The effect of stimulation of the mononuclear phagocyte system on the binding of high- and low-density lipoproteins by the hepatocytes, Kupffer cells and liver endotheliocytes of rats

The role of the mononuclear phagocyte system (MPS) in the regulation of low- (LDL) and high-density lipoprotein (HDL) receptor activity in different rat liver cells was investigated. MPS was activated by intravenous administration of bacterial lipopolysaccharide (LPS) from Serratia marcescens. Liver cells were isolated by in vitro perfusion of the liver with a collagenase solution. Separation of Kupffer and endothelial cells was performed by the method of centrifugal elutriation. In control rats, the hepatocytes bound 1.6 times more 125I-HDL and 2.5 times more 125I-LDL per cell than Kupffer cells. Treatment of rats with LPS resulted in a 4.5-fold decrease in the 125I-HDL binding to Kupffer cells. In contrast, the hepatocytes from LPS-treated rat bound 2 times more 125I-HDL than that from untreated rats. The binding of 125I-LDL and 125I-HDL to endothelial cells and 125I-LDL to hepatocytes were not affected by LPS treatment. These results suggest that the MPS (especially Kupffer cells) plays an important role in the regulation of HDL receptor activity in hepatocytes.     

Effect of follicular cells, IGF-I and tyrosine kinase blockers on oocyte maturation

The aim of this study was to test the following hypotheses: (i) that oocyte maturation is controlled by surrounding follicular cells; (ii) that a meiosis-regulating factor of follicular origin is not species-specific; (iii) that one of the follicular regulators of oocyte maturation is IGF-I; and, (iv) that Cumulus oophorus and tyrosine kinase-dependent intracellular mechanisms do not mediate IGF-I action on oocytes. It was found that co-culture of cumulus-enclosed bovine oocytes with isolated bovine ovarian follicles or with isolated porcine ovarian follicles significantly increased the proportion of matured oocytes (at metaphase II of meiosis) after culture. Porcine oocytes without cumulus investments had lower maturation rates than cumulus-enclosed oocytes. Co-culture with isolated porcine ovarian follicles resulted in stimulation of maturation of both cumulus-free and cumulus-enclosed porcine oocytes. These observations suggest that follicular cells (whole follicles or Cumulus oophorus) support bovine and porcine oocyte maturation, and that follicular maturation-promoting factor is not species-specific. The release of significant amounts of IGF-I by cultured bovine and porcine isolated follicles and granulosa cells was demonstrated. Addition of IGF-I to culture medium at 10 or 100 (but not 1000) ng/ml stimulated meiotic maturation of both cumulus-enclosed and cumulus-free porcine oocytes. Neither of the tyrosine kinase blockers, genistein or lavendustin (100 ng/ml medium), changed the stimulating effect of IGF-I on porcine oocytes. The present data suggest that at least one of the follicular stimulators of oocyte nuclear maturation is IGF-I, and that its effect is probably not mediated by cumulus investment or by tyrosine kinase-dependent intracellular mechanisms.     

Innervation of grouped lymphoid nodules (Peyer's patches) by the enteric nervous system and the topography of their interior neural elements in the rat

Using the complex of histological methods (staining with toluidine blue, silver nitrate impregnation and application of retrograde fluorescent dye primulin) the data on the neural elements spatial localization within Peyer patches of the small intestine and their connections with the rest of enteric metasympathetic nervous system in rat was obtained. Submucosal plexus that is significantly developed within this lymphoid organ and is divided into internal and external plexuses was found to be most essential to the innervation of rat Peyers patches. These plexuses innervate all Peyers patch areas:nodules, cupula and internodular zones and the nodule-associated epithelium. Moreover, it was shown that within Peyers patch the plexuses form an integral part with the rest of the enteric nervous system and possess close connections with ganglia that are distant from the patch and are related both to submucosal and myenteric nervous plexuses. Direct inputs into Peyers patch nervous plexuses from extramural ganglia are present as well. These data was considered as a morphological basis for functional interaction of nervous and immune systems within the enteric immune organ and for possible enteric nervous system regulation of immune functions.     

A randomized comparison of two weight-reducing diets. Calorie counting versus low-fat carbohydrate-rich ad libitum diet

We compared the importance of rate of initial weight loss for long term outcome in obese patients and the efficacy of two different dietary weight maintenance programmes. An initial weight loss of 12.6 kg was achieved either by eight weeks low energy diet (2 MJ/day) (n = 21) or 17 weeks conventional hypocaloric, high protein diet (5 MJ/day) (n = 22) both supported by an anorectic compound (ephedrine 20 mg and caffeine 200 mg thrice daily). Weight loss rate had no effect on long-term weight maintenance. Randomisation to one year weight maintenance of either an ad lib, low fat, high carbohydrate diet or a fixed energy diet (< 8 MJ/day), both supported by reinforcement sessions 2-3 times monthly, resulted in a maintenance of 13.2 of initial 13.5 kg weight loss in the ad lib group versus 9.7 of 13.8 kg in the fixed energy intake group. At follow-up two years after the initial weight loss, 65% of the ad lib group and only 40% of the fixed energy intake group had maintained a weight loss of > 5 kg.     

Tomato lycopene and low density lipoprotein oxidation: a human dietary intervention study

Increase in low density lipoprotein (LDL) oxidation is hypothesized to be causally associated with increasing risk of atherosclerosis and coronary heart disease. In recent epidemiological studies, tissue and serum levels of lycopene, a carotenoid available from tomatoes, have been found to be inversely related to risk of coronary heart disease. A study was undertaken to investigate the effect of dietary supplementation of lycopene on LDL oxidation in 19 healthy human subjects. Dietary lycopene was provided using tomato juice, spaghetti sauce, and tomato oleoresin for a period of 1 wk each. Blood samples were collected at the end of each treatment. Serum lycopene was extracted and measured by high-performance liquid chromatography using an absorbance detector. Serum LDL was isolated by precipitation with buffered heparin, and thiobarbituric acid-reactive substances (TBARS) and conjugated dienes (CD) were measured to estimate LDL oxidation. Both methods, to measure LDL oxidation LDL-TBARS and LDL-CD, were in good agreement with each other. Dietary supplementation of lycopene significantly increased serum lycopene levels by at least twofold. Although there was no change in serum cholesterol levels (total, LDL, or high-density lipoprotein), serum lipid peroxidation and LDL oxidation were significantly decreased. These results may have relevance for decreasing the risk for coronary heart disease.     

Domain structure analysis of elongation factor-3 from Saccharomyces cerevisiae by limited proteolysis and differential scanning calorimetry

Elongation-factor-3 (EF-3) is an essential factor of the fungal protein synthesis machinery. In this communication the structure of EF-3 from Saccharomyces cerevisiae is characterized by differential scanning calorimetry (DSC), ultracentrifugation, and limited tryptic digestion. DSC shows a major transition at a relatively low temperature of 39 degrees C, and a minor transition at 58 degrees C. Ultracentrifugation shows that EF-3 is a monomer; thus, these transitions could not reflect the unfolding or dissociation of a multimeric structure. EF-3 forms small aggregates, however, when incubated at room temperature for an extended period of time. Limited proteolysis of EF-3 with trypsin produced the first cleavage at the N-side of Gln775, generating a 90-kDa N-terminal fragment and a 33-kDa C-terminal fragment. The N-terminal fragment slowly undergoes further digestion generating two major bands, one at approximately 75 kDa and the other at approximately 55 kDa. The latter was unusually resistant to further tryptic digestion. The 33-kDa C-terminal fragment was highly sensitive to tryptic digestion. A 30-min tryptic digest showed that the N-terminal 60% of EF-3 was relatively inaccessible to trypsin, whereas the C-terminal 40% was readily digested. These results suggest a tight structure of the N-terminus, which may give rise to the 58 degrees C transition, and a loose structure of the C-terminus, giving rise to the 39 degrees C transition. Three potentially functional domains of the protein were relatively resistant to proteolysis: the supposed S5-homologous domain (Lys102-Ile368), the N-terminal ATP-binding cassette (Gly463-Lys622), and the aminoacyl-tRNA-synthase homologous domain (Glu820-Gly865). Both the basal and ribosome-stimulated ATPase activities were inactivated by trypsin, but the ribosome-stimulated activity was inactivated faster.