Species-dependent stereospecific serum protein binding of the oral anticoagulant drug phenprocoumon.

13 mammalian species are classified into 3 clearcut groups with respect to the stereospecific serum protein-binding of phenprocoumon: 2 groups showing opposed stereospecific binding characteristics and a 3rd group exhibiting no stereospecific binding. Structural differences in the albumin molecule account for these stereospecific differences in serum protein-binding.     

Opiate receptor sites in neuronal and glial cell lines in culture]

A stereospecific saturable high affinity binding of 3H-naloxone has been found in 3 glial and 2 neuronal cell lines. Kinetic study of binding seems to indicate only one class of receptor sites in the 5 cell lines. Acute exposure to morphine (1 X 10(-5)M) concurrent with PGE1-induced stimulation of adenylate cyclase did not result in a decrease of the cAMP level in any cell line tested.     

3[H]-Sulpiride labels mesolimbic non-dopaminergic sites that bind antidepressant drugs

3[H]-(-)-Sulpiride and 3[H]-spiperone binding was compared in rat amygdala, nucleus accumbens and striatum, using (+/-)-sulpiride to define specific binding. 3[H]-(-)-Sulpiride bound to twice as many sites in amygdala and nucleus accumbens as 3[H]-spiperone. 3[H]-(-)-Sulpiride binding was directed to these additional sites by using 1 microM spiperone to mask dopaminergic binding. The binding of 3[H]-(-)-sulpiride to these sites was high affinity, reversible, Na+-dependent, but not stereospecific. Metoclopramide, tiapride and antidepressant medications, but not other neuroleptics, ADTN, or serotonin displaced 3[H]-(-)-sulpiride binding to these sites. These data suggest that 3[H]-(-)-sulpiride labels mesolimbic sites other than dopamine receptors which may mediate antidepressant effects.     

Microbiological reduction of steroidal ketones using the thermophilic bacteriumCaldariella acidophila

Summary Twelve steroidal ketones have been subjected to reduction withC. acidophila resting cells, regio- and stereospecific reduction of the 3-keto groups being observed, as well as reduction of the ⁴-double bond. The presence of oxo groups at C-11 or C-12 and the presence of hydrophobic side chains on the steroidal molecules inhibit the reduction.     

Sur l'interaction entre aldéhydes ou réductones et acides aminés ou protéines

Summary Proteins were exposed to acetaldehyde vapour or mixed with glucose in solution and the basic groups determined by their dy binding capacity. Only serum albumin shows a marked decrease of basic groups and of the carbon/nitrogen ratio as a sign of condensation of aldehyde and amine groups. These results were corroborated by the direct estimation of the aldehyde resine formed on the protein: serum albumin and fibrin treated with acetaldehyde vapour contained 5 to 10 times more resin than the other proteins examined. No rapid combination between serum proteins and glucose could be demonstrated by the dy method or by the kinetics of change of pH. The slow reaction seems not to be dependent on denaturation.     

Glucose does not influence the insulin-like growth factor (IGF) binding to carrier proteins (IGFBPs): Analysis of rat and human serum by western ligand blotting

The insulin-like growth factors (IGFs) circulate bound to specific proteins (termed IGFBP-1 through IGFBP-6) that modulate IGF bioactivity in tissues. The aim of this study was to analyse the effects of glucose on IGF binding to IGFBPs in rat and human serum by means of western ligand blotting. Serum samples were incubated with increasing concentrations of glucose (0 to 50 mmol/l), and EDTA (25 mmol/l) was added to inhibit protease activity. To analyse the effect of glucose on protection of IGFBPs from protease activity, serum from pregnant women (reported to be very rich in proteolytic activity against IGFBPs) was added to rat serum previously incubated with glucose. Glucose did not affect the¹²⁵I-IGF binding to rat and human serum IGFBPs. The intensity of IGFBP-3 bands decreased considerably during the incubation. This appeared to be due to endogenous protease activity, since the decrease was blocked by addition of EDTA. The incubationi of rat serum with pregnant human serum produced a marked attenuation of IGFBP-3 and disappearance of IGFBP-4 bands. In conclusion, our study shows that glucose does not influence the IGF binding to IGFBP-3 either in rat or in human serum, confirms the presence of endogenous proteolytic activity in normal non-pregnant serum, and demonstrates that glucose has no protective action against protease activity.     

Binding property of rat and Limulus C-reactive proteins (CRP) to mercury

The C-reactive proteins (CRP) from both rat and Limulus were found to bind mercury (Hg) in both in vivo and in vitro conditions. CRP has high-affinity binding sites for Hg as evidenced by the loss of free sulfhydryl groups, arrested mobility in polyacrylamide gel electrophoresis, and the consumption of CRP in the serum after Hg administration. The binding was tight as it could not be inhibited either by the addition of cysteine or EDTA. By using a direct titration method it was shown that binding of Hg to CRP was saturable at a molar ratio of Hg/CRP = 13.11. The possibility that CRP may act as a scavenger for Hg is discussed.     

Species-specific differences in the mitogenic activity of heparin-binding growth factors in the sera of various mammals

Sera from different mammalian species displayed great differences in mitogenic activity, as measured by stimulation of DNA synthesis in BALB/c 3T3 cells (3T3 cells). Among the sera examined, fetal bovine serum was least active, and increasing activity was detected in calf serum, human serum, rat serum and mouse serum, in that order. Rat and mouse sera exhibited extremely high mitogenic activity with 3T3 cells, but when TIG-1 human fetal lung fibroblasts were used for the DNA assay instead, the activity levels of all of the sera were lower, and the differences between them were smaller. To determine the reasons for these differences, the heparin-binding growth factors in each serum were separated on a heparin affinity column. Five peaks of DNA-stimulating activity were obtained. Three of these were found in all sera examined, with both 3T3 cells and TIG-1 cells. Two other peaks were found only with 3T3 cells; one was peculiar to rat and mouse sera, with extremely high activity in the rat, and the other was specific to fetal serum. The dependence of the activity of these peaks on the cells used for the test was confirmed using normal rat lung fibroblasts and immortalized rat kidney cells. These findings adequately explain the species-specific differences in mitogenic activity of whole sera, and the variation in activity depending on the cells used for assay of DNA synthesis.     

Species-specific differences in the mitogenic activity of heparin-binding growth factors in the sera of various mammals.

Sera from different mammalian species displayed great differences in mitogenic activity, as measured by stimulation of DNA synthesis in BALB/c 3T3 cells (3T3 cells). Among the sera examined, fetal bovine serum was least active, and increasing activity was detected in calf serum, human serum, rat serum and mouse serum, in that order. Rat and mouse sera exhibited extremely high mitogenic activity with 3T3 cells, but when TIG-1 human fetal lung fibroblasts were used for the DNA assay instead, the activity levels of all of the sera were lower, and the differences between them were smaller. To determine the reasons for these differences, the heparin-binding growth factors in each serum were separated on a heparin affinity column. Five peaks of DNA-stimulating activity were obtained. Three of these were found in all sera examined, with both 3T3 cells and TIG-1 cells. Two other peaks were found only with 3T3 cells; one was peculiar to rat and mouse sera, with extremely high activity in the rat, and the other was specific to fetal serum. The dependence of the activity of these peaks on the cells used for the test was confirmed using normal rat lung fibroblasts and immortalized rat kidney cells. These findings adequately explain the species-specific differences in mitogenic activity of whole sera, and the variation in activity depending on the cells used for assay of DNA synthesis.     

Binding property of rat andLimulus C-reactive proteins (CRP) to mercury

Summary The C-reactive proteins (CRP) from both rat andLimulus were found to bind mercury (Hg) in both in vivo and in vitro conditions. CRP has high-affinity binding sites for Hg as evidenced by the loss of free sulfhydryl groups, arrested mobility in polyacrylamide gel electrophoresis, and the consumption of CRP in the serum after Hg administration. The binding was tight as it could not be inhibited either by the addition of cysteine or EDTA. By using a direct titration method it was shown that binding of Hg to CRP was saturable at a molar ratio of Hg/CRP=13.11. The possibility that CRP may act as a scavenger for Hg is discussed.     

The binding of polyphenols (rutin and some of itsO-β-hydroxyethyl derivatives) to human serum proteins

Summary A binding of polyphenols (rutin and its o--hydroxyethyl derivatives) to human serum was demonstrated. The results showed an increase of binding proportional to the number of free phenolic groups on the molecule of rutin.Acknowledgments. We should like to thank MissC. Gaillard and MissE. Engels for their valuable assistance in this study.     

Potentiation of immunologic memory by Corynebacterium parvum and its mechanism]

Immune response to bovine serum albumin (BSA) at dose of 2,50 mg/kg which is rather a weak immunogen in Rabbits, when given intravenous was highly potentiated when the animals received a previous single intravenous infection of 2 mg/kg of C. parvum, followed by subsequent BSA anamnestic challenges for several months. Thus, the antibody amounts synthesized following the 1st anamnestic injection (3 weeks) were 0,260 mg/ml in the control versus, 0,800 mg/ml in the C. parvum pretreated groups; following the 2nd anamnestic challenge (12 weeks afterwards) 1 mg/ml in the control versus, 2,50 mg/ml in the treated groups following the 3rd anamnestic challenge (28 weeks afterwards) 1,3 mg/ml in the control versus 5 mg/ml in the C. parvum pretreated groups; following the 4th anamnestic challenge (52 weeks afterwards) 0,300 mg/ml in the control versus 0,800 mg/ml in the C. parvum treated groups. On the whole for the four first anamnestic challenges the differences at peak levels between the control and C. parvum treated groups were about to 4. Furthermore, the antibody molecules synthesized by the C. parvum treated animals were found to belong to IgG class. The results suggest that the immunological mechanisms mobilized are peculiar to C. parvum since they could not be reproduced either by BCG or by Freun'd adjuvant under similar conditions.     

Binding of 3,3',5'-triiodo-L-thyronine (rT3) to human serum proteins]

The rT3-binding and human serum proteins was directly studied with tracer doses of radioactive rT3. Polyacrylamide gel electrophoresis showed 125I-rT3 added to human serum was distributed among two proteins: albumin (carrying 57% of tracer rT3) and TBPA )22%). No binding was observed to TBG, protein binding T4.     

A 30-kDa fragment of insulin-like growth factor (IGF) binding protein-3 in human pregnancy serum with strongly reduced IGF-I binding

Proteolytic cleavage of insulin-like growth factor (IGF) binding protein (IGFBP)-3 during pregnancy is likely to have both IGF-dependent and -independent effects on maternal, placental and fetal growth and metabolism. A 30-kDa proteolytic IGFBP-3 fragment was isolated from third trimester pregnancy human serum and identified by N- and C-terminal amino acid sequence analysis and mass spectrometry to correspond to residues 1–212 of the parent protein. This fragment is the dominating IGFBP-3 immunoreactive species in pregnancy serum. The 30-kDa fragment was also detected in serum of non-pregnant women where it coexists with intact IGFBP-3. Using biosensor technology, (1–212)IGFBP-3 was found to have 11-fold lower affinity for IGF-I compared to intact IGFBP-3, while a 4-fold decrease in affinity was found for IGF-II. Tests with des(1–3)IGF-I suggest fast binding of IGF-I to the N-terminal region of IGFBP-3 and similar affinity to a slow binding site in the C-terminal region. Received 24 April 2007; received after revision 11 June 2007; accepted 13 June 2007     

Role of serum components in the binding and phagocytosis of oxidatively damaged erythrocytes by autologous mouse macrophages

To investigate the role of autologous serum components in the recognition of damaged cells by macrophages, we examined the binding and phagocytosis of damage oxidatively damaged red blood cells with Cu2+ and ascorbate (oxRBCs) by autologous resident mouse peritoneal macrophages. The binding of oxRBCs by macrophages was independent of the presence of serum. However, phagocytosis by macrophages increased with serum concentration, and macrophages showed little ingestion of oxRBCs in a serum-free medium. Macrophages neither bound nor appreciably ingested native RBCs (before oxidation) in either the absence or presence of autologous serum. Mouse macrophages ingested significantly more native as well as oxRBCs in the presence of heat-inactivated fetal calf serum than in the presence of heat-inactivated mouse serum. Pretreated oxRBCs with normal serum were rarely ingested by macrophages in a serum-free medium. Phagocytosis of oxRBCs was significantly inhibited by depletion of IgG or calcium from serum, by heat inactivation of complement, or by antiserum against mouse C3. These results demonstrate that serum components such as IgG, C3, and calcium are involved in phagocytosis of oxRBCs by autologous macrophages.     

3H-Diazepam binding sites in Roman high- and low-avoidance rats

The binding of 3H-diazepam to membrane benzodiazepine receptors was examined in 2 psychogenetically selected lines of rats, which differ according to the selection criterion in avoidance behaviour (RHA/Verh greater than RLA/Verh) and, in addition, in emotionality (RHA/Verh less than RLA/Verh). RHA/Verh rats tended to show higher specific diazepam binding in all CNS-subregions when compared with RLA/Verh animals. Significant differences were found in the cortex, striatum, hippocampus, thalamic region and pons-medulla. These results reinforce the contention that a system involving benzodiazepine receptors may play a role in emotional behaviour.     

Behavior and catecholamine levels: comparative study of gregarious insects, solitary insects, and those treated with carbon dioxide in Locusta migratoria (Insecta, Orthoptera)]

The dopamine (DA) and noradrenaline (NA) was measured in Locusta migratoria for 3 groups of individuals showing differences in their motility: gregarious are very active, solitary and animals in chronic treatment by CO2 (1 mn/day) show a very low motility. NA is present in small amounts (0,120-0,250 microng/g) in the 3 groups without significant differences. On the contrary, the quantity of DA is 5 times greater in gregarious than in the 2 other groups (1,78 and 0,31-0,39 microng/g). Thus it is suggested that DA is related to motility and must play its own role of a neurochormone, distinct from that of a metabolic intermediary between DOPA and NA. The question of a relationship between the metabolism of catecholamine and melanization is open.     

Morphological and biochemical effects of excessive amounts of biotin on embryonic development in mice

Pregnant mice received excessive amounts of biotin either subcutaneously (sc) or orally during gestation. There were no differences in the successful pregnancy rates and number of dead or resorbed fetuses between the control and biotin-treated groups. In biotin-treated groups no increased incidence of fetuses with external malformations was clearly demonstrable. However, biotin accumulated in maternal and embryonic organs; especially, the serum biotin level in the biotin-treated dam was 200-fold higher than that in the control dam. There was a difference in biotinidase activity in maternal serum and placenta between the control and biotin-treated groups. It was concluded that excessive amounts of biotin affected the specific activity of biotinidase in pregnant mice, but did not disturb normal reproductive functions and embryonic development.     

交流电场增强的牛副结核快速血清免疫检测

人/动物发病初期的免疫检测在疾病诊断和防治领域具有重要意义.发病初期病患血清中抗体浓度较低,而传统检测方法单纯依靠抗体自身的随机运动使之与抗原结合发生特异性免疫反应,需要数小时甚至更长的时间才能识别出阳性血清,不利于疾病的快速诊断.因此加快对血清中低浓度抗体的检测速度是提高临床诊断效率的关键因素.本文提出基于交流电热和介电泳技术的血清中低浓度抗体快速检测的新方法,搭建"硅基底非对称平行电极阵列-PDMS微通道"微流控测试平台,以牛副结核为例,通过对免疫反应过程中"电极/血清"界面双电层电容的实时测量,以单位时间内电容的相对变化率为指标,成功分辨出阴性和阳性血清,检测时间仅为2 min.结合交流电场下微流体的流动和可极化粒子的介电泳理论,分析了免疫反应加快的机理.交流电场加速了微通道内抗体分子的对流和传质,大幅度提高了免疫反应效率,结合免疫反应方程,给出了免疫检测过程中微通道内抗体浓度变化的数值仿真,在理论上证明了快速血清免疫检测新方法的可行性.     

Polymorphisme des lipoprotéines et des glycoprotéines sériques chez la Truite

Summary It is shown that there are important differences among the serum proteins of 3 different species of trout. We stress that these genetic variations concern the lipoproteins as well as the glycoproteins. Furthermore, 3 serum proteins ofEsox lucius have antigenic determinants common with the serum proteins ofS. gairdneri; they are an ₂-lipoprotein, an ₂-glycoprotein and a -globulin.