ASAS centennial paper: animal growth and development research: historical perspectives.

From a historical perspective, it is difficult to identify a specific date that launched the field of endocrinology. One "biomarker" of the inception of endocrinology traces back to Ernest Henry Starling, who first introduced the word hormone in a talk given in 1905 at the Royal College of Physicians in London (Starling, 1905). A historical look at the field of endocrine regulation of animal growth since 1905 conveys that countless scientists worldwide worked to advance the scientific evidence base, which led to the commercial development of hormone-based products that enhanced growth and beneficially changed carcass composition of meat animals. This review will discuss some of seminal contributions that include the discovery of hormones (like ST and beta-adrenergic agonists) that have been shown to play key roles in regulating growth and nutrient partitioning of livestock, the mechanisms by which these hormones act, and the development of products for application in animal agriculture.

Somatotropic function: the somatomedin hypothesis revisited.

The discovery in 1922 that an alkaline extract of the anterior pituitary can increase growth and change carcass composition of rats led to the discovery of growth hormone (somatotropin, ST). Since the early studies, much has been learned about the biological effects of ST. The advent of recombinant DNA technology has led to the commercial development of ST-based products for animal agriculture. Administration of porcine ST (pST) at maximally effective doses (approximately 100 microg x kg BW(-1) x d(-1)) to growing pigs for 30 to 77 d increases average daily gain approximately 10 to 20%, improves productive efficiency (i.e., the ratio of body weight gain to feed consumed) 13 to 33%, decreases lipid accretion rates by as much as approximately 80%, and stimulates protein deposition (muscle growth) by as much as 70%. These responses are associated with a decrease in feed intake of approximately 10 to 15%. The effects of ST are mediated directly and indirectly. The indirect effects of ST are mediated by the somatomedin (insulin-like growth factor-I). The discovery of somatomedin led to the introduction of the somatomedin hypothesis, which explained the basis of ST action. Since the discovery of the somatomedins, there have been several modifications of the hypothesis developed to accommodate the evolution in understanding of how ST and IGF-I affect a diverse array of biological events. This review will summarize the history of ST and the evolution of the somatomedin hypothesis.

Bioactive compounds in nutrition and health-research methodologies for establishing biological function: the antioxidant and anti-inflammatory effects of flavonoids on atherosclerosis.

Identifying bioactive compounds and establishing their health effects are active areas of scientific inquiry. There are exciting prospects that select bioactive compounds will reduce the risk of many diseases, including chronic diseases such as cardiovascular disease. Recent findings have established that cardiovascular disease is a disease of inflammation, and consequently is amenable to intervention via molecules that have anti-inflammatory effects. In addition, research demonstrating adverse effects of oxidants on atherogenesis raises the possibility that antioxidants can confer cardioprotective effects. This review provides an overview of research approaches that can be used to unravel the biology and health effects of bioactive compounds. Because of the number of bioactive compounds and the diversity of likely biological effects, numerous and diverse experimental approaches must be taken to increase our understanding of the biology of bioactive compounds. Recognizing the complexity of this biology, sophisticated experimental designs and analytical methodologies must be employed to advance the field. The discovery of novel health effects of bioactive compounds will provide the scientific basis for future efforts to use biotechnology to modify/fortify foods and food components as a means to improve public health.

Dietary fat: assessing the evidence in support of a moderate-fat diet; the benchmark based on lipoprotein metabolism.

There is a growing database that has evaluated the effects of varying amounts of total fat on risk factors for cardiovascular disease, diabetes and overweight and obesity. The evidence clearly suggests that extremes in dietary fat should be avoided, and instead a diet moderate in total fat (25-35 % energy) is preferable for the majority of individuals. Moreover, we now appreciate the importance of individualizing dietary fat recommendations within this range of total fat. With respect to cardiovascular disease, a diet higher in total fat (30-35 % energy) affects the lipid and lipoprotein risk profile more favourably than a lower-fat diet; this is also the case for individuals with diabetes, with the added benefit of better glycaemic control. Dietary fibre (> or = 25 g/d) attenuates and even prevents the potentially adverse lipid and lipoprotein effects of a lower-fat diet. With respect to weight control, a moderate-fat diet can be as, or even more, effective than a lower-fat diet, because of advantages with long-term adherence and potentially favourable effects on lipids and lipoproteins. Thus, there is now a convincing scientific basis to advocate a diet moderate in total fat for the majority of individuals. Implicit to this position is that unsaturated fat has numerous beneficial health effects. However, because fat is energy dense, moderation in fat intake is essential for weight control. Consequently, a simple message to convey is to avoid diets that are very low and very high in fat. Moreover, within the range of a moderate-fat diet it is still important to individualize the total fat prescription. Nonetheless, the guiding principle is that moderation in total fat is the defining benchmark for a contemporary diet that reduces risk of chronic disease.

Porcine growth hormone: a central metabolic hormone involved in the regulation of adipose tissue growth.

During the past 20 y, much has been learned about how porcine growth hormone (pGH) affects growth and nutrient partitioning in growing pigs. Our contemporary understanding of the biology of pGH has as its roots the seminal studies conducted by Larry Machlin. His studies and many subsequent reports by other investigators have established that treatment of growing pigs with pGH markedly stimulates muscle growth and, concurrently, reduces fat deposition. In growing pigs, maximally effective doses of pGH increase average daily gain as much as 10% to 20%, improve feed efficiency 15% to 30%, decrease adipose tissue mass and lipid accretion rates by as much as 50% to 80%, and concurrently increase protein deposition by 50%. These effects are associated with a decrease in feed intake of approximately 10% to 15%. These responses occur because pGH has a wide array of biological effects that modulate nutrient partitioning between adipose tissue and skeletal muscle. The decrease in adipose tissue growth is due to a reduction in lipogenesis that is the consequence of pGH blunting the effects of many insulin-dependent events. This article provides an overview of some of the biological effects pGH has in adipose tissue and discusses what is known about the underlying mechanisms that account for these effects.

Effects of cocoa powder and dark chocolate on LDL oxidative susceptibility and prostaglandin concentrations in humans.

BACKGROUND: Flavonoids are polyphenolic compounds of plant origin with antioxidant effects. Flavonoids inhibit LDL oxidation and reduce thrombotic tendency in vitro. Little is known about how cocoa powder and dark chocolate, rich sources of polyphenols, affect these cardiovascular disease risk factors. OBJECTIVE: We evaluated the effects of a diet high in cocoa powder and dark chocolate (CP-DC diet) on LDL oxidative susceptibility, serum total antioxidant capacity, and urinary prostaglandin concentrations. DESIGN: We conducted a randomized, 2-period, crossover study in 23 healthy subjects fed 2 diets: an average American diet (AAD) controlled for fiber, caffeine, and theobromine and an AAD supplemented with 22 g cocoa powder and 16 g dark chocolate (CP-DC diet), providing approximately 466 mg procyanidins/d. RESULTS: LDL oxidation lag time was approximately 8% greater (P = 0.01) after the CP-DC diet than after the AAD. Serum total antioxidant capacity measured by oxygen radical absorbance capacity was approximately 4% greater (P = 0.04) after the CP-DC diet than after the AAD and was positively correlated with LDL oxidation lag time (r = 0.32, P = 0.03). HDL cholesterol was 4% greater after the CP-DC diet (P = 0.02) than after the AAD; however, LDL-HDL ratios were not significantly different. Twenty-four-hour urinary excretion of thromboxane B(2) and 6-keto-prostaglandin F(1)(alpha) and the ratio of the 2 compounds were not significantly different between the 2 diets. CONCLUSION: Cocoa powder and dark chocolate may favorably affect cardiovascular disease risk status by modestly reducing LDL oxidation susceptibility, increasing serum total antioxidant capacity and HDL-cholesterol concentrations, and not adversely affecting prostaglandins.

Transcriptional regulation of fatty acid synthase gene by somatotropin in 3T3-F442A adipocytes.

Somatotropin (ST) antagonizes insulin stimulation of fatty acid synthase (FAS) enzyme activity and gene transcription in adipocytes. Previous studies have shown that an insulin response element (IRE) is located in the proximal region of the FAS promoter (-71 to -50) and upstream stimulatory factor (USF) 1 binds to this IRE. The present study was conducted to initially evaluate whether there is a somatotropin response element (STRE) in the 5'flanking region of the FAS gene and to determine whether USF1 mediates the effect of ST on FAS gene transcription in 3T3-F442A adipocytes. Two 5' deletion FAS promoter constructs (pFAS-CATS4 and pFAS-CAT5), which contain the 5' flanking sequences of the rat FAS gene at -112 to +65 and -2195 to +65, respectively, were stably transfected into 3T3-F442A preadipocytes. Insulin stimulated chloramphenicol acetyltransferase (CAT) activity 1.7- and 4.7-fold (P < 0.05) in 3T3-F442A adipocytes transfected with pFAS-CATS4 and pFAS-CAT5, respectively. In contrast, bovine somatotropin (bST) attenuated the stimulatory effect of insulin on CAT activity by approximately 60% (P < 0.05) in both constructs. When 3T3-F442A adipocytes were treated with insulin (10 ng/mL) or insulin (10 ng/mL) plus bST (100 ng/mL) for 24, 48, or 72 h, neither insulin nor bST significantly affected USF1 mRNA levels. When human USF1 (hUSF1) cDNA probe was used, however, insulin increased the abundance of an unidentified transcript (named hUSF1-like mRNA) 11- to 25-fold (P < 0.05) and ST decreased the stimulatory effect of insulin on hUSF1-like mRNA levels by 50 to 90% (P < 0.05). Western blot analyses of nuclear extracts from cells treated with insulin (10 ng/mL) or insulin (10 ng/mL) plus bST (100 ng/mL) for 48 h demonstrated that the abundance of USF1 was not affected by insulin or ST. Furthermore, electrophoretic mobility shift analyses (EMSA) of nuclear extracts revealed that neither insulin nor ST had an effect on the binding of USF1 to the IRE. These results suggest that a STRE may be located within the first 112 bp of the FAS promoter and that USF1 does not directly mediate the effect of ST on transcription of the FAS gene in 3T3-F442A adipocytes.

Low fat and high monounsaturated fat diets decrease human low density lipoprotein oxidative susceptibility in vitro.

Oxidative modification of low density lipoprotein (LDL) is thought to play an important role in the development of atherosclerosis. Some studies have found that LDL enriched in monounsaturated fatty acids (MUFA) are less susceptible to oxidation than LDL enriched in polyunsaturated fatty acids (PUFA). A high MUFA diet is an alternative to a lower-fat blood cholesterol-lowering diet. Less is known about the effects of high MUFA versus lower-fat blood cholesterol-lowering diets on LDL oxidative susceptibility. The present study was designed to evaluate the effects of men and women consuming diets high in MUFA (peanuts plus peanut butter, peanut oil and olive oil) on LDL oxidative susceptibility, and to compare these effects with those of a Step II blood cholesterol-lowering diet. A randomized, double-blind, five-period crossover design (n = 20) was used to study the effects of the following diets on LDL-oxidation: average American [35% fat, 15% saturated fatty acids (SFA)], Step II (25% fat, 7% SFA), olive oil (35% fat, 7% SFA), peanut oil (35% fat, 7% SFA) and peanuts plus peanut butter (35% fat, 8% SFA). The average American diet resulted in the shortest lag time (57 +/- 6 min) for LDL oxidized ex vivo, whereas the Step II, olive oil and peanuts plus peanut butter diets resulted in a lag time of 66 +/- 6 min (P < or = 0.1). The slower rate of oxidation [nmol dienes/(min x mg LDL protein)] observed when subjects consumed the olive oil diet (24 +/- 2) versus the average American (28 +/- 2), peanut oil (28 +/- 2) and peanuts plus peanut butter diets (29 +/- 2; P < or = 0.05) was associated with a lower LDL PUFA content. The results of this study suggest that lower-fat and higher-fat blood cholesterol-lowering diets high in MUFA have similar effects on LDL oxidative resistance. In addition, our results suggest that different high MUFA sources varying in the ratio of MUFA to PUFA can be incorporated into a high MUFA diet without increasing the susceptibility of LDL to oxidation.

The effects of nuts on coronary heart disease risk.

Epidemiologic studies have consistently demonstrated beneficial effects of nut consumption on coronary heart disease (CHD) morbidity and mortality in different population groups. Clinical studies have reported total and low-density lipoprotein cholesterol-lowering effects of heart-healthy diets that contain various nuts or legume peanuts. It is evident that the favorable fatty acid profile of nuts (high in unsaturated fatty acids and low in saturated fatty acids) contributes to cholesterol lowering and, hence, CHD risk reduction. Dietary fiber and other bioactive constituents in nuts may confer additional cardioprotective effects.

Analysis of the signal pathways involved in the regulation of fatty acid synthase gene expression by insulin and somatotropin.

Our previous studies have shown that somatotropin (ST) antagonizes insulin stimulation of fatty acid synthase (FAS) enzyme activity and gene transcription in adipocytes. In the present study, inhibitors of insulin and ST signaling pathways were used to dissect the mechanisms by which these hormones regulate FAS gene expression in 3T3-F442A adipocytes. Treating 3T3-F442A adipocytes with 10 microM PD98059, an inhibitor of mitogen-activated protein (MAP) kinase, did not affect the induction of FAS mRNA by insulin. When cells were cultured with H-89 (10 microM), GF109203X (10 microM), or staurosporine (100 nM), inhibitors of protein kinase A, protein kinase C, and Janus kinase (JAK) 2, respectively, the inhibitory effect of ST on FAS mRNA levels was not altered. However, H-89 significantly decreased the stimulatory effect of insulin on FAS mRNA abundance. Moreover, treatment with okadaic acid (1 microM), a serine/threonine phosphatase inhibitor, abolished the induction of FAS mRNA by insulin. These results suggest that serine/threonine dephosphorylation and protein kinase A-dependent pathways are involved in the regulation of FAS gene expression by insulin, but MAP kinase is probably not involved. Furthermore, our data indicate that protein kinase A, protein kinase C, and JAK2 do not mediate the effect of ST on regulation of FAS mRNA abundance.

The biology of somatotropin in adipose tissue growth and nutrient partitioning.

During the past 20 years, much has been learned about how porcine somatotropin (pST) affects growth and nutrient partitioning in growing pigs. The development of techniques to produce large quantities of recombinantly derived pST enabled numerous long-term studies to be conducted in which the effects of daily pST administration could be evaluated. Collectively, these studies established that treatment of growing pigs with pST markedly stimulated muscle growth and, concurrently, reduced fat deposition. In growing pigs, maximally effective doses of pST increase average daily gain as much as 10-20%, improve feed efficiency 15-30%, decrease adipose tissue mass and lipid accretion rates by as much as 50-80% and concurrently increase protein deposition by 50%. These effects are associated with a decrease in feed intake of approximately 10-15%. These responses occur because pST has a wide array of biological effects that modulate nutrient partitioning between adipose tissue and skeletal muscle. The decrease in adipose tissue growth is due to a reduction in lipogenesis that is the consequence of pST blunting the effects of many insulin-dependent events. With respect to fatty acid synthase (FAS), a pace-setting enzyme in the lipogenic pathway, enzyme activity is markedly reduced by pST. This is the result of a pST-mediated decrease in FAS mRNA levels that occurs because FAS gene transcription is decreased. The consequence of the decrease in lipid synthesis is that adipocyte hypertrophy is impaired and, hence, tissue growth. This review will provide an overview of some of the biological effects of pST in adipose tissue and will discuss what is known about the underlying mechanisms that account for these effects.

Lowering dietary saturated fat and total fat reduces the oxidative susceptibility of LDL in healthy men and women.

The present study examined the effects of reducing dietary total fat and saturated fat (SFA) on LDL oxidative susceptibility in 27 healthy men and women (age 24-65 y). Each subject consumed each of three diets for 8 wk: an average American diet (AAD, 34% energy from fat, 15% from SFA), a Step-1 diet (29% fat, 9% SFA) and a very low SFA diet (Low-Sat, 25% fat, 6% SFA). In vitro LDL oxidation was assessed by copper-mediated oxidation, as measured by the kinetics of conjugated diene formation and lipid peroxide formation. Compared with the AAD, plasma LDL-cholesterol (LDL-C) and HDL cholesterol levels were 8% lower (P: = 0.16 and P: = 0.11, respectively), in subjects when they consumed the Step-1 diet and 11% (P: < 0.03) and 14% (P: < 0.057) lower, respectively, when they consumed the Low-Sat diet. Conjugated diene production and oxidation rate were 7% (P: < 0. 05) and 9% (P: < 0.05) lower, respectively. The reduction of lipid peroxide formation was 9% (P: < 0.05) in subjects when they consumed the Low-Sat diet vs. the AAD. In addition, lipid peroxide and conjugated diene formation were positively correlated with plasma total and LDL-C and apolipoprotein B (apo B) levels (r = 0.5-0.6, P: < 0.001), suggesting that quantity of LDL is an important determinant of oxidative modification. Furthermore, at the same level of apo B or LDL-C, LDL from subjects when they consumed either Step-1 or Low-Sat diets was less susceptible (P: < 0.05) to oxidation than those when they consumed the AAD, suggesting that qualitative changes also affect LDL oxidative susceptibility. Therefore, the benefits of lowering dietary SFA may extend beyond decreasing LDL-C levels and include favorable qualitative changes in LDL that further decrease risk of coronary heart disease.

Polyunsaturated fatty acids in the food chain in the United States.

In the United States, intake of n-3 fatty acids is approximately 1.6 g/d ( approximately 0.7% of energy), of which 1.4 g is alpha-linolenic acid (ALA; 18:3) and 0.1-0.2 g is eicosapentaenoic acid (EPA; 20:5) and docosahexaenoic acid (DHA; 22:6). The primary sources of ALA are vegetable oils, principally soybean and canola. The predominant sources of EPA and DHA are fish and fish oils. Intake data indicate that the ratio of n-6 to n-3 fatty acids is approximately 9.8:1. Food disappearance data between 1985 and 1994 indicate that the ratio of n-6 to n-3 fatty acids has decreased from 12.4:1 to 10.6:1. This reflects a change in the profile of vegetable oils consumed and, in particular, an approximate 5.5-fold increase in canola oil use. The ratio of n-6 to n-3 fatty acids is still much higher than that recommended (ie, 2.3:1). Lower ratios increase endogenous conversion of ALA to EPA and DHA. Attaining the proposed recommended combined EPA and DHA intake of 0.65 g/d will require an approximately 4-fold increase in fish consumption in the United States. Alternative strategies, such as food enrichment and the use of biotechnology to manipulate the EPA and DHA as well as ALA contents of the food supply, will become increasingly important in increasing n-3 fatty acid intake in the US population.

High-monounsaturated fatty acid diets lower both plasma cholesterol and triacylglycerol concentrations.

BACKGROUND: Low-fat diets increase plasma triacylglycerol and decrease HDL-cholesterol concentrations, thereby potentially adversely affecting cardiovascular disease (CVD) risk. High-monounsaturated fatty acid (MUFA), cholesterol-lowering diets do not raise triacylglycerol or lower HDL cholesterol, but little is known about how peanut products, a rich source of MUFAs, affect CVD risk. OBJECTIVE: The present study compared the CVD risk profile of an Average American diet (AAD) with those of 4 cholesterol-lowering diets: an American Heart Association/National Cholesterol Education Program Step II diet and 3 high-MUFA diets [olive oil (OO), peanut oil (PO), and peanuts and peanut butter (PPB)]. DESIGN: A randomized, double-blind, 5-period crossover study design (n = 22) was used to examine the effects of the diets on serum lipids and lipoproteins: AAD [34% fat; 16% saturated fatty acids (SFAs), 11% MUFAs], Step II (25% fat; 7% SFAs, 12% MUFAs), OO (34% fat; 7% SFAs, 21% MUFAs), PO (34% fat; 7% SFAs, 17% MUFAs), and PPB (36% fat; 8% SFAs, 18% MUFAs). RESULTS: The high-MUFA diets lowered total cholesterol by 10% and LDL cholesterol by 14%. This response was comparable with that observed for the Step II diet. Triacylglycerol concentrations were 13% lower in subjects consuming the high-MUFA diets and were 11% higher with the Step II diet than with the AAD. The high-MUFA diets did not lower HDL cholesterol whereas the Step II diet lowered it by 4% compared with the AAD. The OO, PO, and PPB diets decreased CVD risk by an estimated 25%, 16%, and 21%, respectively, whereas the Step II diet lowered CVD risk by 12%. CONCLUSION: A high-MUFA, cholesterol-lowering diet may be preferable to a low-fat diet because of more favorable effects on the CVD risk profile.

Emerging strategies for enhancing growth: is there a biotechnology better than somatotropin?

During the past 20 years, there have been many impressive advances in a number of scientific disciplines that have led to the discovery and development of exciting new biotechnologies that offer the potential to improve productive efficiency of animal agriculture. Some technologies have been developed from advances made in our understanding of how the endocrine system regulates growth and lactation. This information then has been used to devise viable strategies that alter circulating hormone concentration(s) to enhance animal production and productive efficiency. The most notable success to date using this approach has been bovine somatotropin, which has been adopted for use in the dairy industry in certain countries. Advances in transgenic biology, gene therapy, "knock-out" gene technologies, and cloning may lead to other novel products/strategies that enhance productive efficiency. The purpose of this paper is to discuss what future strategies might emerge that will increase meat and milk production and the efficiency of these processes.

Nuts and their bioactive constituents: effects on serum lipids and other factors that affect disease risk.

Because nuts have favorable fatty acid and nutrient profiles, there is growing interest in evaluating their role in a heart-healthy diet. Nuts are low in saturated fatty acids and high in monounsaturated and polyunsaturated fatty acids. In addition, emerging evidence indicates that there are other bioactive molecules in nuts that elicit cardioprotective effects. These include plant protein, dietary fiber, micronutrients such as copper and magnesium, plant sterols, and phytochemicals. Few feeding studies have been conducted that have incorporated different nuts into the test diets to determine the effects on plasma lipids and lipoproteins. The total- and lipoprotein-cholesterol responses to these diets are summarized in this article. In addition, the actual cholesterol response was compared with the predicted response derived from the most current predictive equations for blood cholesterol. Results from this comparison showed that when subjects consumed test diets including nuts, there was an approximately 25% greater cholesterol-lowering response than that predicted by the equations. These results suggest that there are non-fatty acid constituents in nuts that have additional cholesterol-lowering effects. Further studies are needed to identify these constituents and establish their relative cholesterol-lowering potency.

Biology of somatotropin in growth and lactation of domestic animals.

Impressive progress has been made during the past 15 years in our understanding of the biology of somatotropin (ST) in domestic animals. In part, this progress was sparked by advances in biotechnology that made feasible the production of large quantities of recombinant bovine ST (bST) and porcine ST (pST). The availability of recombinant bST and pST resulted in an exponential increase in investigations that explored their role in growth and lactation biology, as well as evaluated their potential for commercial use. Collectively, these studies established that administration of bST to lactating dairy cows increased milk yield, and treatment of growing pigs with pST markedly stimulated muscle growth and reduced fat deposition. In addition to these "efficacy" studies, a substantial number of investigations examined the mechanisms by which ST affects lactation and growth of domestic animals. This review summarizes the diverse physiological effects ST has on growth and lactation and discusses the underlying mechanisms that mediate these effects in domestic animals.

Somatotropin-dependent decrease in fatty acid synthase mRNA abundance in 3T3-F442A adipocytes is the result of a decrease in both gene transcription and mRNA stability.

Somatotropin (ST) markedly decreases lipogenesis, fatty acid synthase (FAS) enzyme activity and mRNA abundance in pig adipocytes. The present study was conducted to determine whether the decrease in FAS mRNA in 3T3-F442A adipocytes was the result of a decrease in transcription of the FAS gene and/or a change in FAS mRNA stability. Insulin increased the abundance of FAS mRNA 2-13-fold and fatty acid synthesis 3-7-fold. Somatotropin decreased the stimulatory effect of insulin on the abundance of FAS mRNA and lipogenesis by 40-70% and 20-60% respectively. Subsequent run-on analyses demonstrated that the decrease observed in FAS mRNA in response to ST was associated with an 82% decrease in transcription; ST significantly shortened the half-life of FAS mRNA from 35 to 11 h. To corroborate the run-on analyses, cells were stably transfected with a pFAS-CAT5 (in which CAT stands for chloramphenicol acetyltransferase) reporter construct that contained 2195 bp of the 5' flanking region of the rat FAS gene. Insulin treatment increased FAS-CAT activity 4.7-fold. When ST was added to the insulin-containing medium there was an approx. 60% reduction in FAS-CAT activity. In summary, our results indicate that ST decreases FAS mRNA levels and that this is the result of a marked decrease in both transcription of the FAS gene and stability of the FAS mRNA.