of the Center for Disease Control, received the identical test samples in the mail as part of a regular Center for Disease Control proficiency testing program. Most of the laboratories performed acceptably with the mail-distributed samples, but many performed poorly when the identical samples were sent to them as if they were specimens from patients. Because of the limitations of proficiency testing involving mail-distribution samples and the impracticality of extensive testing with blind samples on a national level, the Center for Disease Control proposes to compliment its regular proficiency testing program with a monitored, onsite program of performance evaluation.

Developmental changes in cholesterol 7alpha- and 27-hydroxylases in the piglet.

Hepatic cholesterol 7alpha-hydroxylase (CYP7A) and sterol 27 hydroxylase activities were measured in fetal, newborn, suckling, and weaned piglets from 76 d into gestation to 49 d of age. Hepatic CYP7A activity was not detected in fetal microsomes, but it increased to 6.8 +/- 2.6 pmol/min x mg(-1) protein in suckling piglets at 21 d of age and to 18.2 +/- 2.5 in weaned piglets at 49 d of age. Hepatic CYP7A activity was not different between 49-d-old piglets weaned at 21 d and piglets suckled for 49 d (18.9 +/- 2.6 and 18.2 +/- 2.5 pmol/min x mg protein, respectively). Fasting for 14 h decreased CYP7A activity by 86% in both suckled and weaned piglets. Cholesterol 7alpha-hydroxylase activity remained decreased for at least 5 h after refeeding. Sterol 27-hydroxylase activity was also undetectable near birth, but was detectable by 21 d of age. Postnatally, sterol 27-hydroxylase activity was not influenced by age or suckling and weaning, as was CYP7A. Sterol 27-hydroxylase was decreased by 80% in piglets deprived of feed compared with piglets given free access. In contrast to CYP7A activity, 27-hydroxylase activity returned within 5 h after refeeding to levels observed in piglets given ad libitum access to feed. Similar to CYP7A enzyme activity, hepatic CYP7A mRNA was not detected in newborn piglets, but increased from 2.7 +/- 1.7 pg mRNA/microg RNA in suckling piglets at 21 d to 13.7 +/- 1.2 in 49-d-old piglets weaned at 21 d. As with enzyme activity, feed deprivation decreased CYP7A mRNA to barely detectable levels (< .5 pg/microg RNA), and which remained decreased for at least 5 h following refeeding (.6 +/- .3 and 2.67 +/- .4 pg mRNA/microg RNA for suckled and weaned piglets, respectively). In piglets allowed free access to feed, CYP7A mRNA concentrations were associated positively (P = .001) with enzyme activity. These results suggest that developmental regulation of CYP7A activity is the result of a pretranslational mechanism.