Effects of thyroxin therapy on different analytes related to obesity and inflammation in dogs with hypothyroidism.

Hypothyroidism in dogs is accompanied by changes in intermediary metabolism including alterations in bodyweight (BW), insulin resistance, and lipid profile. In this study, changes in selected adipokines (adiponectin, leptin), butyrylcholinesterase (BChE), and acute phase proteins, including C-reactive protein, haptoglobin (Hp) and serum amyloid A (SAA), were studied in dogs with hypothyroidism under thyroxin therapy. Blood samples were collected when hypothyroidism was diagnosed (before treatment) and after treatment with thyroxin. Twenty-eight of 39 dogs exhibited a good therapeutic response (group A), whereas the remainder were considered to have been insufficiently treated (group B). Following treatment, group A dogs demonstrated a statistically significant decrease in canine thyroid stimulating hormone (c-TSH) (P<0.001) and an increase in free thyroxine (fT4) (P<0.001) concentrations, associated with a significant decrease in BW (P<0.05), leptin (P<0.01), and adiponectin, (P<0.001) and an increase in BChE (P<0.01) and Hp (P<0.05). Group B dogs showed no statistically significant changes in c-TSH, but had a significant increase in fT4 (P<0.001) accompanied by a significant decrease in adiponectin (P<0.05) of lower magnitude than group A. No significant changes in the mean circulating levels of APPs were observed in both groups, with the exception of an increase in Hp (P<0.05) in group A. In summary, the successful treatment of hypothyroidism reduces circulating levels of adiponectin and leptin, while increasing BChE activity in dogs. The mean increase in Hp values and decrease in SAA for some of the dogs after treatment warrants further investigation.

Variations of plasma leptin in show horses during a work season.

Leptin is an adipocytokine mainly expressed by adipose tissue. Secretion of leptin in healthy animals is closely related to fat mass and metabolic activity. The aim of this study was to investigate plasma leptin variations, in relation to nutritional and exercise parameters in adult show horses during a work season. EDTA-blood samples were taken at rest from 37 Iberian horses. Body weight, body condition score and fat percentage determined by ultrasonic measurement of rump fat thickness were measured. Plasma leptin was determined with a multi-species RIA kit. Linear mixed effects model was used to assess relationship between plasma leptin and other biological parameters. Plasma leptin concentration was <12.6 ng/ml (mean = 2.8 ± 1.6 ng/ml) and was significantly higher during training periods (p < 0.0001) (4.5 ± 1.7 ng/ml) than in show periods (2.0 ± 1.1 ng/ml), despite a significant increase (p < 0.0001) in energy intake. The body weight remained almost constant. The plasma leptin concentrations were significantly affected by exercise (p < 0.0001), body weight (p = 0.04) and BCS (p < 0.0001), but were not affected by percentage of fat. In conclusion, the marked decrease in leptin values observed during a period of intense (i.e. excessive) exercise could result from an adaptation to cumulative alterations in energy balance, to exercise per se or to a combination of both.

Liver lipid metabolism.

The liver plays a key role in lipid metabolism. Depending on species it is, more or less, the hub of fatty acid synthesis and lipid circulation through lipoprotein synthesis. Eventually the accumulation of lipid droplets into the hepatocytes results in hepatic steatosis, which may develop as a consequence of multiple dysfunctions such as alterations in beta-oxidation, very low density lipoprotein secretion, and pathways involved in the synthesis of fatty acids. In addition an increased circulating pool of non-esterified fatty acid may also to be a major determinant in the pathogenesis fatty liver disease. This review also focuses on transcription factors such as sterol-regulatory-element-binding protein-1c and peroxisome proliferator-activated receptor alpha, which promote either hepatic fatty acid synthesis or oxidation.

Hormonal disturbances associated with obesity in dogs.

Obesity is associated with multiple endocrine alterations and changes in the concentration of circulating hormones. However, few studies have explored such alterations in dogs with naturally acquired excess weight. In the present study, we investigated the effect of naturally acquired obesity on cortisol, insulin-like growth factor (IGF)-1 and prolactin secretion in dogs. Thirty-one overweight dogs were enrolled in the trial. Blood samples were collected before and after adrenocorticotrophic hormone (ACTH) injection. Free thyroxine (fT4), cortisol, thyroid-stimulating hormone (TSH), IGF-1, prolactin and fructosamine were assayed. Body weight excess increased significantly with age and neutered dogs were more obese than entire ones. The ACTH stimulation test was within the normal range for 26 of 31 dogs. Prolactinaemia was increased in seven dogs and IGF-1 in six dogs. Twenty dogs had a fructosamine concentration >340 microm. Interestingly, 18 of 31 dogs showed disturbances of thyroid function based on high TSH and/or low fT4 baseline concentration, with 11 dogs showing both. According to these parameters only six of 31 dogs were free of hormonal disturbances. These results revealed the high incidence of such disturbances, especially thyroid dysfunction, in obese, but otherwise apparently healthy dogs. They demonstrate the importance of examining endocrine function during the initial evaluation of obese dogs to avoid failure of any nutritional treatment.

Childhood obesity and insulin resistance in a Yucatan mini-piglet model: putative roles of IGF-1 and muscle PPARs in adipose tissue activity and development.

OBJECTIVE: To explore metabolic and cellular modifications induced during childhood obesity, in a novel animal model of obese mini-piglets. DESIGN: A total of 10 four-month old Yucatan mini-pigs were followed from prepuberty to adulthood. Animals were divided into two groups. The first one had been overfed (OF) a western-type diet and the second one had been normally fed a control recommended human-type diet (NF). MEASUREMENTS: Plasma insulin-like growth factor 1 (IGF-1), insulin, leptin, nonesterified fatty acids, triglycerides (TGs) and glucose were determined at sexual maturity and at young adulthood. Quantitative gene expressions of peroxysome-proliferator-activated receptors (PPARs), glucose transporter 4, insulin receptor, IGF-1, leptin and interleukin-6 (IL-6) in skeletal muscle, adipose tissue and liver were also measured at both stages. Adult insulin sensitivity was measured via euglycaemic-hyperinsulinaemic clamps. RESULTS: Increased body weight in adult OF pigs was associated with increased body size and low insulin sensitivity. Sexually mature OF pigs had higher IGF-1 plasma concentrations than their lean littermates (P < 0.05). In the OF group, TGs and glucose were both decreased (P < 0.05). Muscle PPARgamma and alpha in OF pubescent pigs as compared to NF pigs were 11 times higher and 20 times lower, respectively (P < 0.01). CONCLUSION: Obesity and insulin resistance induced by overfeeding mini-pigs during development and puberty were not associated with the cluster of metabolic modifications frequently observed in their adult littermates. Increased IGF-1 concentrations and modifications of skeletal muscle PPAR (alpha and gamma) expressions may help the young obese pig to partially regulate its glycaemia and triglyceridaemia through an increase of fat mass, which maintains its high insulin sensitivity.

Insulin resistance and changes in plasma concentration of TNFalpha, IGF1, and NEFA in dogs during weight gain and obesity.

Obesity-induced insulin resistance (IR) is a common problem in humans as well as domestic dogs. It is well-known that this syndrome is associated with many modifications but it is still unclear if the changes are alterations or adaptations. The purpose of this study was to develop obesity-induced IR in dogs, through a long-term overfeeding period, and to explore hormonal and metabolic disturbances associated with the development of this syndrome. Dogs were overfed for 7 months. Body weight increased by 43 +/- 5%, and insulin sensitivity decreased by 44 +/- 5%. Plasma insulin-like growth factor 1 (IGF1), tumour necrosis factor alpha (TNFalpha), and non-esterified fatty acids (NEFA) concentrations progressively increased during the overfeeding period (IGF1: 111 +/- 13 to 266 +/- 32 ng/ml, p < 0.001; TNFalpha: 5 +/- 5 to 134 +/- 41 pg/ml; NEFA: 0.974 +/- 0.094 to 1.590 +/- 0.127 mmol/l, p < 0.05). These metabolic and hormonal impairments are associated with IR, in obese dogs, and could explain, at least in part, the outbreak of this syndrome.

An insulin-resistant hypertriglyceridaemic normotensive obese dog model: assessment of insulin resistance by the euglycaemic hyperinsulinaemic clamp in combination with the stable isotope technique.

Many studies have shown that in humans insulin resistance (IR) is associated with obesity and hypertriglyceridaemia. The aim of our study was to develop slowly dietary-induced obesity in dogs through long-term overfeeding of a high-fat diet, and to characterize this IR, hypertriglyceridaemic and normotensive model. Insulin resistance was assessed by the euglycaemic hyperinsulinaemic clamp technique. The contribution of hepatic glucose production during the clamp was evaluated using a constant stable-isotope-labelled glucose infusion. Overfeeding a high-fat diet for 7 months was associated with a 43+/-5% body weight increase. Insulin resistance was characterized by hyperinsulinaemia in the unfed state (10+/-1 vs. 24+/-1 microU/ml, in healthy and obese dogs, respectively, p<0.02) and by a reduction of the insulin-mediated glucose uptake (28+/-3 vs. 16+/-1 mg/kg/min, p<0.02). Hepatic glucose production suppression under insulin infusion allowed to conclude that this reduced glucose uptake resulted from a decrease of insulin sensitivity in obese dogs. Furthermore, animals remained normotensive and exhibited a marked hypertriglyceridaemia (0.26+/-0.04 vs. 0.76+/-0.15 mmol/l, in healthy and obese dogs, respectively, p<0.02). Because hypertriglyceridaemia is the most common lipid abnormality in insulin-resistant humans, this dog with slowly induced obesity may constitute a good model to study the consequences of IR in lipid metabolism independently of vascular changes.

Leptin, body fat content and energy expenditure in intact and gonadectomized adult cats: a preliminary study.

The present study was conducted to assess the body composition, leptin, and energy expenditure changes following gonadectomy in cats. Twenty-one females (12 intact and nine spayed) and 21 males (11 intact and 10 castrated) were used. Body weight was recorded. Serum plasma leptin was measured by radioimmunoassay and body composition and energy expenditure were assessed after injection of doubly labelled water. These results confirmed the gain in body weight and body fat following neutering and demonstrated a strong linear relationship between body fat and serum level of leptin. Energy expenditure decreased in castrated cats in comparison with intact ones. This study underlined the effect of gonadectomy as a major factor of obesity in cats and showed that the increase in circulating leptin reflected the amount of body fat. The present results provide further evidence that the regimen of gonadectomized cats should be carefully controlled to avoid excessive weight gain.

Comparison of two protocols with a progesterone antagonist aglepristone (RU534) to induce parturition in bitches.

Parturition was induced in ten Beagle bitches by injecting them subcutaneously with 15 mg aglepristone kg-1 (Alizine) at day 58 of gestation and 24 h later and subsequently at 2 h intervals with either 0.08 mg alfaprostol kg-1 (Alfabedyl) (group 1; five bitches) or 0.15 iu oxytocin kg-1 (Ocytocine S) (group 2; five bitches). Blood samples were collected every 4 h until the end of parturition to assay plasma concentrations of progesterone, dihydro-keto prostaglandin F2 alpha (PGFM), oxytocin, prolactin and cortisol. Parturition occurred in all bitches. The mean time of onset of parturition for both groups was not significantly different (32.6 +/- 3.7 h for group 1 versus 31.6 +/- 3.6 h for group 2), although the mean expulsion time for bitches from group 2 (4.5 +/- 1.8 h) was significantly shorter than that of bitches in group 1 (9.1 +/- 2.0 h). At birth, 93% of the pups were alive in group 2 compared with 86% in group 1. Peripheral plasma concentrations of progesterone increased significantly after the administration of aglepristone, but direct or indirect luteolysis was not induced, and plasma concentrations of oxytocin or cortisol did not change during the first 24 h after administration of aglepristone. PGFM concentrations increased significantly after 4 h of aglepristone administration. During the first 20 h after aglepristone administration, prolactin concentrations increased significantly. At parturition, bitches in group 2, which had the shorter expulsion time of pups, were characterized by significantly higher concentrations of oxytocin and PGFM than bitches in group 1.

Hormonal variation in bitches after early or mid-pregnancy termination with aglepristone (RU534).

Seven bitches in early pregnancy (12.8 +/- 3.8 days after ovulation; group 1) and seven bitches in mid-pregnancy (32.0 +/- 1.53 days after ovulation; group 2) were used in this study. For each group, five bitches were treated with 0.10 mg aglepristone (Alizine) kg-1 and this dose was repeated 24 h later. Two control bitches received a placebo. Blood samples were collected at 6 h intervals to determine plasma concentrations of progesterone, dihydro-keto prostaglandin F2 alpha (PGFM), oxytocin, prolactin and cortisol. Parturition occurred in the four control bitches. All bitches treated with aglepristone aborted. In group 1, embryonic death occurred; in group 2, fetal expulsion occurred 60-132 h after administration of aglepristone. After pregnancy termination, the interoestrous interval of aglepristone-treated bitches was significantly shorter than that before treatment. Treatment with aglepristone did not modify plasma concentrations of progesterone, prostaglandin, oxytocin or cortisol within 24 h after its administration, but it induced, in mid-pregnancy (group 2) a discharge of prolactin within 12 h after its administration. As an abortifacient, aglepristone acted on the uterus and, therefore, did not have direct or immediate luteolytic properties. Termination of pregnancy occurred with high plasma progesterone concentrations. Fetal expulsion was characterized by an increase in the concentration of PGFM, but oxytocin and cortisol remained at basal concentrations.

Quantitative histological analysis of equine embryos at exactly 156 and 168 h after ovulation.

Equine embryos were collected at exactly 156 +/- 0.5 (n=8) and 168 +/- 0.5 h (n=11) after ovulation. The embryos were fixed in glutaraldehyde, sectioned serially and observed using light microscopy. In the 156 h group, all embryos were early blastocysts except for one, which was a morula. The morula and one early blastocyst had no capsule. The capsules of the other embryos were thin. The mean +/- SD total number of cells was 275 +/- 105 (range 117-417). The mean +/- SD proportions of mitotic and pycnotic cells were 2.5 +/- 1.2 and 1.1 +/- 1.8%, respectively, and there were no differences between each inner cell mass (ICM) and trophoblast. The mean +/- SD proportion of ICM cells was 36.5 +/- 5.2%. In the 168 h group, there were early, mid- and expanded blastocysts, all of which had a capsule. The mean +/- SD total number of cells was 1093 +/- 666 (range 272-2217). The mean +/- SD proportions of mitotic and pycnotic cells were 3.5 +/- 1.4% and 0.1 +/- 0.03%, respectively, and there were no differences between each ICM and trophoblast. The mean +/- SD proportion of ICM cells was 21.1 +/- 9.7%. In the 168 h group, there was a significant correlation (P < 0.01) between the total number of cells and the diameters and proportions of ICM cells. The 168 h embryos were composed of significantly (P < 0.01) more cells (approximately four times) than were the 156 h embryos but had lower proportions of ICM cells. These results indicate that in equine embryos there is: (i) an individual (perhaps seasonal) variability in the rate of development; (ii) a doubling in the number of cells every 6 h between 156 h and 168 h after ovulation; and (iii) a decrease in the proportion of ICM cells between the early and expanded blastocyst stages of development.

Cholesterol biosynthesis inhibited by BM15.766 induces holoprosencephaly in the rat.

To confirm that blocking 7-dehydrocholesterol delta 7 reductase (7DHC reductase), as observed in Smith-Lemli-Opitz syndrome (SLOS), induces craniofacial defects, we tested BM15.766, which blocks 7DHC reductase but is chemically unrelated to the holoprosencephaly-inducing teratogen AY9944. Rats were given BM15.766 either in methylcellulose from days (D) 1 through D11 (3 treated groups: protocol A) or in olive oil from D4 through D7 (300 mg/kg/d: protocol B). The sera were sampled on D0, D3, and D5 or D6, D10, D14, and D21 to measure cholesterol and dehydrocholesterols in all groups and steroid hormones in protocol B. D21 fetuses showed the holoprosencephaly spectrum of malformations and the treated dams low cholesterol and accumulation of 7DHC, 8DHC, and trienols, as in SLOS-affected children. In the 3 dosage groups the malformations were dose-related and enzymatic cholesterol decreased to a plateau. The DHC reached 25-44% of the total sterols in the dams. In protocol B, one-third of the BM15.766-treated fetuses presented facial malformations and almost two-thirds pituitary agenesis. On D10, cholesterol reached a minimum and the DHC a maximum while estradiol 17 beta and progesterone were lowered, the latter decreasing in correlation with cholesterolemia. A sterol profile similar to that previously observed after AY9944 associated with a similarly high incidence of pituitary agenesis confirmed that time-limited inhibition of 7DHC reductase induces holoprosencephaly and that pituitary agenesis is the minor form of holoprosencephaly.

Inhibition of 7-dehydrocholesterol reductase by the teratogen AY9944: a rat model for Smith-Lemli-Opitz syndrome.

Our aim is to verify the validity of a rat model proposed for Smith-Lemli-Opitz (SLO) syndrome, a developmental disorder characterized by a defect in 7-dehydrocholesterol-delta 7 (7DHC)-reductase and by facial dysmorphism close to the holoprosencephaly caused by the teratogen AY9944. We investigated the sterol profile in rats treated with AY9944 blocking 7DHC-reductase. AY9944 was given orally to rats on gestation day 3 (D3). The sera were sampled for kinetic data on D3, D6, D9, D12, and D21. Cholesterol was measured in parallel by the routine enzymatic method and by the gas chromatography/mass spectrometry (GC-MS) procedure used in SLO diagnosis. In addition to sterols, we dosed steroid hormones punctually on D4 and on D10, and examined D21 fetuses in other animals. The enzymatic method was not specific for cholesterol, and measured 70% pure 7DHC added to a normal serum. On D21, 77% live fetuses showed pituitary agenesis. Cholesterol was rapidly reduced by more than 50% on D6 involving an accumulation of 7DHC, 8DHC, and trienols, as identified in SLO-affected children. The most abundant 7DHC reached a maximum from D9 to D12, equaling cholesterol on D9 (11 mg/dl). On D10, the magnitudes of hypocholesterolemic and of 7DHC accumulation were found to be dose-dependent. Progesterone was reduced as early as 24 hr after treatment and dropped to 40% of the levels in the controls on D10, correlating to the decrease in cholesterolemia. This rat model reproduces the same biochemical perturbations as seen in SLO, strongly suggesting that aberrant sterols (7DHC, 8DHC, or nortrienol) may contribute to the developmental defects.

Laboratory diagnosis of insulinoma in the dog: a retrospective study and a new diagnostic procedure.

Simultaneous insulin and glucose measurements from a single sample are often insufficient for the laboratory diagnosis of insulinoma in the hypoglycaemic dog. Although the simple insulin (microU/litre): glucose (mmol/litre) ratio is best, when the selected threshold is set at 13.5, it is not always efficient to confirm or refute insulinoma. The fasting test consists of collecting four samples on a given day (no accurate time period is set between any two samples) from a fasting dog for simultaneous insulin and glucose measurements to detect at least one abnormal insulinaemia peak.

Fertilization time in the bitch in relation to plasma concentration of oestradiol, progesterone and luteinizing hormone and vaginal smears.

Seven bitches of several breeds were monitored during oestrus by vaginal smears and luteinizing hormone (LH), oestradiol and progesterone plasma assays. Each bitch was inseminated into the uterus with frozen spermatozoa from seven dogs of different breeds. Each female was inseminated daily with about 200 x 10(6) motile spermatozoa. Six bitches delivered. The paternity of puppies (which indicated the fertilization time) was determined by morphological analysis. Two bitches were fertilized with one artificial insemination (AI) only and four bitches with two successive AIs. Fertilization occurred 84-180 h after the LH peak and 1-3 days before the first day of metoestrus. The gestation length, from fertilization to birth, was 58-63 days or 56-60 days after the first day of metoestrus.

[Midterm toxicity of nitrates: experimental evaluation of the effects on reproductive functions in the female rabbit]. / Toxicité à moyen terme des nitrates: évaluation expérimentale des effets sur les fonctions de reproduction chez la lapine.

Chronic exposure to low levels of nitrate are associated with detrimental effects on reproduction in animals. In order to determine this relationship, an experiment was conducted: female rabbits were given nitrate doses of 0, 250 or 500 mg/l in drinking water for several months. After 2 successive gestations, no detrimental effect was observed on fertility, litter size or weight at birth and weaning, or on plasma oestradiol and progesterone concentration. Hemoglobin level was slightly decreased in dams given 500 mg/l.