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R L Eberle

Publications and source records attributed to R L Eberle.

9 recordsLinked to original sources
PubMed2004

Complex chromatid-isochromatid exchanges following irradiation with heavy ions?

We describe a peculiar and relatively rare type of chromosomal rearrangement induced in human peripheral lymphocytes that were ostensibly irradiated in G(0) phase of the cell cycle by accelerated heavy ions, and which, to the best of our knowledge, have not been previously described. The novel rearrangements which were detected using mFISH following exposure to 500 MeV/nucleon and 5 GeV/n 56Fe particles, but were not induced by either 137Cs gamma rays or 238Pu alpha particles, can alternatively be described as either complex chromatid-isochromatid or complex chromatid-chromosome exchanges. Different mechanisms potentially responsible for their formation are discussed.

Alpha Particles↗
PubMed2001

Termini of human chromosomes display elevated rates of mitotic recombination.

The strand-specific in situ hybridization technique of CO-FISH was used to probe telomeres of human mitotic cells in order to determine the spontaneous frequency of crossover. This approach allowed the detection of recombinational crossovers occurring anywhere along the length of individual chromosomes, including reciprocal events taking place between sister chromatids. Although the process of sister chromatid exchange (SCE) is the most prominent type of recombination in somatic mammalian cells, our results show that SCEs accounted for less than a third of the recombinational events revealed by CO-FISH. It is concluded that chromosomal regions near the termini of chromosome arms undergo extraordinarily high rates of spontaneous recombination, producing terminal crossovers whose small size precludes detection by standard cytogenetic methods. That similar results were observed for transformed epithelial cells, as well as primary fibroblasts, suggests that the phenomenon is a common characteristic of human cells. These findings are noteworthy because, although telomeric and subtelomeric DNA is known to be preferentially involved in certain types of recombination, the tips of somatic mammalian chromosomes have not previously been identified as preferred sites for crossover. Implications of these results are discussed in terms of limitations imposed on CO-FISH for its proposed use in directional hybridization mapping.

Chromosomes, Human↗
PubMed1998

The effect of maternal position on fetal heart rate during epidural or intrathecal labor analgesia.

OBJECTIVE: This study was designed to determine the relationship between maternal position and the incidence of prolonged decelerations after epidural bupivacaine or intrathecal sufentanil analgesia for labor. STUDY DESIGN: Laboring, healthy, term parturient women, with reassuring fetal heart rate tracings, requesting either epidural (n = 145) or intrathecal (n = 160) analgesia were randomly assigned to lie either supine with measured 30-degree left uterine displacement (n = 136) or in the left lateral decubitus position (n = 145). Patients received either intrathecal sufentanil, 10 microg, or epidural 0.25% bupivacaine, 13 mL. An obstetrician, unaware of patient position or type of anesthesia, examined the fetal heart rate tracings. RESULTS: No demographic differences were noted among the groups. Prolonged decelerations occurred with equal frequency after epidural bupivacaine and intrathecal sufentanil (3.9%). Prolonged decelerations were not related to maternal position. No emergency cesarean deliveries were performed as a result of prolonged decelerations. Prolonged decelerations correlated with the frequency of contractions before induction of analgesia (P < .05). Fewer fetal heart rate accelerations were noted after intrathecal sufentanil than after epidural bupivacaine (P < .005). More ephedrine was used after epidural bupivacaine (P < .001). Patients who received epidural analgesia in the left lateral position were more likely to have an asymmetric block (P < .05). CONCLUSIONS: The risk of prolonged deceleration after epidural bupivacaine or intrathecal sufentanil labor analgesia is unrelated to maternal position or analgesic technique.

Analgesia, Epidural↗
PubMed1996

Labour analgesia. A risk-benefit analysis.

The pain associated with labour can be severe. The ideal labour analgesic does not exist and systemic opioids provide little relief. Nausea, vomiting and sedation are common adverse effects of systemic opioids. Paracervical block can relieve only the pain of the first stage of labour. The duration of analgesia obtained using paracervical block is limited and repeat blocks increase the risk of direct fetal injection. Epidural analgesia effectively relieves labour pain. The insertion of an epidural catheter can provide continuous analgesia throughout labour. In addition, the catheter can be used to provide surgical anaesthesia, should operative delivery be required. Epidural local anaesthetics commonly produce maternal hypotension and motor blockade. However, opioids potentiate the effect of epidural local anaesthetics. Thus, concomitant epidural opioid injection allows the use of lower concentrations of local anaesthetics, decreasing the frequency and severity of hypotension and motor blockade. Epidural analgesia has other, potentially catastrophic, adverse effects but, with safe clinical practice, these problems are extremely rare. Intrathecal injection of opioids or local anaesthetics also effective labour analgesia. However, no single intrathecal drug or drug combination reliably provides analgesia for the duration of labour. Many clinicians use both intrathecal and epidural analgesia as a combined spinal-epidural technique. This approach provides the rapid onset of intrathecal drugs and the flexibility of continuous epidural block. Fetal heart rate decelerations occasionally follow the use of any of the above labour analgesic techniques. Most studies of the aetiology of fetal heart rate decelerations have focused on factors unique to each analgesic technique. However, the similar timing and appearance of fetal bradycardia suggests a common cause. Induction of maternal analgesia may transiently alter the balance between factors encouraging and inhibiting uterine contraction. A temporary increase in the uterotonic effects of endogenous or exogenous oxytocin may then produce a tetanic uterine contraction with subsequent decrease fetal oxygen delivery and resultant fetal bradycardia. Regardless of aetiology, these bradycardias are transient and should not produce maternal or fetal morbidity. Much controversy surrounds the effects of analgesia, especially epidural block, on the course and outcome of labour. Various studies have reported that epidural analgesia slows labour, increases the incidence of malposition of the fetal head, increases the need for forceps delivery and increases the risk of caesarean delivery. Most of the studies reporting these effects are retrospective and nonrandomised. More careful studies suggest that specific anaesthetic techniques (i.e. local anaesthetic-opioid mixtures) or obstetrical management can limit or eliminate these 'risks' of epidural labour analgesia.

Adult↗
PubMed1994

Sequestration of doxorubicin in vesicles in a multidrug-resistant cell line (LZ-100).

A multidrug-resistant Chinese hamster cell line, LZ-8, was subcultured in increasing levels of doxorubicin (DOX) until capable of growth in 100 micrograms/mL DOX. This new derivative, designated LZ-100, is the most DOX-resistant line in the LZ series, based on a comparison of Ki-1 values from cell survival studies. This increased level of drug resistance in LZ-100 cells did not result from (i) higher levels of P-glycoprotein (P-gp) in the plasma membrane compared with LZ-8 cells, since this protein constitutes approximately 20% of the total plasma membrane protein in both cell lines, or (ii) more efficient drug pumping by the same amount of P-gp, since efflux of rhodamine 123 and DOX was comparable in the two cell lines. However, an altered drug distribution was observed in LZ-100 cells compared with wild-type V79 cells; in LZ-100 cells DOX was largely excluded from the nucleus and was sequestered in vesicles in the cytoplasm. The number of vesicles per cell seen after DOX exposure corresponded with the level of drug resistance achieved by the LZ cell lines studied. DOX concentration-response experiments revealed that vesicle formation exhibited a biphasic relationship, with an initial rapid increase followed by a plateau where no further increase was observed. Time-course studies in LZ-100 cells revealed that the maximum number of DOX-containing vesicles per cell occurred 3-4 hr following initiation of DOX treatment. Radiation exposure (10 Gy) immediately preceding DOX treatment decreased the number of vesicles formed in LZ-100 cells by more than one-half and altered the subcellular distribution of DOX from an almost exclusively cytoplasmic to a homogeneous nuclear/cytoplasmic distribution. This redistribution was not a result of radiation inhibition of P-gp efflux. The inhibitory effect of radiation on vesicle formation increased with increasing radiation dose up to 10 Gy. Drug-containing vesicles were also observed in LZ-100 cells following exposure to mitoxantrone or daunorubicin (to which LZ-100 cells are also resistant), but fewer vesicles were observed than with DOX. These studies demonstrate that the drug sequestration phenomenon (i) occurs in cells exhibiting widely different levels of drug resistance, (ii) correlates with the level of drug resistance in LZ cell lines, (iii) occurs rapidly following exposure to DOX, mitoxantrone, or daunorubicin, and (iv) can be inhibited by irradiation.(ABSTRACT TRUNCATED AT 400 WORDS)

ATP Binding Cassette Transporter, Subfamily B, Mem↗
PubMed1992

Characterization of adriamycin-resistant and radiation-sensitive Chinese hamster cell lines.

A series of cell lines derived from Chinese hamster V79 cells by selection in increasing concentrations of Adriamycin (ADRM) was developed to study the mechanisms of drug resistance and its relationship to radiation response. Survival studies revealed that selection in increasingly higher concentrations of ADRM positively correlated with increased cellular drug resistance. Increased cellular resistance correlated positively with amplification of the hamster multidrug-resistance gene (pgp 1) as detected with dot blot analysis using the pCHP1 probe. Southern blot analysis of restriction endonuclease digested DNA (Eco RI, Hind III, Pst I, or Bam HI) showed that (1) some fragments were preferentially amplified compared to others in the ADRM-resistant lines; and (2) no major gene rearrangement appeared to have occurred during the selection for greater ADRM resistance. Levels of pgp 1 gene expression assayed with dot blot and Northern analysis showed a parallel increase of mRNA with gene amplification and increased ADRM resistance. The amounts of the pgp 1 gene product, P-glycoprotein (P-gp), in the cell membrane of the ADRM-resistant cells correlated with the amount of gene amplification/expression. However, levels of P-gp only correlated with degree of drug resistance as measured by cell survival in earlier selection stages (77A and LZ-3). In later selection stages (LZ-8 and LZ-24), higher levels of ADRM resistance were achieved but levels of P-gp did not increase beyond approximately 20% of plasma membrane proteins. These results suggest that (1) the LZ cell plasma membrane may have a physical limit as to the amount of P-gp it can accommodate and/or there is a cellular mechanism for regulating the amount of P-gp in the plasma membrane, and (2) additional resistance mechanisms are present in LZ-8 and LZ-24 cells. Microscopic observations of intracellular drug distribution in these cell lines revealed that (1) ADRM appeared to be sequestered in cytoplasmic vesicles, and (2) the amount of sequestration (number of vesicles) exhibited correlated with the degree of drug resistance attained by the cell lines. These results suggest that drug sequestration is another mechanism of resistance in LZ cells in addition to P-gp-mediated drug efflux.

ATP Binding Cassette Transporter, Subfamily B, Mem↗
PubMed1991

Dot-blot hybridization: quantitative analysis with direct beta counting.

The suitability of using direct beta counting (DBC) for quantitating radioactivity of the probe:target complex in dot-blot hybridization was evaluated using a Packard Matrix 96. A comparison of blots analyzed using autoradiography followed by densitometry scanning (film/densitometry) with those analyzed using direct beta counting revealed similar data trends with the two methods. However, direct beta counting quantitated the amount of radioactivity in the dot blots directly (without film exposure or additional sample preparation), which significantly reduced the time required to obtain results. Blots analyzed first with direct beta counting and then liquid scintillation counting exhibited similar data trends with both methods. Despite a decreased counting efficiency, analysis with direct beta counting has the following advantages compared with liquid scintillation counting: 1) no additional sample preparation is required (no vials or cocktail are used), 2) no sample destruction occurs due to analysis and 3) quantitative results are obtained more rapidly (since the radioactivity for all 96 samples in a dot blot is simultaneously determined in real time). Analysis with direct beta counting was also shown not to interfere with the successful reprobing of stripped dot blots with either unique sequence or total genomic probes. Overall, direct beta counting provides quick, quantitative results for dot blots while saving considerable time and effort.

Animals↗
PubMed1991

Interaction between radiation and drug damage in mammalian cells. V. DNA damage and repair induced in LZ cells by adriamycin and/or radiation.

A multi-drug-resistant cell line selected in increasing concentrations of Adriamycin and designated LZ (J. A. Belli, Radiat. Res. 119, 88-100, 1989) is shown to exhibit a survival response characterized by radiation sensitivity and Adriamycin resistance. To determine if this response is due to alterations in either the initial levels of damage induced or the repair of DNA damage, LZ cells and the parental V79 cells were exposed to either radiation or Adriamycin and the damage and repair were measured with alkaline or nondenaturing filter elution. After exposure to radiation, induction and repair of both single-strand and double-strand breaks were equivalent. LZ cells exposed to 100 micrograms/ml Adriamycin for 1 h contained no measurable damage while the same treatment induced breaks and crosslinks in V79 cells. Pretreatment of LZ cells for 1 h with Adriamycin before irradiation did not alter either the initial levels of induced damage or the repair of strand breakage. These results suggest that (1) mechanisms other than differential induction and repair of strand breaks are responsible for the increased radiation sensitivity in LZ, and (2) the lack of Adriamycin-induced DNA damage in LZ is at least partially responsible for the increased cell survival after treatment.

Animals↗