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Biomedical subjects

M Llobera

Publications and source records attributed to M Llobera.

At least 55 records · Page 3Linked to original sources

Synthesis of lipoprotein lipase in the liver of newborn rats and localization of the enzyme by immunofluorescence.

In newborn rats, lipoprotein lipase (LPL) activity was higher in the liver than in several other tissues, such as heart, diaphragm or lungs, and accounted for about 3% of total LPL activity in the body. There was no significant correlation between LPL activity in liver and in plasma. Thus transport of the enzyme from extrahepatic tissues was probably not the major source of LPL in liver. To study LPL biosynthesis directly, newborn rats were injected intraperitoneally with [35S]methionine, and LPL was isolated by immunoprecipitation and separation by SDS/polyacrylamide-gel electrophoresis. Radioactivity in LPL increased with a similar time course in all tissues studied, including the liver. Substantial synthesis of LPL was also demonstrated in isolated perfused livers from newborn rats, whereas synthesis was low in livers from adult rats. There was strong LPL immunofluorescence in livers from newborn rats, mainly within sinusoids and along the walls of larger vessels. This labelling disappeared after perfusion with heparin, which indicates that much of the enzyme is in contact with blood and can take part in lipoprotein metabolism.

Animals↗

High liver lipoprotein lipase activity in hyperlipemic developing rats from undernourished pregnant mothers.

To study the potential relationship between circulating triacylglycerol (TAG) levels and lipoprotein lipase (LPL) activity in the newborn rat liver, pups from undernourished or normal control mothers were nursed by normal dams, and studied at 0, 1, 15 or 30 days of age. Plasma TAG levels and liver TAG concentration increased more in pups from undernourished mothers than they did in controls. At birth, liver LPL activity was similarly high in both groups but, whereas in controls it decreased progressively after birth, in pups from undernourished mothers it remained stable until 15 days of age. Results suggest that the hypertriglyceridemia present in pups from undernourished mothers may be responsible for the sustained high LPL activity in their liver which may enhance the hepatic uptake of circulating TAG.

Animal Nutritional Physiological Phenomena↗

Metabolic response to starvation at late gestation in chronically ethanol-treated and pair-fed undernourished rats.

To study the role of undernourishment in the negative effects of ethanol during pregnancy and to determine whether maternal ethanol intake modifies metabolic response to starvation at late gestation, female rats receiving ethanol in their drinking water before and during pregnancy (ethanol group) were compared with animals that received the same amount of solid diet as the ethanol group rats (pair-fed group) and with normal rats fed ad libitum (control group). All animals were killed on the 21st day of gestation, either in the fed state or after 24-hours fasting. The body weight of ethanol rats was lower than that of controls but higher than that of pair-fed rats. When compared with controls, ethanol and pair-fed rats had reduced fetal body weights, whereas fetal body length was reduced only in the former. In the fed state, blood glucose concentration was lower in the ethanol and pair-fed rats and fetuses than in controls. Twenty-four-hour starvation caused a reduction in this parameter only in control and ethanol mothers. In the fed state, maternal liver glycogen concentration was lower in ethanol and higher in pair-fed mothers than in controls. Blood beta-hydroxybutyrate levels were higher in ethanol-treated mothers than in the others, and 24-hour starvation increased this parameter in ethanol and control rats to a greater extent than in the pair-fed ones. Liver triacylglyceride concentration was higher in ethanol-treated mothers than in the other two groups, and starvation caused this concentration to increase in ethanol and control groups but not in the pair-fed group.(ABSTRACT TRUNCATED AT 250 WORDS)

Alcoholism↗

Morphological recovery of hippocampal pyramidal neurons in the adult rat exposed in utero to ethanol.

Reduced numbers of dendritic spines on the secondary apical dendritic branches and basilar dendrites of CA1 and CA3 pyramidal neurons were observed in ethanol-treated rats during embryonic life aged 15 days when compared with age-matched controls. However, differences were no longer present at the age of 90 days. These results suggest that recovery of some morphological parameters of pyramidal hippocampal neurons may occur in rats exposed in utero to ethanol.

Age Factors↗

Uptake and metabolism of Intralipid by rat liver: an electron-microscopic study.

The metabolism of Intralipid (intravenously injected) was studied in rats fasted for 48 h. At all doses used, the Intralipid triacylglycerols disappeared rapidly from circulation and concomitantly the hepatic content of triacylglycerols and the level of circulating ketone bodies increased, indicating an active metabolism of Intralipid by the liver. To study this possibility further we used an ultrastructural approach. In rats given Intralipid we detected numerous lipid particles in the spaces of Disse, retained in the interdigitations of the hepatocyte. There were also lipid particles attached to the luminal surface of the endothelial cells. Small lipid particles were seen in close contact with endocytic vesicles internalized into hepatocytes but were present mainly in endothelial cells. Inside the endothelial cells, the endocytic vesicles were detected in contact with lysosomes. Inside hepatocytes, a process of sterification seemed to occur in the endoplasmic reticulum as deduced from the presence of small lipid droplets with ill-defined outlines. Large lipid droplets were seen in close contact with mitochondria, indicating a mitochondrial uptake and metabolism of fatty acids to synthesize and release ketone bodies. The possible role of lipoprotein lipase in the liver for the hepatic uptake of Intralipid particles is discussed.

Animals↗

Comparative metabolic effects of chronic ethanol intake and undernutrition in pregnant rats and their fetuses.

Female rats receiving ethanol in the drinking water before and during gestation (ET) were compared to pair-fed animals (PF) and normal controls (C) fed ad libitum. On the 21st day of gestation the maternal body and liver weight, blood glucose, and plasma protein concentrations were lower in ET and PF animals as compared to C. In contrast to C or PF mothers, ET-fed mothers had higher circulating beta-hydroxybutyrate and triacyglyceride levels and beta-hydroxy-butyrate/acetoacetate ratio. Liver triacylglycerides were increased whereas liver glycogen concentration was reduced in ET-fed animals. Only fetal body and liver weights and blood glucose were lower in both ET and PF than in C. Blood beta-hydroxybutyrate was increased and liver glycogen was decreased only in ET fetuses. There were no differences among the groups in fetal circulating beta-hydroxy-butyrate/acetoacetate ratio, plasma proteins, and triacylglycerides or liver triacyglyceride content. Results indicate that certain changes in ET mothers are specifically produced by the ethanol intake rather than undernutrition. Further, metabolic changes occurring in the fetus are influenced by the ethanol effects in the mother and these actions may be added to those directly produced by the ethanol crossing the placenta. However, the collaterals were three times more likely to report more drinking days than the patients; 40.4% (86/213) of the cohabiting contacts reported more drinking days compared to 12.7% (27/213) of the patients reporting more drinking days (p less than 0.001).(ABSTRACT TRUNCATED AT 250 WORDS)

3-Hydroxybutyric Acid↗

Lipoprotein lipase uptake by the liver: localization, turnover, and metabolic role.

We have studied the binding and metabolism of 125I-labeled bovine lipoprotein lipase (LPL) by use of isolated, perfused rat livers. Our data suggest the presence of two types of binding sites, i.e., heparin-sensitive sites that bind primarily the catalytically active form of the lipase and are present at the endothelium in all blood vessels and heparin-insensitive sites that bind both active and inactive forms and are present only within the sinusoids. Forty minutes after uptake by the liver, approximately 50% of the LPL had lost its catalytic activity or been degraded. Three processes were evident: 1) colchicine-sensitive degradation to acid-soluble products, 2) partial proteolysis to fragments similar to those formed by limited digestion with trypsin or plasmin, and 3) a conformational change leading to loss of catalytic activity. Exogenous LPL bound in the liver caused a dramatic increase in the utilization of a perfused triacylglycerol emulsion (Intralipid), with rapid formation of free fatty acids and water-soluble metabolites. When the liver was flushed with heparin, it lost its ability to utilize the fat emulsion. Measurement of the hepatic extraction showed that rat livers take up 100-200 mU endogenous LPL per hour.

Animals↗

Characterization of lipoprotein lipase activity in the newborn rat liver.

Acetone ether powders of livers from starved newborn rats were applied to heparin-Sepharose affinity columns. The so-called hepatic triacylglycerol lipase was eluted with 0.9 M NaCl and a second lipolytic activity peak was eluted with 1.5 M NaCl. The behaviour of this 1.5 M NaCl-eluted fraction against increasing concentrations of serum, NaCl, protamine sulfate and heparin in the assay mixture was almost identical to that shown by partially purified lipoprotein lipase from adult rat adipose tissue, and clearly different from that shown by partially purified hepatic triacylglycerol lipase from the adult rat liver. We conclude that the newborn rat liver contains a lipoprotein lipase activity with similar properties to those found in adult adipose tissue lipoprotein lipase activity. It is suggested that this enzyme enables the neonatal liver to take up circulating triacylglycerols directly.

Adipose Tissue↗

Intralipid administration induces a lipoprotein lipase-like activity in the livers of starved adult rats.

The administration of Intralipid to starved adult rats induces the appearance of lipoprotein lipase (LPL)-like activity in the liver, whereas the so-called hepatic triacylglycerol lipase is unaffected. This LPL-like activity is eluted by 1.5 M-NaCl from heparin-Sepharose columns. This partially purified fraction is inhibited by 1.0 M-NaCl (91%) and by 1.0 mg of protamine sulphate/ml (79%), whereas it is stimulated 69-fold by the presence of 8.0 micrograms of apolipoprotein C-II/ml and inhibited by anti-LPL antibodies. We conclude that Intralipid administration induces the appearance of LPL activity in livers of starved adult rats. Its possible origin is discussed.

Animals↗

Long-term effects of TRH administration on food intake and body weight in the rat.

The effect of long-term TRH administration through drinking water (0.2 mg/ml) on food and water intake and body weight has been studied in three groups of female rats: (1) thyroidectomized, (2) thyroidectomized receiving daily 250 micrograms/kg of L-T4, (3) sham-operated. Treatment with oral TRH for 30 days decreased body weight and increased food intake in sham-operated rats. No TRH effects on body weight or food consumption were observed in either of the other groups of thyroidectomized rats. TRH administration increased circulating T3 levels in sham-operated animals, but had no effects in either hypo- or hyperthyroid, thyroidectomized rats. It can be concluded that the TRH-induced increase on food intake is mediated through the pituitary-thyroid axis.

Animals↗

Ethanol administration in the drinking fluid to pregnant rats as a model for the fetal alcohol syndrome.

Addition of ethanol (ET) to the drinking fluid of pregnant rats has been questioned as an experimental model for the fetal alcohol syndrome (FAS). This model, however, closely simulates human alcohol intake, and in this study we used a modified version of previous protocols to overcome their major defects. A group of female rats was given 10% ET in drinking fluid for one week, 15% for the second week, 20% for the third, and 25% for the fourth, at the end of which they were mated with non-treated males and given 25% ET throughout gestation. Three groups of non-ET treated sex and age-matched rats were studied in parallel: (1) normal controls receiving solid diet ad lib, (2) paired fed rats, and (3) rats fed ad lib the solid diet mixed with 50% fiber. In the ET group, food intake decreased as ET consumption augmented, the ET calories comprising over 30% of the total energy intake during pregnancy. Total energy intake was similar for ET group and normal controls, and was higher than in paired fed animals or those on 50% fiber diet. Body weight gain in ET rats was similar to those on 50% fiber diet, lower than in normal controls and higher than in paired fed animals. At the 21st day of gestation, rats on ET had plasma ethanol levels of 147 +/- 18 mg/dl and higher plasma osmolality than in the other groups studied. In ET rats, fetal body weight was lower than in either normal controls or rats on 50% fiber diet, and fetal body length was shorter than in any other group.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Time-dependent effects of oral thyrotropin-releasing hormone in hyperthyroid female rats.

We have studied the thyrotropin (TSH) and thyroid hormone (T3) response to acute thyrotropin-releasing hormone (TRH) administration in rats previously rendered hyperthyroid by daily ip injection of 25 micrograms L-T4/100 g bw for 30 days. Animals were thereafter put on distilled water or TRH (2 mg/10 ml) as drinking solution for another 18 days, continuing T4 administration. On days 3 and 10 on oral TRH, the serum TSH and T3 response to TRH was studied by administering 2 micrograms of the tripeptide iv. After 3 days, neither the animals on oral DW, nor those on oral TRH showed any response. Nevertheless, after 10 days, animals on oral TRH had a measurable, significant thyrotropin response. At sacrifice (18 days), animals on oral TRH had increased pituitary TSH content. These results confirm and extend a recent report from our laboratory (Iglesias, Llobera and Montoya 1985). We conclude that TRH-mediated thyrotropin accumulation in pituitary is the primary cause for the presence of this response.

Administration, Oral↗

Starvation enhances lipoprotein lipase activity in the liver of the newborn rat.

To determine to what extent lipoprotein lipase activity in the liver of the newborn rat depends on milk ingestion, its changes were studied during different nutritional conditions. Newborns were placed with nurse rats with or without ligated nipples and they were killed at 0,8 or 24 h of life. Lipoprotein lipase in newborns liver was characterized by its inhibition in the presence of 1.0 M NaCl, its specific elution at 1.5 M NaCl on heparin-Sepharose 4B column and its requirement for serum in the assay mixture to manifest its activity. In fed animals lipoprotein lipase activity and triacylglycerol content in liver as well as circulating triacylglycerols and ketone bodies increased progressively after birth. When newborns were kept starved the change in enzyme activity was significantly enhanced, whereas the increase found after birth in the other parameters disappeared. Starvation produced reduction in circulating RIA-insulin levels in the newborn rats. Results show that liver lipoprotein lipase activity in the newborn rat is controlled by a mechanism which resembles that of the enzyme in the adult heart and indicate that its presence facilitates the uptake by the liver of fatty acids from circulating triacylglycerols for their oxidation rather than deposit.

Animals↗

Glucose infused through the portal vein enhances liver gluconeogenesis and glycogenesis from [3-14C]pyruvate in the starved rat.

After a pulse of [3-14C]pyruvate, 24 hr starved rats were infused through the portal vein with two different doses of glucose (7.8 or 20.8 mg/min) or the medium, and blood was collected from the inferior cava vein at the level of the suprahepatic veins. The highest dose of glucose enhanced the appearance of [14C]glucose in blood from the 2nd to the 20th min after tracer delivery. It also enhanced production of [14C]glycogen and concentration of glycogen in the liver after 5 and 20 min. At 20 min of glucose infusion the appearance of [14C]glyceride glycerol in liver as well as liver lactate concentration and lactate/pyruvate ratio were increased. The low dose of glucose used enhanced liver values of [14C]glycogen, [14C]glycogen specific activity and glycogen concentration. Our results support the hypothesis that in the starved rat glucose is converted into C3 units prior to being deposited as liver glycogen and based on the liver zonation model (Jungermann et al., 1983) it is proposed that glucose stimulated gluconeogenesis by shifting the liver to the cytosolic redox state as a secondary consequence of increased glycolytic activity.

Animals↗

Sequential changes in the pituitary thyroid axis after chronic TRH administration: effects on euthyroid and thyroxine treated female rats.

The effect of chronic oral thyrotrophin-releasing hormone (TRH) administration on thyrotrophin (TSH), L-triiodothyronine (T3) and L-thyroxine (T4) serum levels, pituitary TSH concentration and serum response to acute TRH injection, has been studied in female rats under different thyroidal conditions: sham-operated control animals, and thyroidectomized animals receiving 25 micrograms L-T4/100 g body weight/day. After 30 days, these groups were divided into two subgroups (6-10 animals per group), one receiving the aforementioned treatment and the other the same plus 2 mg TRH/10 ml distilled water (DW), as drinking water. TRH-treated sham-operated animals showed significantly reduced serum and pituitary TSH levels and increased serum T3 levels at most of the times studied (1, 6, 10, 18 and 34 days of oral TRH or DW administration), and a transient elevation in serum T4 between day 1 and 6. Thyroidectomized-L-T4-treated animals showed increased serum and pituitary TSH levels throughout the treatment and reduced T3 and T4 serum levels at the beginning, as compared to thyroidectomized-L-T4-treated animals. TSH response to iv TRH administration on the 10th day of oral TRH administration was reduced in controls chronically treated with oral TRH as compared to non-treated controls, and was increased in thyroidectomized-L-T4-treated animals on chronic TRH vs the same group on oral DW. These results suggest that chronic TRH administration can stimulate TRH synthesis in vivo, bypassing the inhibitory effects of thyroid hormones, the increased pituitary TSH reserve being responsible for the partial restoration of a response to acute TRH injection in the thyroidectomized-L-T4-treated animals.

Animals↗

Circulating triacylglycerols, lipoproteins, and tissue lipoprotein lipase activities in rat mothers and offspring during the perinatal period: effect of postmaturity.

Mammary gland and adipose tissue lipoprotein lipase activities have been implicated in the changes of circulating triacylglycerol levels which occur in the mother at late gestation. In the newborn the temporal accumulation of triacylglycerols in the liver coincides with the appearance of a lipoprotein lipase peak. The relationships of these changes with the rise in circulating prolactin in the mother before parturition and the extrauterine nutritional status in the offspring were studied in a postmaturity model produced in the rat by subcutaneous injection of 7 mg progesterone/day to pregnant animals from the 20th day of gestation. Pregnant controls received the medium. Parturition occurred at day 21.5 of gestation in pregnant controls while it did not occur before the 23rd day in those receiving progesterone. At the 20th day of gestation, plasma triacylglycerol concentrations and all lipoprotein fractions (especially VLDL) were much higher in mothers not receiving progesterone than in age-matched virgins, and these differences disappeared at the 21st day of gestation. Lipoprotein lipase activity was maintained low in control mothers' adipose tissue until the 23rd postfecundation day while it greatly increased in mammary gland from parturition time. In progesterone treated mothers, both triacylglycerol and lipoprotein fractions (especially VLDL) in plasma were maintained elevated until the 23rd postfecundation day and adipose tissue and mammary gland lipoprotein lipase activities were maintained low until this time.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Method for triglyceride measurement in small amounts of plasma.

A method is described for determining triacylglyceride concentration in small amounts of plasma. After ethanolic-KOH digestion of diluted plasma aliquots, samples were neutralized with MgSO4 and glycerol was fluorimetrically assayed in supernatants by the coupling of glycerokinase, pyruvate kinase, and lactate dehydrogenase catalyzed reactions. Values were corrected by free glycerol present in the non-digested samples. Digestions were performed at different times and temperatures in order to establish optimum conditions for recoveries and reproducibility. Parallel determinations before and after phospholipid removal showed that their presence in plasma did not interfere with the obtained values. This method is especially useful for running many samples in parallel and for determinations in small experimental animals in which the amount of plasma is very limited.

Fluorometry↗

Morphological and compositional changes of rat plasma triglyceride-rich lipoproteins incubated with adipose tissue.

Triglyceride-rich rat lipoproteins (mainly VLDL), purified by ultracentrifugation and dialysis, were incubated for 120 min in the presence of epididymal fat-pad pieces from fed rats in media containing heparin. During this process, the lipoproteins were depleted of triglycerides and proportionally enriched with proteins, causing their increased density. After incubation, electron microscopic study of the triglyceride-rich lipoproteins revealed partially degraded structures, and some particles had atypical discoid flattened form and marked lamellar structure. These findings are in agreement with those of Blanchette-Mackie & Scow (1976) in triglyceride hydrolysed chylomicrons and confirm their observation of monolayers formed by accumulated lipolytic products that move by lateral diffusion, giving rise to a spiral fold between the core triglycerides and the aqueous space.

Adipose Tissue↗