Alkaline phosphatase isozyme activity in serum from patients with chronic periodontitis.

BACKGROUND: High alkaline phosphatase activity (ALP) is shown in the periodontal ligament due to the constant renewal of this tissue or pathological circumstances. We have previously shown that the activity level of this enzyme could be reflected at the serum level. OBJECTIVES: Because the local production of ALP in the periodontal ligament is often of the bone-type enzyme, we studied the activity of this isozyme among the other isoforms in the serum of patients with chronic periodontitis in comparison with that of control subjects. MATERIAL AND METHODS: This study included 83 patients (59 with periodontal disease, 24 as control group) and we determined the total seric ALP activity and the percentage of the different isoforms (essentially bone, kidney and intestinal-types) by Ektachem analyser and gel agarose electrophoresis respectively. CONCLUSIONS: By comparisons between the two groups, our results showed a relationship between loss of attachment in periodontal disease and a drop in bone ALP activity in serum. Moreover, these results suggested a gender based difference as well, with lower activity more frequent in women than in men.

An activity-dependent neurotrophin-3 autocrine loop regulates the phenotype of developing hippocampal pyramidal neurons before target contact.

Neurotrophin-3 (NT-3), its cognate receptor trkC, and voltage-gated calcium channels are coexpressed by embryonic pyramidal neurons before target contact, but their functions at this stage of development are still unclear. We show here that, in vitro, anti-NT-3 and anti-trkC antibodies blocked the increase, and NT-3 reversed the decrease in the number of calbindin-D(28k)-positive pyramidal neurons induced by, respectively, calcium channel activations and blockades. Similar results were obtained with single-neuron microcultures. In addition, voltage-gated calcium channel inhibition downregulates the extracellular levels of NT-3 in high-density cultures. Moreover, electrophysiological experiments in single-cell cultures reveal a tetrodotoxin-sensitive spontaneous electrical activity allowing voltage-gated calcium channel activation. The mouse NT-3 (-/-) mutation decreases by 40% the number of developing calbindin-D(28k)-positive pyramidal neurons, without affecting neuronal survival, both in vitro and in vivo. Thus, present results strongly support that an activity-dependent autocrine NT-3 loop provides a local, intrinsic mechanism by which, before target contact, hippocampal pyramidal-like neurons may regulate their own differentiation, a role that may be important during early CNS differentiation or after adult target disruption.

Q- and L-type calcium channels control the development of calbindin phenotype in hippocampal pyramidal neurons in vitro.

Cultured immature hippocampal neurons from embryonic 17-day-old rats were used to explore activity-dependent regulation of neuronal phenotype differentiation in the developing hippocampus. The calbindin-D28k phenotype of the pyramidal neurons appeared during the first 6 days in culture, and was expressed by 12% of the cells on day 6. Daily stimulation with 50 mM KCl during the first 5 days in vitro increased the number of calbindin-D28k-positive pyramidal neurons without affecting neuronal survival. This effect was prevented by buffering extracellular Ca2+. Omega-agatoxin-IVA-sensitive Q-type and nitrendipine-sensitive L-type voltage-gated Ca2+ channels (VGCCs) carried Ca2+ currents and Ca2+ influx in immature pyramidal neurons at somata level. Blockade of these channels inhibited calbindin-D28k phenotype induced by 50 mM KCl. Conversely, glutamate-activated Ca2+ channel antagonists did not affect the KCl-induced calbindin-D28k phenotype. Chronic blockade of Q- and/or L-type VGCCs downregulated the normal calbindin-D28k development of immature pyramidal neurons without affecting neuronal survival, the somatic area of pyramidal neurons or the number of GABAergic-positive (gamma-aminobutyric acid) interneurons. However, at later developmental stages, Q-type VGCCs lost their ability to control Ca2+ influx at somata level, and both Q- and L-type VGCCs failed to regulate calbindin-D28k phenotype. These results suggest that Q-type channels, which have been predominantly associated with neurotransmitter release in adult brain, transiently act in synergy with L-type VGCCs to direct early neuronal differentiation of hippocampal pyramidal neurons before the establishment of their synaptic circuits.

[Detection of liver metastases from digestive cancer. Value of alkaline phosphatases, their macromolecular isoenzyme and of ceruloplasmin]. / Détection de métastases hépatiques d'un cancer digestif--intérêt des phosphatases alcalines, de leur isoenzyme macromoléculaire et de la céruléoplasmine.

In order to improve the non aggressive diagnosis of hepatic metastasis from digestive neoplasm, the authors analyzed the following biological parameters: aminotransferases, alkaline phosphatase and lacticodehydrogenase isoenzymes, gammaglutamyl-transpeptidase, conjugated and total bilirubin, C-reactive protein, type A, G, M immunoglobulins, C3 complement factor, alpha-1 acidic glycoprotein (orosomucoid), haptoglobin, ceruloplasmin, transferrin, albumin, prealbumin, ferritin. This work included 54 patients with digestive tract cancer (esophageal, gastric, colic, rectal, anal localizations), divided in two groups: M- (n = 27), without hepatic metastasis), and M+ (n = 27, with histological confirmed hepatic metastasis). The Mann-Whitney test showed significant differences for 12 parameters between the 2 groups. With more than 60% sensitivity (Se) and specificity (Sp), according to the ROC curves, the following parameters can be selected: Total alkaline phosphatase (Se 89%, Sp 70%) and their macromolecular H2 fraction, lacticodehydrogenase fraction 4 (Se 63%, Sp 63%), gammaglutamyl-transpeptidase (Se 85%, Sp 82%), ceruloplasmin (Se 64%, Sp 65%), aspartate-aminotransferase determination (Se 63%, Sp 65%).

Experimental studies on unconjugated bilirubin binding by human erythrocytes.

The evaluation of the bilirubin bound to human erythrocytes is considered by some paediatricians as a test to estimate the risk of development of kernicterus. We have studied the physical and chemical characteristics of this binding. Red blood cell membranes contain specific binding sites for bilirubin, the affinity of which is low (Kd = 170 mumol/L). The dissociation constant of the bilirubin/human serum albumin complex is about 10,000 times lower. In jaundiced neonates even with a level of blood bilirubin higher than 300 mumol/l, the binding of bilirubin to red blood cells is negligible. So, the evaluation of the bilirubin bound to human red blood cells does not seem to be a useful test to appreciate the risk of development of kernicterus.

Study of human isoferritins from liver, spleen, heart and placenta by the microcomplement fixation technique.

Ferritin, iron storage protein, presents two types of subunits, H and L, the respective proportions of which varying with the tissue of origin and defining molecules called isoferritins. This work attempts to compare four human ferritins (from liver, spleen, heart and placenta) by means of the microcomplement fixation technique. Results show that placenta and liver ferritins are closely related, while slight but significant differences appear between liver, spleen and heart ferritins. These differences are obviously less important compared to those observed between ferritins from different species of origin, as shown by the results expressed in terms of index of dissimilarity or immunological distance. Those results are fully consistent with the elementary aminoacid composition as well as with the relative proportions of H and L subunits among the various types of human isoferritins we have studied.