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E Sikora

Publications and source records attributed to E Sikora.

At least 55 records · Page 3Linked to original sources
PubMed1990

Subcortical afferents to the interstitial nucleus of Cajal: an anatomical retrograde tracing study in the rabbit.

Organization of brainstem projections to the interstitial nucleus of Cajal (INC) in the rabbit has been studied using the method of retrograde transport of horseradish peroxidase or wheat germ agglutinin-horseradish peroxidase conjugate. Injections of tracers into INC resulted in bilateral labelling in the medial terminal nucleus of the accessory optic tract, medial region of the zona incerta, vestibular nuclei (superior, medial, inferior), rostral portion of the prepositus nucleus and several nuclei of the pontine and medullary reticular formation. Retrograde labelling on the contralateral side was noted in all 4 deep cerebellar nuclei, the lateral vestibular nucleus, group Y, the rostral interstitial nucleus of the medial longitudinal fascicle, INC and mesencephalic reticular formation dorsal and lateral to the red nucleus. Cells of origin for the ipsilateral afferents of INC were found only in the nucleus of the posterior commissure. These data are discussed in relation to other morphological and physiological studies of afferent connectivity of INC in other species.

Animals↗
PubMed1988

Formation and retention and biological activity of N10-propargyl-5,8-dideazafolic acid (CB3717) polyglutamates in L1210 cells in vitro.

The formation, retention and biological activity of the polyglutamate metabolites of the thymidylate synthase (TS) inhibitor N10-propargyl-5,8-dideazafolic acid (CB3717) has been investigated in L1210 murine leukaemia cells grown in vitro. CB3717 polyglutamates were measured by HPLC using high specific activity 3H-CB3717. Following the exposure of cells to 50 microM CB3717 for 6, 12 and 24 hr total cellular radioactivity corresponded to 4.5 +/- 1.5, 6.8 +/- 3.6 and 5.9 +/- 3.4 microM drug derived material, respectively. Of this material, greater than 70%, 57 +/- 3% and 51 +/- 5% was in the form of unchanged CB3717 at 6, 12 and 24 hr respectively. The remaining radioactivity was associated with polyglutamate metabolites of CB3717, predominantly the tetra and pentaglutamate forms. Following the removal of extracellular drug after incubation for 24 hr and resuspension in drug free medium, unchanged CB3717 was lost rapidly from the cells such that after 6 hr it accounted for only 5% of total cellular radioactivity. In contrast, levels of CB3717 tetra and pentaglutamates declined solely due to dilution during cell division. Measurement of the whole cell TS activity by 3H-deoxyuridine incorporation into DNA indicated that, despite the loss of unchanged CB3717 from the cell, enzyme activity remained suppressed (less than 10% of control) for at least 24 hr after resuspension in drug free medium. The TS inhibitory activity of the polyglutamated metabolites of CB3717 was investigated using enzyme purified from L1210 cells. As inhibitors, the metabolites were 26-, 87-, 119- and 114-fold more potent than CB3717 as the di-, tri-, tetra- and pentaglutamate forms, respectively. However, as inhibitors of dihydrofolate reductase prepared from rat liver, CB3717 polyglutamates were no more than 5-fold More potent than the parent compound. This study has shown that CB3717 can undergo polyglutamation in tumour cells and that the metabolites are preferentially retained giving rise to prolonged TS inhibition. By virtue of their potent TS inhibitory activity these metabolites are, therefore, most probably the intracellular effectors of CB3717 cytotoxicity.

Animals↗
PubMed1988

Development of an assay for the estimation of N10-propargyl-5,8-dideazafolic acid polyglutamates in tumor cells.

A method is described herein for the isolation and quantitation of polyglutamates of the thymidylate synthase (TS) inhibitor N10-propargyl-5,8-dideazafolic acid (CB3717) in tumor cells exposed to the drug in vitro. Cells were incubated with 50 microM 3H-CB3717 for 12 h and then disrupted by sonication. CB3717 and its polyglutamates were extracted by boiling in 0.01 M Tris-HCl pH 10. The extract was concentrated by lyophilization and analyzed by reverse phase HPLC (10 x 0.46-cm Polygosil 5-micron C18 column) using linear gradient elution (5-16% acetonitrile in 0.1 M sodium acetate, pH 5, over 15 min, 2 ml/min). Recovery of radioactivity at each stage of the method was greater than 70%. CB3717 and its polyglutamates were identified by co-chromatography with synthetic standards and by inhibition of partially purified TS. Quantitation was by means of radiochemical analysis. The 3H-CB3717 used in these studies was prepared by catalytic tritiation of diethyl-(2-chloro-4-nitrobenzoyl)-L-glutamate followed by consecutive alkylation with propargyl bromide and 2-amino-6-bromomethyl-3,4-dihydro-4-oxoquinazoline hydrobromide. The free diacid was prepared as required by hydrolysis in sodium hydroxide and purified by HPLC. Tritiation in only one position was confirmed by 3H NMR. Following the exposure of L1210 leukemia cells to 50 microM 3H-CB3717 for 12 h the total cellular radioactivity level was approximately 7 microM, of which 27% was present as polyglutamated metabolites with four and five glutamate residues.

Animals↗
PubMed1987

Recent preclinical and clinical studies with the thymidylate synthase inhibitor N10-propargyl-5,8-dideazafolic acid (CB 3717).

CB 3717, N10-propargyl-5,8-dideazafolic acid, is a tight-binding inhibitor of thymidylate synthase (TS) whose cytotoxicity is mediated solely through the inhibition of this enzyme. Recent preclinical studies have focused on the intracellular formation of CB 3717 polyglutamates. Following a 12-hour exposure of L1210 cells to 50 microM [3H]CB 3717, 30% of the extractable radioactivity could be accounted for as CB 3717 tetra- and pentaglutamate, as determined by high-pressure liquid chromatography (HPLC) analyses. As inhibitors of isolated L1210 TS, CB 3717 di-, tri-, tetra- and pentaglutamate are 26-, 87-, 119- and 114-fold more potent than CB 3717, respectively, and their formation may, therefore, be an important determinant of CB 3717 cytotoxicity. In early clinical studies with CB 3717, activity has been seen in breast cancer, ovarian cancer, hepatoma, and mesothelioma. Toxicities included hepatotoxicity, malaise, and dose-limiting nephrotoxicity. This latter effect is thought to be due to drug precipitation within the renal tubule as a result of the poor solubility of CB 3717 under acidic conditions. In an attempt to overcome this problem, a clinical trial of CB 3717 administered with alkaline diuresis is under way. Preliminary results at 400 and 500 mg/m2 suggest that a reduction in nephrotoxicity may have been achieved with only 1 instance of renal toxicity in 10 patients. Hepatotoxicity and malaise are again the most frequent side effects. Evidence of antitumor activity has been seen in 3 patients. Pharmacokinetic investigations have shown that alkaline diuresis does not alter CB 3717 plasma levels or urinary excretion and that satisfactory urinary alkalinization can be readily achieved.

Adult↗
PubMed1986

The production of focal herpes encephalitis in mice by stereotaxic inoculation of virus. Anatomical and behavioral effects.

A low virulence strain of herpes simplex type 1 was microinjected into the hippocampus of BALB/c mice. Intense replication of virus at the inoculum site was followed by spread of viral antigen to the afferent connections of the hippocampus. Surviving animals showed focal damage of limbic structures and specific behavioral abnormalities generally consistent with hippocampal damage. This procedure thus produces an animal model which more closely resembles human herpes encephalitis than those previously reported.

Animals↗
PubMed1986

Age- and tumour-related changes in methionine biosynthesis in mice.

Two routes for the methylation of homocysteine to methionine, depending either on betaine or folate cofactor as methyl donor, were studied in liver and kidneys of normal and Ehrlich ascites carcinoma-bearing mice at various stages of their postnatal development. Distinct age-dependence in the activities of betaine methyltransferase, methylenetetrahydrofolate reductase and methionine synthase were found both in normal and tumour-bearing mice. Independent of the levels of enzyme activity in healthy mice, the tumour activated one route of methionine formation only, namely that utilizing methyltetrahydrofolate as the methyl donor. This effect was observed in host liver exclusively. No host age-related changes were found in Ehrlich ascites carcinoma growth.

5-Methyltetrahydrofolate-Homocysteine S-Methyltran↗
PubMed1984

Folate enzymes in Ehrlich ascites carcinoma-bearing mice.

The activity of 4 enzymes involved in the formation and interconversion of folate coenzymes has been examined in liver and kidney of healthy and Ehrlich ascites carcinoma-bearing mice. In the liver, a 50% increase of methylenetetrahydrofolate reductase activity was shown soon after tumour cell inoculation, while the activity of formyltetrahydrofolate synthetase and methylenetetrahydrofolate dehydrogenase decrease by 20% at an advanced stage of tumour development. In kidneys of the host mouse the only change observed was a decrease of dihydrofolate reductase activity. The levels of activity of all assayed enzymes found in host organs were similar to that in Ehrlich carcinoma cells.

5,10-Methylenetetrahydrofolate Reductase (FADH2)↗
PubMed1984

Comparison of folylderivative biosynthesis in Ehrlich ascites carcinoma cells and in some organs of healthy and tumor-bearing mice.

Biosynthesis of folylderivatives derived from subcutaneously injected 2-[14C]folate was studied in Ehrlich ascites carcinoma (EAC) cells and in liver and kidneys of healthy and tumor-bearing mice. Retention of exogenous folate was followed by measurements of the total radioactivity of folylderivatives present in the EAC cells and organs examined. Identification of unconjugated and conjugated folylderivatives was done by means of column chromatography on Sephadex G-25, G-15 and cellulose sheets. The level of retained radioactivity in folyderivatives, being 5% in the liver and 1% in the kidneys of the radioactivity administered to mice, was similar in healthy and tumor-bearing animals. Moreover, no quantitative and qualitative differences were found in folylmono-and polyglutamates originated from the organs of healthy or tumor-bearing mice, although the content of folylpolyglutamates arose faster in liver and kidneys of EAC cells-bearing mice as well as in the tumor cells than in the organs of healthy mice.

Animals↗
PubMed1983

Disappearance of scrapie virus from tissues of the mouse.

Examination of newborn mice, inoculated intraperitoneally with high doses of scrapie virus, revealed that the virus could not be reisolated from their tissues after about 1 week following inoculation, until almost 1 year later. The inoculum was rapidly removed and was not detectable, although the animals became latently infected. Homogenization of whole inoculated newborn animals showed that only about 3% of virus could be recovered by the 2nd day postinoculation (p.i.). During the first 6 days p.i. the half-life of titratable scrapie was about 15 h. A further study of the rate of disappearance of clarified scrapie virus from blood after intravenous inoculation showed an even more rapid disappearance, with a half-life of 5.16 min. Prior treatment of the recipient mice with either carbon black or silica to block the reticuloendothelial system (RES) did not affect the rate of disappearance. It was concluded that the mouse possesses a very efficient means of scrapie virus removal from the blood which is not dependent upon an active RES. However, after 1 h the rate of disappearance changed dramatically; the residual virus level was very stable, with no significant drop during the next 21 h. This finding was compatible with the possibility that two forms of scrapie virus, with different removal rates, coexisted in the inoculum. Silica treatment caused a shortened scrapie incubation period.

Age Factors↗
PubMed1983

A quinazoline antifolate as inhibitor of folate metabolism in mouse L-cells.

The effects of quinazoline antifolate CB 3703 on folate metabolism in mouse L-cells were examined by measuring the amount of 14C label in folate derivatives and its distribution in separated folylmono- and polyglutamates in the cells grown in the presence of radioactive folate and quinazoline. The total radioactivity of folyl derivatives in the quinazoline-treated cells did not exceed 15% of the radioactivity extracted from the control cells, in spite of the fact that quinazoline CB 3703 did not interfere with the uptake of folate into L-cells. Analysis of folylmono- and polyglutamate derivatives, either incubated or not incubated with gamma-glutamylcarboxypeptidase, by chromatography on Sephadex G-25, G-15 or DEAE-cellulose revealed that in the quinazoline-treated cells practically all radioactivity was found in the position of unmetabolized folate. It seems probable, therefore, that quinazoline CB 3703 impairs in L-cells folate reduction and conversion to its coenzymatic forms and their subsequent polyglutamation. These facts seem to be responsible for a decrease in retention of folate derivatives in the quinazoline CB 3703-treated cells.

Animals↗
PubMed1980

Locomotor effects of catecholaminergic drugs on herpes-infected mice.

Changes in spontaneous, amphetamine (AMP) and apomorphine (APO) induced locomotor activity were used to assess the effects of central nervous system (CNS) infection with herpes type 1 virus. A dual herpesvirus inoculation procedure was used in which the animals received an immunizing footpad inoculation followed at 2 weeks by an identical intracerebral challenge. Four weeks later the animals were tested with intraperitoneal injections of saline or d-l-amphatmine (0.5 and 2.0 mg/kg). When footpad herpes-virus was given via one or two injections, it had no effect on spontaneous or AMP induced activity. When food-pad-intracerebral herpes mice were tested 28-33 days post intracerebral inoculation, they demonstrated depressed AMP-induced but not spontaneous activity. AMP at a dosage of 5.0 mg/kg overcame the herpesvirus blockage of 0.5 and 2.0 mg/kg AMP induced activity. Intraperitoneal injection of APO in day 3 post-IC animals produced less suppression of activity in the virus group than in the controls. These results suggest that non-fatal CNS herpes infection produces hypoactivity, in contrast to thehyperactivity during acute fatal CNS herpes encephalitis (Lycke & Roos, 1975), and that the effect may be due to alterations in postsynaptic receptor sensitivity.

Amphetamine↗
PubMed1976

Blast crisis of chronic myeloid leukaemia: the effect of intensive chemotherapy.

24 patients with Philadelphia chromosome positive chronic myeloid leukaemia (CML) in blast crisis were treated with intensive chemotherapy. 16 patients showed either partial or complete response to this treatment, but median survival remained short (13 weeks), and much of this time was spent in hospital. These results were not significantly better than those obtained by others using vincristine and prednisolone alone, and this combination of drugs can often be given on an outpatient basis. It is concluded that until more effective intensive therapy becomes available patients in CML blast crisis should be managed in such a way that the quality of life is not impaired; and that at present vincristine and prednisolone appears to be the most impaired; and that at present vincristine and prednisolone appears to be the most appropriate initial treatment, though even this is far from satisfactory.

Adolescent↗