Diet-induced alterations in progesterone clearance appear to be mediated by insulin signaling in hepatocytes

Factors that affect progesterone clearance from plasma and by hepatocytes in culture were examined in a series of experiments. In Exp. 1, the objective was to determine whether an increase in hepatic portal blood acetate or propionate could alter progesterone metabolism by the liver. For ewe lambs gavaged orally with sodium propionate compared with those gavaged orally with sodium acetate, serum progesterone concentrations began to diverge as early as 0.5 h after administration and were greater (P < 0.05) at 3 and 4 h after administration. The objective of Exp. 2 was to determine the effect of a single oral gavage of either sodium acetate or sodium propionate on peripheral insulin and glucagon concentrations. Ewes gavaged orally with sodium propionate had greater (P < 0.05) insulin concentrations at 0.5 and 1 h after gavage than ewes gavaged with sodium acetate. Furthermore, glucagon concentrations were greater (P < 0.05) at 0.5, 1, and 2 h for ewe lambs gavaged orally with sodium propionate compared with those receiving sodium acetate. The third experiment investigated the rate of in vitro progesterone clearance by cultured hepatocytes in response to treatment with different concentrations of insulin and glucagon. Progesterone clearance was reduced (P < 0.05) with the addition of 0.1 nM insulin compared with the control. Furthermore, there was a greater reduction (P < 0.05) in progesterone clearance in response to 1.0 and 10 nM insulin compared with the control and 0.1 nM insulin. No change was observed in progesterone clearance in hepatocytes treated with either physiological (0.01 and 0.1 nM) or supraphysiological (1.0 nM) glucagon. Supraphysiological concentrations of glucagon (1.0 nM) negated the effects of either 0.1 or 1.0 nM insulin on progesterone clearance by hepatocytes. However, with physiological concentrations of glucagon (0.1 nM) and 1.0 nM insulin, glucagon was not able to negate the reduction in progesterone clearance caused by insulin. These data are consistent with a paradigm in which elevated hepatic portal vein propionate increases plasma insulin in ruminants, which decreases progesterone clearance, thereby increasing serum progesterone concentrations.     

Comparative lipid and lipoprotein metabolism

This review summarizes a number of recent reports in several areas of lipid and lipoprotein metabolism. Absorption of dietary lipids, cholesterol synthesis, and biliary cholesterol metabolism are mentioned only briefly to be complete. Comparative aspects of lipoprotein metabolism, however, are detailed in an effort to integrate the myriad metabolic events which characterize these important lipid transport particles. Where comparative information is known, those aspects of lipoprotein metabolism that may be protective against atherogenesis in certain mammalian species are also described. Efforts to understand atherogenic resistance comparatively in animals lends a better understanding of the metabolic events leading to coronary artery disease in humans. They also provide an important basis for understanding lipid metabolism in numerous veterinary species.     

Diet-induced bacterial immunogens in the gastrointestinal tract of dairy cows: Impacts on immunity and metabolism

Dairy cows are often fed high grain diets to meet the energy demand for high milk production or simply due to a lack of forages at times. As a result, ruminal acidosis, especially subacute ruminal acidosis (SARA), occurs frequently in practical dairy production. When SARA occurs, bacterial endotoxin (or lipopolysaccharide, LPS) is released in the rumen and the large intestine in a large amount. Many other bacterial immunogens may also be released in the digestive tract following feeding dairy cows diets containing high proportions of grain. LPS can be translocated into the bloodstream across the epithelium of the digestive tract, especially the lower tract, due to possible alterations of permeability and injuries of the epithelial tissue. As a result, the concentration of blood LPS increases. Immune responses are subsequently caused by circulating LPS, and the systemic effects include increases in concentrations of neutrophils and the acute phase proteins such as serum amyloid-A (SAA), haptoglobin (Hp), LPS binding protein (LBP), and C-reactive protein (CRP) in blood. Entry of LPS into blood can also result in metabolic alterations. Blood glucose and nonesterified fatty acid concentrations are enhanced accompanying an increase of blood LPS after increasing the amount of grain in the diet, which adversely affects feed intake of dairy cows. As the proportions of grain in the diet increase, patterns of plasma β-hydoxybutyric acid, cholesterol, and minerals (Ca, Fe, and Zn) are also perturbed. The bacterial immunogens can also lead to reduced supply of nutrients for synthesis of milk components and depressed functions of the epithelial cells in the mammary gland. The immune responses and metabolic alterations caused by circulating bacterial immunogens will exert an effect on milk production. It has been demonstrated that increases in concentrations of ruminal LPS and plasma acute phase proteins (CRP, SAA, and LBP) are associated with declines in milk fat content, milk fat yield, 3.5% fat-corrected milk yield, as well as milk energy efficiency.     

A review of lipoprotein cholesterol metabolism: importance to ovarian function

Cholesterol utilized for steroid synthesis by ovarian tissue may be derived from de novo synthesis or cellular uptake of lipoprotein cholesterol. The majority of blood cholesterol is transported by either low (LDL) or high (HDL) density lipoproteins, depending on the animal species. Prior to vascularization, only HDL are in follicular fluid and contribute sterol to granulosa cells because other lipoproteins are unable to transverse the basement membrane due to their molecular masses. Following vascularization, both LDL and HDL bathe luteal cells. Most species preferentially use LDL cholesterol as a precursor for ovarian steroid synthesis. The LDL uptake by ovarian tissue occurs by receptor-mediated endocytosis. The receptor recognizes apolipoprotein B of LDL and apolipoprotein E found on some, but not all, HDL. Within a species, a positive relationship may exist between HDL apolipoprotein E content and importance of HDL cholesterol as a precursor for steroidogenesis. A "HDL pathway" exists for uptake of sterol from HDL void of apolipoprotein E. The HDL receptor exhibits broad binding specificity. Unlike LDL, the HDL particle is not internalized, and cholesterol preferentially is taken up relative to other HDL constituents. In most species, lipoproteins, rather than de novo synthesis from acetate, contribute the majority of cholesterol used for steroid production. Trophic hormones increase lipoprotein binding, internalization, degradation and conversion of lipoprotein-derived sterol to steroids, effects that are mediate through cyclic adenosine monophosphate. Knowledge recently acquired regarding lipoprotein sterol utilization by the ovary may be useful in developing nutritional, pharmacological or endocrine manipulations that may positively affect cholesterol clearance by the ovary, steroidogenesis and reproductive performance.     

Effect of hypothyroidism on kinetics of metabolism of very-low-density lipoprotein in mares

OBJECTIVE: To compare kinetics of the metabolism of very-low-density lipoprotein (VLDL) apolipoprotein B (apoB) before and after thyroidectomy in mares. ANIMALS: 5 healthy adult mares. PROCEDURE: Thyroidectomy was performed in euthyroid mares. Kinetics of VLDL apoB metabolism were measured before and after thyroidectomy by use of a bolus IV injection of 5,5,5-2H3 (98%) leucine (5 mg/kg) and subsequent isolation of labeled amino acid from plasma and VLDL. Labeled leucine was quantified by use of gas chromatography-mass spectrometry. Production rate (PR), delay time, and fractional catabolic rate (FCR) were calculated for the 2 forms of equine VLDL, apoB-48 VLDL, and apoB-100 VLDL. Plasma lipid concentrations were measured, and VLDL composition was determined. RESULTS: Physical appearance of horses was not altered by thyroidectomy. Significantly lower mean blood concentrations of thyroid hormones and non-esterified fatty acids were detected following thyroidectomy. Mean percentage of free cholesterol in VLDL was significantly higher after thyroidectomy. Mean plasma VLDL concentration or kinetics of apoB-48 or apoB-100 were not significantly altered by thyroidectomy. Mean +/- SEM PR was significantly lower (8.70 +/- 1.61 mg/kg/d) and mean delay time significantly longer (1.58 +/- 0.12 hours) for apoB-48 VLDL in euthyroid mares, compared with values for thyroidectomized mares (16.15 +/- 2.24 mg/kg/d and 0.93 +/- 0.10 hours, respectively). CONCLUSIONS AND CLINICAL RELEVANCE: Hypothyroidism did not significantly alter plasma VLDL concentrations or kinetics of VLDL apoB metabolism. Metabolism of apoB-48 VLDL differed significantly from that of apoB-100 VLDL in euthyroid mares.     

A Hepatocellular Adenoma in a Diet-induced Obese Mouse

A hepatic nodule was noted in a C57BL/6J mouse with diet-induced obesity at 53 weeks of age. Macroscopically, a protruding yellowish white nodule was observed on the visceral surface of the left lateral lobe. Light microscopy demonstrated clear demarcation from the compressed adjacent parenchyma, with loss of the distinct lobular pattern. The proliferating cells of the lesion varied in shape and showed cellular atypia and prominent nucleoli along with vacuoles of various sizes. Some of the cells contained various-sized eosinophilic inclusion bodies in their cytoplasm, and electron microscopy revealed the presence of lipid droplets in the rough endoplasmic reticulum. Eosinophilic inclusions were observed as electron dense granular material in the rough endoplasmic reticulum, with one or a few low density central cores. A diagnosis of hepatocellular adenoma was made based on these findings.     

Diet-induced central obesity and insulin resistance in rabbits

The present study was designed to examine whether rabbits fed a diet containing high fat and sucrose could develop obesity and insulin resistance (IR), the major pathophysiological features of metabolic syndrome. Male Japanese white rabbits were fed either a normal chow diet (control) or high fat and sucrose diet (HFSD) for 36 weeks. Plasma levels of triglycerides (TG), total cholesterol (TC), glucose and insulin were measured. To evaluate glucose metabolism, we performed an intravenous glucose tolerance test. In addition, we compared adipose tissue accumulation in HFSD-fed rabbits with that in normal rabbits. HFSD constantly and significantly led to an increase in body weight of HFSD-fed rabbits, caused by significantly higher visceral adipose tissue accumulation. Although there were no differences in plasma TG, TC, glucose, insulin levels and blood pressure between the two groups, HFSD-fed rabbits showed impaired glucose clearance associated with higher levels of insulin secretion compared to control rabbits. Our results showed that HFSD induced IR and increased adipose accumulation in rabbits, suggesting that HFSD-fed rabbits may become a model for research on human IR and obesity.     

Changes in lipoprotein metabolism and body composition in chickens in response to divergent selection for plasma very low density lipoprotein concentration

1. Plasma lipoprotein metabolism and body composition in lines of chicken selected for high- and low-plasma very low density lipoprotein (VLDL) concentrations were compared to the commercial broiler (meat-type) line from which they were derived. 2. Selection for low-plasma VLDL concentration for 10 generations has reduced the rate of VLDL secretion by at least 50% in males whereas selection for high-VLDL concentration has increased the rate of VLDL secretion over 2-fold. 3. Body fat content was highly correlated with rate of secretion of plasma triglyceride-rich (TGR) lipoproteins (r = 0.88 over the three lines). However, extrapolation of the data suggests that birds secreting no TGR-lipoproteins into the plasma would still have substantial amounts of body fat. 4. Selection for high VLDL has increased the proportion of circulating VLDL-triglyceride taken up by the abdominal fat pad by over 2-fold but there was no difference between high- and low-VLDL lines in the proportion of VLDL-triglyceride taken up by tissues and oxidised to [14C]-carbon dioxide. 5. The results confirm the importance of plasma lipoprotein metabolism in determining body composition in the chicken but suggest there are limits to further reduction in body fat content by manipulation of plasma lipoprotein metabolism.     

Impact of hyperlipidaemia on intermediary metabolism,faecal microbial metabolites and urinary characteristics of lipoprotein lipase deficient vs. normal cats

Findings in humans and rats indicate that hyperlipidaemia may be associated with enhanced endogenous oxalate (Ox) synthesis, which may be relevant for calcium oxalate (CaOx) urolith formation. Moreover, changes in lipid metabolism are proposed to negatively affect gut microbiota. This study aimed to investigate those potential interactions in hyperlipidaemic cats. Therefore, 10 normal control cats and seven lipoprotein lipase (LPL)‐deficient cats were fed a low‐fat diet for seven weeks. During the last week of the study, cats were housed in metabolic cages to collect urine and faeces. Blood was taken on the last day of the study. The LPL‐deficient cats had significantly higher serum triglyceride concentrations than normal cats, while lactate dehydrogenase (LDH) activity was not different. Urinary relative supersaturation with CaOx, urinary Ox, calcium, and citrate excretions, and urine pH did not differ between groups. Lower faecal acetic, propionic and total short‐chain fatty acid concentrations were observed in the LPL‐deficient cats. In conclusion, hyperlipidaemia does not appear to be a specific risk factor for CaOx urolith formation in cats. In contrast to results in rats, hyperlipidaemia was not accompanied by elevated serum LDH activity. As LDH can synthesise Ox from glycolate or other precursors, this might be one possible explanation for the similar urinary parameters in the LPL‐deficient and normal cats. Non‐diet‐induced hyperlipidaemia was not associated with marked changes in faecal microbial metabolites, suggesting no differences in the composition of the intestinal microbiota.     

Integrative analysis of indirect calorimetry and metabolomics profiling reveals alterations in energy metabolism between fed and fasted pigs

Background: Fasting is a simple metabolic strategy that is used to estimate the maintenance energy requirement where the energy supply for basic physiological functions is provided by the mobilization of body reserves.However, the underlying metabolic components of maintenance energy expenditure are not clear. This study investigated the differences in heat production(HP), respiratory quotient(RQ) and plasma metabolites in pigs in the fed and fasted state, using the techniques of indirect calorimetry and metabolomics.Methods: Nine barrows(45.2 ± 1.7 kg BW) were fed corn-soybean based meal diets and were kept in metabolism crates for a period of 14 d. After 7 d adaptation, pigs were transferred to respiratory chambers to determine HP and RQ based on indirect calorimetry. Pigs were fed the diet at 2,400 k J ME/(kg BW0.6·d) during d 8 to 12. The last 2 d were divided into 24 h fasting and 48 h fasting treatment, respectively. Plasma samples of each pig were collected from the anterior vena cava during the last 3 d(1 d while pigs were fed and 2 d during which they were fasted).The metabolites of plasma were determined by high-resolution mass spectrometry using a metabolomics approach.Results: Indirect calorimetry analysis revealed that HP and RQ were no significant difference between 24 h fasting and 48 h fasting, which were lower than those of fed state(P 0.01). The nitrogen concentration of urine tended to decrease with fasting(P = 0.054). Metabolomics analysis between the fed and fasted state revealed differences in15 compounds, most of which were not significantly different between 24 h fasting and 48 h fasting. Identified compounds were enriched in metabolic pathways related to linoleic acid metabolism, amino acid metabolism,sphingolipid metabolism, and pantothenate and Co A biosynthesis.Conclusion: These results suggest that the decreases in HP and RQ of growing pigs under fasting conditions were associated with the alterations of linoleic acid metabolism and amino acid metabolism. The integrative analysis also revealed that growing pigs under a 24-h fasting were more appropriate than a 48-h fasting to investigate the metabolic components of maintenance energy expenditure.     

Occurrence of retained placenta is preceded by an inflammatory state and alterations of energy metabolism in transition dairy cows

Background: Failure to expel fetal membranes within 24 h of calving is a pathological condition defined as retained placenta(RP). The objective of this investigation was to evaluate whether there are alterations in several selected serum variables related to innate immunity and carbohydrate and lipid metabolism that precede occurrence of RP in transition Holstein dairy cows.Methods: One hundred multiparous Holstein dairy cows were involved in the study. Blood samples were collected from the coccygeal vein during the-8 to +4 wks around parturition, once per week before the morning feeding.Six healthy control cows(CON) and 6 cows with RP were selected and serum samples at-8,-4, time of diagnosis of disease, and +4 wks relative to parturition were used for analyses. All samples were analyzed for lactate, nonesterified fatty acids(NEFA), β-hydroxybutyrate(BHBA), interleukin-1(IL-1), interleukin-6(IL-6), tumor necrosis factor(TNF), haptoglobin(Hp), and serum amyloid A(SAA).Results: Cows with RP had greater concentrations of serum lactate, IL-1, IL-6, TNF, and SAA in comparison with CON cows. Intriguingly, elevated concentrations of all five variables were observed at-8 and-4 wks before the occurrence of RP compared to healthy cows. Cows with RP also had lower DMI and milk production vs CON animals; however milk composition was not affected by RP.Conclusions: Cows with RP showed an activated innate immunity 8 wks prior to diagnosis of disease. Overall results suggest that serum IL-1, IL- 6, and TNF, and lactate can be used as screening biomarkers to indicate cows that might have health issues during the transition period.     

Congenital lipoprotein lipase deficiency in hyperlipemic kitten siblings.

The nature of the hyperlipemia in two 3-week-old male kittens, one of which was presented with the owner's complaints of retarded growth and white streaks on its eyes, was studied. Hypertriglyceridemia, hypercholesterolemia, and reduced Post-Heparin Plasma Lipolytic Activities (PHPLA) were observed in both animals. One of the kittens, however, was more severely affected and in addition, had lipemia retinalis and a marked lactescent, hyperchylomicronemic serum in spite of a short-term fast before sampling. These findings are strikingly similar to those found in human Type I hyperlipoproteinemia due to familial Lipoprotein Lipase (LPL) deficiency. Diagnosis of persistent hyperlipemic syndromes in kittens should include the possibility of LPL deficiency as determined by PHPLA measurements.     

Ultrastructural alterations of the myocardium of rats fed rapeseed oils.

Ultrastructural studies were conducted on the myocardium of rats fed corn oil, Tower RSO (0 . 88 per cent EA), 1788 RSO (3 . 6 per cent EA) and Target RSO (38 . 9 per cent EA) supplemented diets for 18 weeks. Cardiac myocytes of Tower RSO-fed rats showed some loosening of myofibrils and a slight increase in the number of mitochondria, few of which had lost their cristae. Large intravascular lipid droplets were observed in the myocardium of rats fed the 1788 RSO diet, as well as some small lipid droplets which were seen in close association with mitochondria. There was an apparent increase in the number of mitochondria of both normal and giant size. Many of the mitochondria exhibited distortion of shape and degeneration of cristae. The matrix of megamitochondria contained vesicles and electron-dense floccular inclusions and at times electron-lucent lipid-like material. The degenerative changes of mitochondria were most pronounced in the Target RSO group, where some megamitochondria showed a complete loss of cristae and a replacement of matrix with lipid-like material. These observations suggest that both intravascular lipid globules and the mitochondrial alterations are possible contributory factors involved in the development of cardiac lesions in RSO-fed rats.     

Calcium metabolism in birds.

Calcium is one of the most important plasma constituents in mammals and birds. It provides structural strength and support (bones and eggshell) and plays vital roles in many of the biochemical reactions in the body. The control of calcium metabolism in birds is highly efficient and closely regulated in a number of tissues, primarily parathyroid gland, intestine, kidney, and bone. The hormones with the greatest involvement in calcium regulation in birds are parathyroid hormone, 1,25-dihydroxyvitamin D(3) (calcitriol), and estrogen, with calcitonin playing a minor and uncertain role. The special characteristics of calcium metabolism in birds, mainly associated with egg production, are discussed, along with common clinical disorders secondary to derangements in calcium homeostasis.     

Analysis of complex cytogenetic alterations in three canine mammary sarcomas.

Cells of mammary sarcomas in three dogs were analysed cytogenetically. In an osteoidsarcoma, hyperdiploidy with a range of 92 to 98 chromosomes, and several structural aberrations (for example, a derivative chromosome 1, isochromosome 13 and several bi-armed markers) were observed. Isochromosome 13 was also present in a case of an osteoidchondrosarcoma. In a case of chondro-osteosarcoma both hypodiploidy with a chromosome range of 60 to 66 and hyperdiploidy with a range of 115 to 128 and several centric fusions were observed.     

Ultrastructural alterations in the kidneys of Pekin ducks fed methylmercury.

Ultrastructural alterations in the kidneys of Pekin ducks were studied after subchronic exposure to methylmercury chloride. Twenty-four male and twenty-four female Pekin ducks, 12 weeks of age were fed 0.0 (control), 0.5, 5.0 and 15.0 parts per million of MeHgCl, mixed in a mash diet for 12 weeks. Birds were observed for clinical signs daily. Kidneys were fixed in situ by perfusion with 4% glutaraldehyde in 0.05 M phosphate buffer and routinely processed for electron microscopy. Only minor morphological changes were observed in the renal corpuscle of ducks in the 0.5 ppm group. The component cells of the renal corpuscle in the 5.0 and 15.0 ppm groups showed several ultrastructural alterations, including an increase in number and size of lysosomes and vacuoles in podocytes and mild vacuolation in mesangial cells. The juxtaglomerular cells of afferent arterioles had crystallized granules. No significant changes were observed in glomerular basement membrane thickness and width of podocyte pedicels. The proximal tubules of the 5.0 and 15.0 ppm groups showed some microvillar loss and an increase in the number and size of lipid droplets, vacuoles and lysosomal bodies. The distal tubular changes in the 5.0 and 15 ppm groups included electron lucency and vacuolation of cytoplasm as well as occasional mitochondrial swelling. The collecting ducts of 5.0 ppm and 15.0 ppm groups showed varying degrees of degenerative changes in both mucus secretory as well as absorptive cells. These results suggest that subchronic exposure of Pekin ducks to methylmercury chloride causes subtle ultrastructural changes in renal corpuscle and marked ultrastructural changes in proximal tubules and collecting ducts.     

Plasma protein alterations in cattle suffering from acute peritonitis.

Modification of the normal plasma protein picture has been studied in plasma samples from cows suffering from spontaneously arisen (17 cases) or experimentally induced acute peritonitis (5 cases) using polyacrylamide electrophoresis. Considerable differences were found in the postalbumin region during peritonitis. One protein component increased in size, while another disappeared. Two weak components in normal samples were replaced by four discrete protein bands. These modifications were not detected in any of 10 plasma samples from cows suffering from other diseases than peritonitis or in any of 35 samples from clinically healthy animals. The modifications were visible 12-16 h after injection of the provoking agent and were remarkably alike from one case to the next.