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Biomedical subjects

L De Ridder

Publications and source records attributed to L De Ridder.

At least 19 recordsLinked to original sources

Study of dose-rate and split-dose effects on the in vitro micronucleus yield in human lymphocytes exposed to X-rays.

This paper reports the effects of changes in dose-rate and dose-fractionation on the micronucleus (MN) yield in human lymphocytes exposed to 250 kV X-rays. For the investigation of dose-rate effects whole blood samples of four healthy donors were irradiated with doses ranging from 1 to 4 Gy given at various dose-rates between 0.2 and 40 Gy/h. For the higher doses (3 and 4 Gy) a decline in the MN yield became apparent when the dose-rate was reduced below 1.6 Gy/h. This effect was enhanced systematically by a further lowering of the dose-rate. For lower doses (1 and 2 Gy) the reduction in the MN yield was less pronounced: only a small effect was observed for two donors when a dose of 2 Gy was administered at a dose-rate of 0.2 Gy/h. In the split-dose experiment a dose of 4 Gy was delivered either as a single exposure or in two fractions of 2 Gy, separated by time intervals ranging from 30 min to 10 h. A continuous decrease of the MN yield with increasing interfraction time is observed: after an initial fast decline a further slight reduction in the MN yield occurs. The observed dose-rate and split-dose effects on the MN yield can be attributed to repair of sublethal damage.

Chromosome Aberrations

Nimodipine decreases resuscitability in a cardiopulmonary arrest model in the rat.

Although calcium has been implicated in ischemia-induced brain death or dysfunction, many animal studies do not show a beneficial effect of calcium-entry blockers given after resuscitation from a cardiopulmonary arrest (CPA). This may be due to the fact that treatment was started too late; we, therefore, evaluated the effect of the calcium-entry blocker nimodipine administered at the earliest feasible postischemic moment, i.e. at the start of the resuscitation attempts. In anesthetized Wistar rats, CPA was induced by an intra-cardiac injection of KCl, and maintained for 7 min by chest restriction. At the start of the resuscitation attempts, 50 rats were blindly and randomly assigned to intravenous treatment with either nimodipine (10 micrograms/kg over 2 min, followed by 1 micrograms/kg per min for 60 min; n = 25) or saline (n = 25). In the nimodipine group, significantly less rats could be resuscitated (11/25 versus 20/25) and the survival rate at the end of the 7 days evaluation period tended to be lower (5/25 versus 11/25). In the rats surviving after 7 days, there was no difference between both groups in incidence of seizures, neurological status and histological lesions in the hippocampus. It is concluded that nimodipine, in the dose tested and given during resuscitation in this rat model, has a detrimental effect on resuscitability and no beneficial effect on the neurological outcome in the surviving animals.

Animals

The healing of cranial defects by demineralized osseous implants: a radiographic, histological and radioisotope-uptake study in rats.

The purpose of this study was to evaluate by different techniques the healing effect of demineralized allogeneic bone in parietal cranial defects in rats. 45Ca incorporation was measured for nine consecutive weeks in groups of control and implanted rats. Repeated 99mTc MDP uptake measurements were performed for nine weeks on the same group of rats. Each measurement was preceded by a cranial radiography. At the end of the study the animals were sacrificed for histologic evaluation. The results of the 45Ca incorporation, 99mTc MDP uptake measurements and the histologic pictures confirmed the osteogenic capacity of demineralized allogenic bone. From the results it can concluded that 99mTc MDP uptake measurements are a powerful tool in the evaluation of the healing process. It can be utilized sequentially in the same animal and gives detailed information about the progress of osteogenic activity over the entire implant area.

Animals

Nimodipine has no beneficial effect on neurological outcome in a cardiopulmonary arrest model in the rat.

Brain damage after resuscitation from cardiac arrest is believed to be related to calcium influx in ischaemic neurons and to postischaemic calcium-dependent vasospasm. We therefore evaluated the potentially protective effects of the calcium-entry blocker nimodipine in a cardiopulmonary arrest model in the rat. Male Wistar rats were anaesthetized with ketamine (group I) or hexobarbital (group II) and subjected to a KCl-induced cardiac arrest during 7 min (group I) or 12 min (group II). Five minutes after resuscitation, the rats were treated intravenously in a randomized and blind fashion. Group I received either saline or 1 microgram.kg-1.min-1 or 5 micrograms.kg-1.min-1 of nimodipine and group II either saline or 1 microgram.kg-1.min-1 of nimodipine. Survival, occurrence of seizures and neurological status were assessed daily during 7 days after resuscitation. On day 7, the brains of the surviving rats were perfusion-fixed and a histopathological evaluation of the hippocampus was performed. Nimodipine, in the doses tested, had no beneficial influence on the 7 day survival rate, nor on the occurrence of seizures and the neurological and histopathological scores in the rats surviving after 7 days. With the highest dose of nimodipine, there was even a trend towards a decrease of the survival rate, probably related to the drug's hypotensive effect. Therefore, our data do not show a protective effect of nimodipine after cardiac arrest.

Animals

Development of larval Echinococcus granulosus confronted with chicken heart tissue in vitro.

Vesiculated protoscoleces (VP) were produced by culturing freshly collected protoscoleces from Echinococcus granulosus horse liver hydatids in RPMI 1640 monophasic medium at 37 degrees C for 18 days. Half of the VP were used as such, the other half used after killing them by freeze-thawing. Nine-day-old chicken heart fragments (CHF) were cultured in MEM at 37 degrees C for 72 h. Subsequently, CHF were put together with live and dead VP, respectively, for up to 53 days, on a semisolid medium consisting of agar, Ringer's and MEM. Time-dependent histological observations revealed that dead VP were surrounded by CHF cells. Dead VP tissue was eventually internalized and disintegrated in about 1 week. Live VP penetrated into the CHF tissue and further developed into small hydatid cysts, located within the boundaries of the experimental 'host' tissue. The amorphous-looking contact region PAP-stained positively only with anti-E. granulosus serum and not with anti-CHF serum; it was considered identical to the normal laminated layer. The invasion of VP in CHF tissue proved to be different from a tumour or a bacterial invasion: it was concluded that the confrontation of VP and CHF had resulted in an 'in vitro cohabitation' rather than in an 'in vitro infection'.

Animals

Dose- and time-dependent radiation inhibition of RNA and glycosaminoglycan synthesis in embryonic cartilage: an in vitro study.

Radiation effects on the RNA and glycosaminoglycan (GAG) synthesis of embryonic cartilaginous tibiae were studied in vitro during a 4- or 7-day culture period. Before being cultured, tibiae received single radiation doses of 20, 50 or 100 Gy. The counterparts served as sham-irradiated controls. At different times after irradiation, irradiated and control tibiae were pulse-labelled for 2 h with [3H]uridine or [3H]glucosamine. The incorporated radioactivity was measured by liquid scintillation counting. Histochemical demonstration of acid phosphatase (AP), a lysosomal enzyme, was carried out using beta-glycerophosphate as substrate. A limited, dose-dependent immediate effect on RNA and GAG synthesis was found. This effect was unchanged for 2 days. After this period a time-dependent delayed effect was observed. For each radiation dose, and for each precursor, the same time-related pattern was found. At the end of the culture period AP activity, an early indicator for apoptosis, was higher in the irradiated tibiae than in the controls. No other morphological ultrastructural differences were observed at this time. We conclude that the metabolic alterations are probably due to stimulation of the initial stages of the apoptotic process in the irradiated cartilage cells.

Animals

Invasiveness, proliferative activity and ultrastructural phenotypes of hepatocytes from diethylnitrosamine-induced neoplastic nodules and hepacarcinomas in vitro.

The invasive behavior of hepatocytes from diethylnitrosamine (DENA) induced neoplastic nodules and hepatocarcinomas was studied in a confronting culture system in vitro. These observations confirm our previous report, demonstrating that hepatocytes from hepatocarcinomas and from neoplastic nodules invaded into embryonic chick precultured heart fragments (PHF), a property associated with malignancy (Mareel, 1979). We now further demonstrate that: (1) Invasiveness was expressed by the hepatocytes in 10 out of 12 samples from hepatocarcinomas, and in 13 out of 36 confronted nodular samples. The hepatocytes from the other two-thirds of nodule samples died off in the confrontation as did all normal hepatocytes. (2) Invasive hepatocytes from tumors and from nodules showed the same arrangement of invasive liver cells in relation to the heart tissue, and the same ultrastructural phenotypes. The latter did not differ from those in the non-invading subpopulations, with the exception perhaps of a higher proportion of cells with an indented nucleus. (3) None of the scored ultrastructural alterations was present in all the invasive cells, thus excluding any specific requisite in this respect. (4) When 3H-TdR was made continuously available to the cultures, starting at the time of confrontation between heart and liver tissues, unlabelled as well as labelled invasive hepatocytes were found inside the PHF in about equal proportions. It is concluded that nodular hepatocytes deviate from normality by at least 2 different properties that may or may not be related to each other and are revealed under in vitro conditions, namely the ability to survive and to proceed through S-phase and mitosis under such conditions and to actively invade precultured chick heart fragments. The latter property indicates that at least some of the nodules contain hepatocytes that have performed one step in malignant progression.

Animals

Interaction between embryonic rat superior cervical ganglion and syngeneic heart fragments in a confronting culture.

Freshly dissected 17-19 days embryonic superior cervical ganglia are confronted with 0.5 mm diameter heart fragments of the same Wistar rat. Incubation in vitro at 37 degrees C of confronting heart-ganglion pairs is carried out in Dulbecco's medium on a gyratory shaker. Fixation and staining followed after 2 hr, 1, 3 and 6 days of incubation. Histological analysis with light and electron microscopy revealed the interaction between the sympathetic ganglion and its target organ. Ingrowth of axons rich in neurofilaments and neurotubules and containing light and dense core vesicles, is observed.

Animals

Morphology and 125I-concentration of embryonic chick thyroids cultured in an atmosphere of oxygen.

In vitro culture of whole thyroid glands may lead to loss of differentiation and function. Comparison by histology, histoautoradiography and gammacounting of 125I-concentration, of 12 to 13 days old embryonic chick thyroids, cultured on a gyratory shaker either in an atmosphere of 5% CO2 and air or of 95% O2 and 5% CO2, shows that a high concentration of O2 maintains the morphological and functional integrity of the thyroids for at least 4 days.

Animals

Characterization of spontaneous, chemicak, and viral transformants of a C3H/3T3-type mouse cell line by transplantation into young chick blastoderms.

Cells from a C3H/3T3-type cell line were transplanted into defects of the lower layer of stage-4 chick blastoderms before and after "spontaneous," chemical, and viral transformation. To check the validity of inhibition of lower-layer defect closure as a criterion of malignancy, we compared the behavior of the lower layer toward these cells with their in vitro growth pattern, their capacity to invade embryonic chick skin explants that were organotypicaly cultured, and their tumorigenicity in syngeneic mice. No false-positive results were observed with either test. The study of the in vitro growth pattern gave false-negative results during the early phase of spontaneous transformation, whereas these cells, exhibiting an untransformed growth pattern, were shown to be malignant by the other tests. We concluded that the inhibition of lower-layer defect closure is a reliable, sensitive and rapid test for the detection of malignancy in tissue-cultured cells from any source.

Animals