Immunohistochemistry of a spontaneous murine ovarian teratoma with neuroepithelial differentiation. Neuron-associated beta-tubulin as a marker for primitive neuroepithelium

Spontaneous ovarian teratomas develop in a large proportion of female LT strain mice. These tumors display a large neuroectodermal component with morphologic differentiation ranging from primitive neuroepithelium (medulloepithelial and ependymoblastic rosettes) to mature neurons, and provide a useful system for the study of various asynchronous stages of neuroepithelial differentiation. The aim of this study was to assess the expression of various cytoskeletal proteins in conjunction with other differentiation-related antigens in these tumors. We found that the medulloepithelial rosettes reacted with only two anti-beta-tubulin monoclonal antibodies. One of these (TU27) recognizes an epitope common to all of the mammalian beta-tubulin isotypes. The other monoclonal antibody (TUJ1) recognizes an epitope unique to class III beta-tubulin isotypes (neuronal-associated). Whereas immunoreactivity in the ependymoblastic rosettes was limited to TU27, differentiating polar neuroblasts reacted with both TU27 and TUJ1 and expressed neuron-specific enolase, synaptophysin, and the 68 kilodalton subunit of neurofilament protein. In well-differentiated foci, mature neurons were positive for all three neurofilament protein subunits (68, 168 and 200 kilodaltons), microtubule-associated-protein 2, synaptophysin, and neuron-specific-enolase, and reacted with both TU27 and TUJ1. By contrast, glial elements expressed glial fibrillary acidic and S-100 proteins, Leu-7 and TU27 but not TUJ1. Myelin basic protein and myelin-associated glycoprotein reactivity was found in the neuropile of these mature areas. The neuroepithelial components were negative for retinal S-antigen and cytokeratin. The expression of the class III beta-tubulin isotype by medulloepithelial rosettes suggests that this isotype may be one of the earliest markers to signal neuronal commitment in primitive neuroepithelium.     

The pathogenesis of lead encephalopathy

Summary The toxic effects of inorganic lead feedings on the immature brain were studied in the rat pup. Beginning when litters were two weeks old, PbCO₃ was fed to nursing mothers and then to pups directly after weaning. Results in lead-fed pups were compared to age-matched controls and to lead-fed young adult males (60 days old). Anaemia and growth failure developed in both pups and adults. In the second week, more than half the pups developed an encephalopathy, with haemorrhage and oedema predominately in the cerebellum and lead-containing densities in the cerebellar molecular layer. The latter were confirmed by X-ray microanalysis. No lead-fed adults showed signs of an encephalopathy.Cerebellar mitochondria from lead-fed pups, studied polarographically, showed a very early loss of respiratory control and a subsequent inhibition of phosphorylation-coupled respiration with NAD-linked substrates but not with succinate. Compared to the pup cerebellum, these changes were much less marked in immature cerebral mitochondria and were not found in adult cerebral or cerebellar mitochondria.Cerebral and cerebellar homogenates from immature and mature lead-fed animals showed large increases in lead content measured by atomic absorption spectrophotometry. Immature cerebellar mitochondrial lead contents were increased to the same extent as in the homogenates. Mitochondria from immature cerebrum and from both regions in the mature brain showed less immediate and smaller increases in lead content.In conclusion, altered mitochondrial respiration occurs early in regional and age-dependent association with lead encephalopathy in the rat pup. The development of lead encephalopathy also is associated with increased mitochondrial lead concentrations.     

PGD in 47,XXY Klinefelter's syndrome patients

The use of ICSI has been a major breakthrough in the treatment of male infertility. Even azoospermic patients with focal spermatogenesis in the testis, may benefit from the ICSI technique in order to father a child. As ICSI use has become more common, centres have introduced infertility treatment for Klinefelter patients. To date, 34 healthy children have been born using ICSI without PGD, and the conception of one 47,XXY fetus has been reported. In view of the possible risk of an increased gonosome number in the spermatozoa of Klinefelter patients, a safer approach--offering these couples ICSI combined with PGD--has been used, and has resulted in the birth of three healthy children. Couples in which the male suffered from Klinefelter's syndrome were first treated in 1995; these patients were offered ICSI + PGD using FISH technology, notably to enumerate the X and Y chromosomes. ICSI + PGD was performed in 32 cycles of 20 couples with spermatozoa originating from a fresh ejaculate (n = 1), testicular biopsy (n = 21) or frozen-thawed testicular biopsy (n = 10). Normal fertilization occurred in 56.0 +/- 22.4% of the successfully injected oocytes. On day 3 of development, 119 embryos from 29 cycles were of sufficient quality to undergo biopsy and subsequent PGD; a positive result was obtained in 113 embryos. Embryos were available for transfer in 26 cycles, with a mean of 1.6 +/- 0.6 embryos per transfer. Eight pregnancies were obtained, and five resulted in a delivery. A total of 113 embryos from couples with Klinefelter's syndrome was compared with 578 embryos from control couples with X-linked disease where PGD was used to determine gender. A significant fall occurred in the rate of normal embryos for couples with Klinefelter's syndrome (54.0%) compared with controls (77.2%). Moreover, a significantly increased risk of abnormalities was observed for sex chromosomes and autosomes; for each autosome separately, this reached significance level for chromosomes 18 and 21 only. Hence, a cautious approach is warranted in advising couples with non-mosaic Klinefelter's syndrome. Moreover, the use of ICSI + PGD or prenatal diagnosis should be carefully considered.     

Preventive treatment of severe allergic complications caused by Hymenoptera venom. Our experience apropos of 250 cases

Out of a total of 350 referrals for allergy to hymenoptera venom over four years, 250 patients who had one or more severe systemic reactions were selected for a "rush course" of immunotherapy after confirmation of the responsible reagin (skin tests, specific IgE). The method used in the 245 patients who accepted the protocol enabled us to arrive at a maintenance dose of 100 micrograms of venom in three days. Tolerance was excellent; no serious side effects were observed; the course of immunotherapy did not have to be stopped in any of the patients. Clinical efficacy appeared to be satisfactory as no systemic reaction occurred in a fifth (50) of the patients treated who were bitten again, either spontaneously (24 cases), sometimes successfully and repeatedly (11 cases), or intentionally (26 cases). The IgG antivenom antibody was constantly raised from the first month and remained at a stable value. This harmless and effective method should be offered to patients allergic to hymenoptera venom.     

Optimization of a formulation for direct compression using a simplex lattice design

In this paper it is demonstrated how the optimum composition of a mixture for direct compression consisting of -lactose monohydrate, roller-dried -lactose and microcrystalline cellulose can be found using a systematic optimization technique. The experiments were chosen according to a simplex lattice design. The results of these experiments were used to fit a mathematical model, which then can predict the properties of all possible mixture compositions and enables a graphic representation of these properties in the form of contour plots. At a level of 4% the effect of three disintegrants (sodium starch glycolate, croscarmellose sodium and crospovidone) on the properties of the tablets compressed from these filler-binders, was evaluated by superimposing the contour plots of the different tablet responses. It was found that all the disintegrants used were effective in this combination of filler-binders. In order to evaluate drug dissolution rate an extra experiment with crospovidone as the disintegrant was performed, in which oxazepam was used as a test drug.     

6-Hydroxydopamine lesion of the dopamine mesocorticolimbic cell bodies increases (+)-amphetamine self-administration

The effect of 6-OHDA lesions of the dopaminergic mesocorticolimbic cell bodies on intravenous (+)-amphetamine self-administration in the rat was assessed. An acquisition paradigm was used in which the rat had to discriminate between an active and an inactive lever. Each press on the active lever delivered 7.5 g/kg (+)-amphetamine. The lesioned animals acquired this discrimination faster and hence self-administered a larger amount of drug. Thus dysfunction of dopaminergic neurons can induce an enhanced vulnerability to drugs which may be abused by humans.     

Treatment of malignant gliomas in adults with BCNU plus metronidazole

Summary Twenty-six adult patients with astrocytomas were treated with BCNU (1,3-bis(2-chloroethyl)-1-nitrosourea) 180–240 mg/m² 1.V. every 6–9 weeks, with metronidazole 1.5 g/m² p. o. 12 h and 1 h before BCNU and again 6 h and 24 h after BCNU. Of twenty-two evaluable patients, 9 (41%) responded with evidence of reduced tumor size on CT scan, 3 (14%) stabilized and 10 (45%) failed. Patients with no prior chemotherapy or radiotherapy, good performance. status, low grade tumors, and age 50 years had the highest response rates, although differences were not statistically significant. Median survival and duration of response have not been reached with a median follow-up time of ten months. Hematological toxicity was dose-limiting and was probably not augmented by the metronidazole. There was one death from infection that was possibly drug-related. Gastrointestinal toxicity was substantial, and was probably increased by the metronidazole.While the combination of BCNU and metronidazole were tolerable, the response rate seen was no higher than that noted for BCNU alone, and further studies using this dose-schedule are not recommended in astrocytomas.Presented at the 13th International Congress of Chemotherapy, Vienna Austria, August 1983.