[Hepatic resection in the fetal rabbit. Histologic comparison of tissue regeneration in the fetus versus the adult]. / Resezione epatica fetale nel coniglio. Confronto istologico della rigenerazione tissutale del feto e dell'adulto.

Fetal tissues present peculiar features of repair after injury. Although the development of fetal hepatocytes have already been studied in vitro and in transplant models, an in vivo study of fetal liver regeneration is still missed in the literature, to the best of our knowledge. Eight time-dated pregnant California rabbits (23, 24, 25, 30 days of gestational age) and 2 adult male California rabbits were anesthetized following a standardized i.v. protocol (ketamine 50 mg/kg; xilazine 5 mg/kg; propiopromazine 0.75 mg/kg; spontaneous breathing; no anesthetic gas). All the pregnant does underwent a midline laparotomy and a minimal hysterotomy to approach a fetus per each animal. In 2 cases, 1 fetus was delivered and prior to sacrifice the fetal liver was sampled in toto (30 days of gestational age). These pregnancies were allowed to continue to term and were uneventful with a full-term spontaneous delivery of the remaining fetuses. In the other 6 pregnancies, after the hysterotomy, the fetal abdomen was entered through a right-sided longitudinal incision and the liver was partially resected by thermocauterization. Fetal abdomen was closed in 1 layer (non absorbable suture 7-0). The fetus was then returned in the uterus and, after amniotic fluid restoration with warmed saline, the hysterotomy was sutured in double layer (polyglycolic 5-0). Maternal abdomen was closed in 1 layer (polyglycolic 4-0) and the skin in a continuous overlying fashion (silk 3-0). The abdominal cavity of the 2 adult male rabbits was entered through a right subcostal incision. Partial liver resection was performed, and abdominal and skin closure followed the same techniques used for the pregnant does. The treated livers were then sampled in toto at 24, 48, 72 hrs and 4 days after surgery from the fetuses, and at 7 days from the adult rabbits. Histological stains were: H & E; Van Gieson; Masson; Alcian Bleu; PAS. Fetal histology showed a low inflammatory reaction poor in PMN cells with minimal deposition of collagen and a high amount of glycogen in the hepatocytes. The inflammatory response to resection was much more evident in the adult samples as much as the abundant intra and extra-cellular deposition of collagen associated to a minor amount of intracellular glycogen. The peculiar features of liver regeneration in the fetus, deserve further experimental studies.

Lymphocyte traffic changes induced by monolateral vagal denervation in mouse thymus and peripheral lymphoid organs.

In this report we show that after monolateral vagal denervation (vagotomy), performed at the cervical level, a transient effect, lasting about 24h, was produced on lymphocyte release from mouse thymus to peripheral lymphoid organs (spleen and lymph nodes). Labelling thymocytes in situ with fluorescein isothiocyanate (FITC) we note that the export of immature cells, CD4+CD8+, double positive (DP), and double negative, CD4-CD8- (DN), from the thymus was consistently increased 24 and 48 h after vagotomy. Double staining with anti-L3T4 (CD4) and anti-mouse CD8alpha showed that the number of DP and DN cells was significantly higher in both spleen and lymph nodes of vagotomized mice compared to controls (sham-operated), whereas the percentage of CD4+CD8- and CD8+CD4-, single positives (SP), was decreased. Considering thymic cellularity and apoptotic values, we exclude the non-specific effect of stress and suggest that this phenomenon could be in part due to a transient lack of the facilitating influence exerted by vagal efferent fibers on lymphocyte traffic at the cortico-medullary junction of the thymic gland, where mature cells, SP, leave the thymus to enter systemic circulation.

Vagal control of lymphocyte release from rat thymus.

The effects of the efferent vagal innervation on the thymus gland of the rat were investigated in vivo by means of section and stimulation experiments. It has been shown that section of the right vagus produces a decrease in the number of lymphocytes released from the thymus into the venous circulation, an effect that disappears after section of the recurrent laryngeal nerve. Vagal stimulation produces a transient increase in the number of lymphocytes released from the thymus, an effect that also disappears after section of the recurrent nerve. The effects of vagotomy are mimicked by nicotinic blocking agents, which also suppress the effects of vagal stimulation, whereas muscarinic blocking agents were ineffective. It is concluded that the vagal fibres running in the recurrent laryngeal nerve exert a tonic and phasic facilitatory influence on the mechanisms responsible for lymphocyte release from the thymus, and that this action is mediated through nicotinic receptors.

PTH induces modification of transductive events in otosclerotic bone cell cultures.

We studied the effect of PTH (10-100 nM) on transductive mechanisms (adenylate cyclase activity, Ca2+ metabolism, IP3 levels) in cell cultures derived from normal and otosclerotic human bone fragments. The cultured cells were osteoblast-like but with calcitonin-receptors still present and with PTH receptors coupled with the adenylate cyclase system. The results showed that PTH activated adenylate cyclase and increased the intracellular Ca2+ levels with qualitative and quantitative differences between the two cellular populations. In particular, otosclerotic cells responded less to hormone stimulation, which is in accord with the current hypothesis of a desensitization of the receptor/enzyme complex associated with the pathological status.

The S-100 protein causes an increase of intracellular calcium and death of PC12 cells.

The S-100 protein-PC12 cell interaction has been studied as a model system of the possible physiological role played by S-100 protein in the nervous system. The data reported demonstrate that S-100 exerts a cytotoxic action which eventually leads to PC12 cell death, regardless of the cell cycle phase. The effect is specific for the S-100 isoforms, which are made up of two identical subunits and is abolished by a monoclonal antibody directed against the same isoforms. Other isoforms and/or calcium-binding proteins, such as troponin or calmodulin, do not induce the same effects. The action of S-100 on cell viability is not detectable in other cell lines of different embryological origin, such as 3T3, L1210, GH3. S-100 causes a rapid and considerable increase (two- to three-fold) of intracellular Ca2+ concentration in PC12 cells accompanied by cytostatic and cytotoxic action. It is postulated that this action also occurs in vivo, as part of the physiological action of this protein.

Alteration of membrane transductive mechanisms induced by ethanol in human lymphocyte cultures.

Ethanol, in millimolar concentrations, significantly modifies different transductive systems in human lymphocyte cultures. In particular, the presence of alcohol in the medium more than doubles the [Ca2+]i (from 70-90 to 200-250 nM), increasing Ca2+ fluxes from outside, and inhibits the active transport carried out by the calcium pump. The Ca2+ release from intracellular stores is not involved because 10 mM EGTA in the medium completely abolished the rise of [Ca2+]i. Since IP3 levels and cAMP concentrations are also involved in ethanol events (although with opposite effects), it seems that the alcohol may have a specific target on cell membranes (G-proteins) which influence many transductive pathways.

Interleukin-1 alpha: regulation of cellular proliferation and collagen synthesis in cultured human osteoblast-like cells.

In this work the authors studied the effects of interleukin-1 alpha on metabolic activities of human osteoblast-like cells in vitro. The bone nature of the cells was established by assaying for specific bone protein, the osteonectin, and the parathormone receptor, an osteoblast marker. Administration of interleukin-1 alpha to cultured osteoblasts produce an increase in cellular proliferation as suggested by 3H-thymidine incorporation and cell growth studies. Interleukin-1 alpha also affected collagen synthesis confirming its potential role on bone-formation and resorption processes.