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J Piñero

Publications and source records attributed to J Piñero.

At least 37 records · Page 2Linked to original sources
PubMed1994

Differences in the adaptive response to radiation damage in G0 human lymphocytes conditioned with hydrogen peroxide or low-dose X-rays.

We have carried out experiments to study the adaptive response in G0 human lymphocytes conditioned with either hydrogen peroxide or low-dose X-rays and challenged with 1.5 Gy of X-rays after stimulation. Peroxide conditioning treatment was given at different times before stimulation, while the low-dose irradiation was delivered at different dose rates just before stimulation of lymphocytes. A protective effect of pre-exposure to H2O2 against radiation damage detected as micronuclei in binucleated cells was evident, regardless of the time of conditioning treatment during G0. For low-dose-irradiated cells, on the other hand, the adaptation observed seemed to depend upon the dose rate, and never reached the extent observed in cells treated with peroxide.

Adaptation, Physiological↗
PubMed1994

Evidence that SCEs induced by mutagens do not occur at the same locus in successive cell cycles: lack of cancellation in three-way stained CHO chromosomes.

An approach based on the synchronization of CHO cells after a first cell cycle incorporating a relatively low amount of bromodeoxyuridine (BrdUrd) into DNA, followed by mutagenic treatment and subsequent culture for second and third generations of BrdUrd incorporation for the scoring of sister chromatid exchanges (SCEs) per cell cycle in three-way differentially (TWD) stained chromosomes, has been used to investigate the possible cancellation of SCEs. Cancellation is expected to occur if two mutagen-induced SCEs occur at exactly the same site in subsequent rounds of replication. Lesions in DNA seem to persist and are able to induce SCE throughout two cell cycles after treatment with the three mutagens tested--mitomycin C (MMC), ethyl methanesulfonate (EMS) and ultraviolet (UV) light--though this latter agent was shown as only moderately persistent. Our results seem to indicate that SCEs induced by these mutagens do not take place at the same locus in successive cell generations, as assessed by a lack of SCE cancellation.

Animals↗
PubMed1994

Synergistic effect of inhibitors of topoisomerase I and II on chromosome damage and cell killing in cultured Chinese hamster ovary cells.

Simultaneous treatment of cultured Chinese hamster ovary cells with the topoisomerase I inhibitor camptothecin and the topoisomerase II inhibitor 4'-(9-acridinylamino)-methanesulfon-m-anisidide results in a clear synergistic effect on both chromosome damage detected at metaphase and loss of colony-forming ability. In contrast, the effect of combined treatment with these topoisomerase inhibitors on sister chromatid exchanges was not significantly different from that expected if the effects were additive. Taken as a whole, these results seem to support the hypothesis that topoisomerase inhibitors can lead to cell death, presumably when DNA replication forks collide with drug-stabilized cleavable complexes. Nevertheless, no evidence of apoptosis was obtained from DNA fragmentation analysis. The possible clinical implications of our findings are discussed.

Amsacrine↗
PubMed1993

Cytogenetic effects of inhibition of topoisomerase I or II activities in the CHO mutant EM9 and its parental line AA8.

The possible relationship between topoisomerase activities and the occurrence of SCE and chromosomal aberrations was investigated in the CHO ethyl methanesulfonate-sensitive mutant EM9 and its parental line, AA8. The Topo II inhibitor m-AMSA induced fewer SCE in EM9 than in AA8, mainly when given during the first S period. When the Topo I inhibitor camptothecin was used, it showed a higher efficiency to induce both chromosomal aberrations and SCE in EM9 than in AA8. The impact of BrdU incorporated into parental DNA on topoisomerase activity was tested using nuclear extracts from both EM9 and AA8 assayed for their ability to decatenate kinetoplast DNA by Topo II and to relax supercoiled plasmid DNA by Topo I. Taken as a whole, the results seem to indicate that there are differences between the two cell lines, consistent with the hypothesis put forward by other investigators that topoisomerases are involved in the molecular mechanism of SCE.

Amsacrine↗
PubMed1993

Abnormally high incidence of SCE in three successive cell cycles in the CHO mutant EM9 as detected by a three-way immunoperoxidase differential staining.

A high resolution three-way immunoperoxidase method for the detection of SCE per cell generation in chromosomes with very low levels of BrdU in DNA has been recently standardized in our laboratory. We have made use of this methodology to assess whether the CHO mutant EM9 is solely hypersensitive to BrdU incorporated into template DNA as regards the extraordinarily high SCE frequency observed as compared with its parental line, AA8, or also shows an elevated spontaneous frequency of exchanges. Our results confirm the importance of BrdU incorporated for SCE in EM9 but also show that the estimated yield of spontaneous SCE is about 10 times higher in this mutant than in the parental line, AA8.

Animals↗
PubMed1993

Importance of replication fork progression for the induction of chromosome damage and SCE by inhibitors of DNA topoisomerases.

Previous studies have shown the importance of DNA replication fork progression for the cytotoxicity of topoisomerase inhibitors. Nevertheless, while it was concluded that an interference of moving forks with drug-stabilized topo I-DNA complexes is critical for cell death, in the case of topo II only a partial contribution to cell killing was proposed. We have studied the influence of inhibition of DNA replication by aphidicolin on the production of chromosomal aberrations and SCE by topoisomerase inhibitors. Our results seem to indicate that fork progression is necessary for both cytogenetic endpoints. Pulsed field gel electrophoresis also confirmed this conclusion at the level of DNA breakage (double-strand breaks) efficiently induced by m-AMSA treatment alone, but not when aphidicolin was present. Differences found between topo I and topo II inhibitors (camptothecin and m-AMSA, respectively) could be explained as due to differences in their persistence in trapping the 'cleavable complex'.

Amsacrine↗
PubMed1993

Three-way differentiation of Chinese hamster ovary chromosomes by immunoperoxidase technique using a monoclonal anti-bromodeoxyuridine antibody.

A new permanent staining procedure for sister chromatid differentiation (SCD) in cultured Chinese hamster ovary cells has been established by combining the three-way differentiation in third mitosis (M3) chromosomes and the immunoperoxidase reaction developed with 3,3'-diaminobenzidine using a monoclonal anti-bromodeoxyuridine (BrdU) antibody. This procedure allows SCD at very low BrdU concentrations, and the evaluation of the sister chromatid exchange frequencies on a per cell-cycle basis.

3,3'-Diaminobenzidine↗
PubMed1992

Influence of low doses of BrdU and estimation of spontaneous SCE in CHO chromosomes: three-way differential staining and an immunoperoxidase method.

The influence of low doses of 5-bromodeoxyuridine (BrdU) on the occurrence of sister chromatid exchanges (SCEs) during the first cell cycle, when unsubstituted DNA templates replicate in the presence of the halogenated nucleoside (SCE1) has been assessed in third mitosis (M3) Chinese hamster ovary (CHO) cells showing three-way differential (TWD) staining. In addition, lower concentrations of BrdU, not detectable by Giemsa staining, have been tested by a high resolution immunoperoxidase method (anti-BrdU monoclonal antibody) and SCEs were scored in second mitosis (M2) cells. Our findings was a dose-response curve for SCE1 that allows an estimated mean spontaneous yield of 1.32/cell per cell cycle by extrapolation to zero concentration of BrdU. On the other hand, when the total SCE frequency corresponding to the first and second rounds of replication (SCE1 + SCE2) found in M3 chromosomes was compared with the yield of SCEs scored in M2 cells grown in BrdU at doses lower than 1 microM no further reduction was achieved. This seems to indicate that SCEs can occur spontaneously in this cell line, though the estimated frequency is higher than that reported in vivo.

Animals↗
PubMed1992

Different effectiveness of 4-nitroquinoline-1-oxide, mitomycin C and ethyl methanesulfonate to induce lesions in DNA leading to sister chromatid exchange throughout successive cell cycles in Chinese hamster ovary cells.

The present study was carried out in Chinese hamster ovary cells in order to determine whether lesions induced by three different mutagens, namely 4-nitroquinoline-1-oxide (4-NQO), Mitomycin C (MMC) and Ethyl methanesulfonate (EMS), can persist for more than one cell generation leading to sister chromatid exchanges (SCE) or, alternatively, they are efficiently repaired during the next replicative period after treatment. In order to accurately score the number of SCEs arising during the first (S1), second (S2) and third (S3) DNA synthetic periods, third-cycle (M3) metaphases showing three-way differential (TWD) staining were analyzed. Our results show that, even though the three compounds tested were efficient in increasing the yield of SCE, the frequency of SCE was more dramatically increased after MMC treatment. Differences were also observed among the three mutagens with regard to the persistence of the lesions leading to SCE throughout successive cell generations. EMS-induced lesions appeared as more persistent than those induced by MMC. However, most of the damage induced by the UV mimetic agent 4-NQO seems to be efficiently repaired after the first round of DNA replication following treatment with the drug.

4-Nitroquinoline-1-oxide↗
PubMed1990

Evidence for an adaptive response to radiation damage in plant cells conditioned with X-rays or incorporated tritium.

Allium cepa root-tip cells were first exposed to low 'conditioning' doses of ionizing radiation: to X-rays (0.06 or 0.26 Gy) or to incorporated tritium (1.8 x 10(4) or 7.2 x 10(4) Bq/ml; specific activity: 740.0 GBq/mmol) and subsequently given a 'challenge' dose of 1.5 Gy of X-rays. A reduction in X-ray-induced chromosomal damage was brought about by prior exposure to 0.26 Gy of X-rays, while cells receiving the lower conditioning dose (0.06 Gy of X-rays) did not show any significant reduction. In cells grown in the presence of [3H]TdR on the other hand, the adaptive response was evident after both doses given. The results are essentially in agreement with those published by Wolff's group for human lymphocytes in showing that plant cells in vivo can become 'adapted' by exposure to low-level irradiation so that they become more resistant to the clastogenic effects of X-rays delivered subsequently.

Adaptation, Biological↗
PubMed1990

Adaptive response in human lymphocytes conditioned with hydrogen peroxide before irradiation with X-rays.

Cultured human lymphocytes were first exposed to a low 'conditioning' dose of hydrogen peroxide and, subsequently, irradiated with a 'challenge' dose of 1.5 Gy of X-rays in order to analyse the induction of an adaptive response to oxidative damage. A significant reduction in X-ray-induced chromosome damage was evident when H2O2 was given as a single 30 min pulse 24 h after setting up the cultures and the lymphocytes were exposed to X-rays at 48 h. In contrast, when the cells underwent a repeated exposure to H2O2 before irradiation the yield of aberrations was that expected from the combined treatment.

Cells, Cultured↗
PubMed1989

Scoring of SCE frequency per cell cycle in CHO chromosomes by means of a standardized 3-way-differential staining method.

We have developed 2 alternative protocols to obtain 3-way differentiation (TWD) in Chinese hamster ovary (CHO) cells, in order to analyze the sister-chromatid exchange (SCE) frequency on a per-generation basis. In protocol A, 5-bromodeoxyuridine (BrdU) substitution into DNA was low during the first S period (S1) and high during the next 2 (S2 and S3). In protocol B, on the other hand, BrdU substitution was high during S1 and low during S2 and S3. The main advantage of our procedures is the high efficiency and reproducibility reached by controlling the relative incorporation of BrdU and deoxythymidine (dT) into replicating DNA in the presence of 5-fluorodeoxyuridine (FdU) throughout 3 consecutive S periods. The comparison of the results obtained for both protocols allowed us to evaluate the role of BrdU in the induction of SCEs in each cell cycle. Our results from M3 chromosomes seem to indicate that, under our experimental conditions, neither the concentration of BrdU in the medium nor the rate of BrdU incorporation into nascent DNA is the basic feature responsible for SCE formation. Indeed, the experiments reported here seem to demonstrate that the nature of parental DNA is crucial for the formation of the BrdU-induced SCEs.

Animals↗
PubMed1989

A standardized method for the three-way differential staining of plant chromosomes and the scoring of SCEs per cell cycle.

We have made use of 2 alternative methodologies to obtain 3-way differential staining (TWD) in third-mitosis (M3) chromosomes of Allium cepa, which involve different uptakes of bromodeoxyuridine (BrdU) through 3 cell divisions, in order to evaluate the sister-chromatid exchanges (SCE) frequency on a per-cell-cycle basis. The main innovation introduced by us to the original protocols previously reported has been the use of 5-fluorodeoxyuridine (FdU) to inhibit endogenous thymidine (dT) synthesis. By using different [BrdU]:[dT] ratios in the presence of FdU the relative incorporation of BrdU into replicating DNA can be controlled. The use of 2 different approaches to obtain 3-way differentiation of sister chromatids allowed us to evaluate the role of BrdU in the induction of SCEs in our system. Both procedures rendered nearly 100% of M3 chromosomes showing TWD. An additional advantage of our methodologies is their high degree of reproducibility.

Allium↗
PubMed1989

G2 effects of DNA-repair inhibitors on chromatid-type aberrations in root-tip cells treated with maleic hydrazide and mitomycin C.

In recent years the existence of a DNA-repair process in G2 has been proposed to explain the potentiating effects of DNA-repair inhibitors given in G2 on chromatid aberrations (CA) induced by S-dependent as well as S-independent DNA-damaging agents. In the present report, root-tip cells of Allium cepa were exposed to maleic hydrazide (MH) or mitomycin C (MMC) and post-treated in G2 with caffeine (Caff) and various inhibitors of DNA synthesis. No enhancement of chromosome damage was observed when Caff was present in G2, but hydroxyurea (HU) or 5-fluorodeoxyuridine (FdUrd) potentiated the frequencies of CA. A slight additional increase of CA frequencies was observed following treatment with Ara C and excess thymidine in G2. When MH-damaged cells were pulse-treated with Caff earlier during recovery, the yield of CA was enhanced. The earlier Caff was present following MH treatment, the stronger was the potentiation.

Caffeine↗
PubMed1988

Different distribution of ELI-perikarya in rat mesencephalon using various injection sites for colchicine.

In the present work, using the indirect peroxidase-labelled antibody technique, we made a comparative study about the distribution of Met-enkephalin-like-immunoreactive (ELI)-cell bodies in rat mesencephalon; comparing animals that received colchicine into the lateral ventricle and animals that got the colchicine injected into different cerebral nuclei. We discuss the possibility that the local intracerebral injections are more effective than the intraventricular ones and we show that the results depend on the injection place. So, we think that there are possible met-enkephalinergic connections between the injected region and their corresponding immunoreactive mesencephalic nuclei.

Animals↗
PubMed1987

Effects of caffeine and inhibitors of DNA synthesis on chromatid-type aberrations induced by acetaldehyde in root-tip cells.

Root-tip cells of Allium cepa were exposed to acetaldehyde (AA) and post-treated with caffeine and 3 inhibitors of DNA synthesis, namely hydroxyurea (HU), 5-fluorodeoxyuridine (FdUrd), and arabinofuranosylcytosine (araC). Caffeine strongly potentiated the frequency of chromatid-type aberrations when given immediately after the AA treatment or as a 5-h treatment starting 10 h before the addition of colchicine. In contrast, no enhancement was observed when caffeine was present for the last 2.5 h, simultaneously with colchicine. The inhibitors of DNA synthesis were given following this last schedule. Both HU and FdUrd clearly enhanced the yield of AA-induced chromatid aberrations, while no enhancement of chromosome damage was observed after exposure to araC.

Acetaldehyde↗
PubMed1983

[Ultrastructural study of papillomas of the choroid plexus].

The choroid plexus papillomas are rare neoplasms. They constitute about 0.5 per 100 of all intracranial tumors. From a histological point of view they are generally benign tumors. However they give a high mortality rate due to their location near vital structures, the rapid production of intracranial hypertension, and the rich vascularity making risky the surgical treatment.

Cerebral Ventricle Neoplasms↗