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Biomedical subjects

S Grant

Publications and source records attributed to S Grant.

At least 199 records · Page 11Linked to original sources

The effects of halothane and nitrous oxide on the pharmacokinetics of propofol in dogs.

The pharmacokinetics of propofol, 6.5 mg/kg, administered as a bolus dose intravenously (i.v.) were studied in six dogs (group 1). The effect of maintaining anaesthesia with halothane and nitrous oxide in oxygen on propofol pharmacokinetics was also investigated in six dogs undergoing routine anaesthesia (group 2). Induction of anaesthesia was rapid in all animals. Post-induction apnoea was a feature in three of the 12 dogs. The blood propofol concentration-time profile was best described by a bi-exponential decline in two dogs in group 1 and in 3 dogs in group 2, and by a tri-exponential decline in four dogs in group 1 and 3 dogs in group 2. The elimination half-life was long in both groups (90.9 min and 75.2 min, respectively), the volume of distribution at steady state large (4889 and 4863 ml/kg) and the clearance rapid (58.6 and 56.3 ml/kg.min). There were no significant differences between the groups, thus indicating that maintenance of anaesthesia with halothane and nitrous oxide had no effect on the pharmacokinetics of propofol in the dog.

Anesthesia↗

Physiological and psychological responses to a university fitness session.

The purpose of this study was to examine the physiological and psychological responses to a university fitness session entitled 'popmobility'. A popmobility session consists of 20 min of aerobic activities, 5 min of local muscular endurance exercises and 5 min of flexibility exercises. Ten regular participants of these sessions, women of mean(s.d.) age 21.2(1.5) years, took part in the study. A maximal oxygen uptake (VO2max) treadmill test was performed by each subject to obtain VO2max and maximum heart rate values. In a laboratory, heart rate and VO2 were measured throughout a popmobility session for each subject. Rate of perceived exertion (RPE) was measured every 5 min throughout the session. The mean intensity of the aerobic part of the session ranged from 67.7-82.6% of the subject's VO2max (mean of 76.4% VO2max). The mean heart rate reserve for the aerobic section was 75.6%. While the relative oxygen consumption remained fairly static during the aerobic section, the RPE score rose. The mean(s.d.) total energy expenditure was 236.6(28.4) kcal (range 203-288). The popmobility session is of adequate intensity to improve the aerobic fitness of its participants. Heart rate, as used as a measure of intensity during a popmobility session, would appear to be a fairly accurate indicator of intensity. However, the use of RPE for exercise prescription in popmobility sessions is inappropriate. Popmobility could also be useful in a weight-reduction programme.

Adult↗

Modulation of the activity of a human granulocyte-macrophage colony-stimulating factor/interleukin-3 fusion protein (pIXY 321) by the macrocyclic lactone protein kinase C activator bryostatin 1.

We have examined the effect of the macrocyclic lactone protein kinase C (PK-C) activator bryostatin 1 on the proliferative capacity and lineage commitment of CD34+ human bone marrow cells exposed to the granulocyte-macrophage colony-stimulating factor/interleukin-3 (GM-CSF/IL-3) fusion protein pIXY 321. pIXY 321 administered at a dose of 10 ng/mL was as effective as the combination of plateau concentrations of recombinant (r) IL-3 and rGM-CSF (e.g., 50 ng/mL) in stimulating the growth of day-14 committed myeloid progenitors (colony-forming units granulocyte/macrophage [CFU-GM]). In the large majority of samples tested, coadministration of 0.5 to 100 nM bryostatin 1 with either pIXY 321 or the combination of rIL-3 plus rGM-CSF led to modest but significant increases (e.g., 30 to 75%) in the number of CFU-GM, compared to administration of growth factors alone. The degree of bryostatin 1-induced potentiation, however, was considerably less than that previously observed in the case of cells exposed to either rIL-3 or rGM-CSF, where increases of 100 to 150% were regularly noted. While at least 50% of day-14 CFU-GM stimulated by either pIXY 321 or the combination of rIL-3 plus rGM-CSF were of the pure or mixed eosinophilic variety, coadministration of bryostatin 1 resulted in a dramatic inhibition of eosinophilic colonies and a corresponding increase in pure and mixed neutrophil and macrophage colonies. Although coadministration of recombinant granulocyte colony-stimulating factor (rG-CSF) or recombinant colony-stimulating factor-1 (rCSF-1) mimicked the capacity of bryostatin 1 to increase the total number of pIXY 321-induced day-14 CFU-GM, these growth factors, unlike bryostatin 1, were not capable of inhibiting eosinophilic colony formation. Furthermore, whereas addition of neutralizing antibodies to G-CSF or CSF-1 blocked the capacity of these growth factors to potentiate colony formation in the presence of pIXY 321, it did not abrogate the effect of bryostatin 1 on progenitor cell growth or lineage commitment. Finally, in contrast to its effects on committed myeloid progenitors, bryostatin 1 did not increase the growth of erythroid (burst-forming units-erythroid [BFU-E]) and multipotent (multipotent colony-forming units [CFU-GEMM]) progenitors stimulated by pIXY 321, but instead inhibited colony formation at higher concentrations (e.g., 10 to 100 nM).(ABSTRACT TRUNCATED AT 400 WORDS)

Antibodies↗

Phase I study of a continuous infusion of high-dose ara-C in conjunction with a fixed dose of 2'-deoxycytidine (IND 28108) in patients with refractory leukemia: an interim report.

A total of 16 patients with refractory leukemia have been entered on a phase I study employing escalating doses of a 96 h continuous infusion of high-dose cytosine arabinoside (ara-C) administered in conjunction with a 120 h infusion of a fixed dose (22 g/m2 per day) of 2'-deoxycytidine (IND 28108) as a host-protective agent. Extramedullary toxicities, even at the highest ara-C dose level (e.g. 14 g/m2 per day) were mild (e.g. CALGB grade I or II), and consisted of diarrhea, fluid retention, somnolence, hepatic dysfunction, and febrile episodes. Neutropenia and thrombocytopenia were noted in all patients, but no serious infections or bleeding episodes were encountered. Steady state plasma ara-C concentrations of 20-60 microM were observed in patients treated at the 14 g/m2 dose, and were associated with plasma ara-U concentrations approaching 1 mM. Although no complete remissions were obtained at the ara-C dose levels tested to date, three partial remissions were noted, and the large majority of patients experienced a clearing of peripheral blood blasts. In addition, several patients who failed to achieve objective responses exhibited atypically benign clinical courses following completion of therapy. These findings demonstrate that 2'-deoxycytidine protects humans from otherwise lethal doses of high-dose ara-C administered by continuous infusion, and substantially ameliorates the hematologic and non-hematologic toxicity of such regimens without completely abrogating antileukemic activity. Determination of the ultimate efficacy of this strategy will require identification of an ara-C maximum tolerated dose and evaluation of the antileukemic potential of this regimen in prospective phase 11 trials.

Adult↗

Granulocyte-macrophage colony-stimulating factor/interleukin-3 fusion protein (pIXY 321) enhances high-dose Ara-C-induced programmed cell death or apoptosis in human myeloid leukemia cells.

High dose Ara-C (HIDAC) induces programmed cell death (PCD) or apoptosis in vitro in human myeloid leukemia cells, which correlates with the inhibition of their clonogenic survival. Hematopoietic growth factors (HGFs) granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-3 (IL-3) have been demonstrated to enhance the metabolism and cytotoxic effects of HIDAC against leukemic progenitor cells. We examined the effect of pIXY 321 (a GM-CSF/IL-3 fusion protein) on HIDAC-induced PCD and related gene expressions as well as HIDAC-mediated colony growth inhibition of human myeloid leukemia cells. Unlike the previously described effects of HGFs on normal bone marrow progenitor cells, exposure to pIXY 321 alone for up to 24 hours did not suppress PCD in HL-60 or KG-1 cells. However, exposure to pIXY 321 for 20 hours followed by a combined treatment with Ara-C plus pIXY 321 for 4 or 24 hours versus treatment with Ara-C alone significantly enhanced the oligonucleosomal DNA fragmentation characteristic of PCD. This was temporally associated with a marked induction of c-jun expression and a significant decrease in BCL-2. In addition, the treatment with pIXY 321 plus HIDAC versus HIDAC alone produced a significantly greater inhibition of HL-60 colony growth. These findings highlight an additional mechanism of HIDAC-induced leukemic cell death that is augmented by cotreatment with pIXY 321 and may contribute toward an improved antileukemic activity of HIDAC.

Actins↗

Bryostatin 1 modulates the proliferation and lineage commitment of human myeloid progenitor cells exposed to recombinant interleukin-3 and recombinant granulocyte-macrophage colony-stimulating factor.

The activity of protein kinase C (PK-C) has been implicated in the regulation of the growth and differentiation of both normal and neoplastic hematopoietic cells. We have examined the effects of the PK-C-activating agents phorbol 12,13-dibutyrate (PDBu), mezerein, and bryostatin 1 on the proliferation and lineage commitment of CD34+ human myeloid progenitor cells stimulated by recombinant interleukin-3 (rIL-3) and/or recombinant granulocyte-macrophage colony-stimulating factor (rGM-CSF). Although each of the PK-C activators administered alone induced no colony formation, coadministration of these agents with plateau concentrations of each cytokine (eg, 50 ng/mL) increased the number of day 14 granulocyte-macrophage colony-forming units by 100% to 150%. The number of pure and mixed neutrophil and macrophage colonies was substantially enhanced in the presence of PK-C activators, whereas the percentage and, in most cases, the absolute number of eosinophilic colonies was significantly reduced. The inhibition of eosinophilic colony formation was not overcome by the addition of rIL-5. Although addition of bryostatin 1 24 hours before rIL-3 abrogated the increase in total colony formation observed with simultaneous administration of factors, the inhibition of eosinophilic colonies and the increase in neutrophil/macrophage colonies persisted under these conditions. The addition of bryostatin 1 for up to 144 hours after rIL-3 continued to potentiate total colony formation, whereas the inhibition of eosinophilic commitment was lost after 120 hours. Together, these results suggest that pharmacologic interventions at the level of PK-C may regulate both the proliferation as well as the lineage commitment of human hematopoietic progenitors exposed to rGM-CSF and rIL-3.

Bryostatins↗

Potentiation of the activity of 1-beta-D-arabinofuranosylcytosine by the protein kinase C activator bryostatin 1 in HL-60 cells: association with enhanced fragmentation of mature DNA.

We have examined the interaction between 1-beta-D-arabinofuranosylcytosine (ara-C) and the macrocyclic lactone protein kinase C activator bryostatin 1 in the human promyelocytic leukemia cell line HL-60. Preexposure of cells to 10 nM bryostatin 1 for 24 h, followed by an additional 24-h incubation with 10 microM ara-C, resulted in greater than additive inhibitory effects toward clonogenic HL-60 cells. In a series of alkaline elution assays, cells preincubated with bryostatin 1 and prelabeled with [3H]thymidine exhibited a significant increase in DNA fragmentation following exposure to ara-C in comparison to cells exposed to ara-C alone. This increase in DNA damage was apparent at both neutral and alkaline pH and was not protein associated. In contrast, studies using cells pulse-labeled with [3H]thymidine immediately before analysis suggested that bryostatin 1 pretreatment did not increase the ability of ara-C to interfere with DNA replicative intermediates. Additional studies demonstrated that the increase in DNA fragmentation induced by bryostatin 1 and ara-C preceded both loss of cell membrane integrity (as determined by trypan blue exclusion) as well as depletion of intracellular ATP and NAD pools. Furthermore, the enhanced inhibitory effects of bryostatin 1 and ara-C toward clonogenic HL-60 cells did not appear to result from the induction of cellular differentiation. Finally, agarose gel electrophoresis of DNA obtained from cells exposed to both bryostatin 1 and ara-C revealed a pattern of integer multiples of 180- to 200-base pair fragments commonly associated with endonucleolytic cleavage; the extent of this fragmentation was considerably greater than that observed in cells exposed to ara-C alone. Taken together, these findings suggest that exposure of HL-60 cells to bryostatin 1 renders them more susceptible to ara-C-related DNA damage and that this phenomenon contributes to the cytotoxic effects of this drug combination. They also raise the possibility that bryostatin 1, perhaps through modulation of intracellular signaling events in leukemic cells, has the capacity to potentiate ara-C-related apoptosis or programmed cell death.

Adenosine Triphosphate↗

A topographic representation of auditory space in the external nucleus of the inferior colliculus of the guinea-pig.

The possibility that the external nucleus of the inferior colliculus (ICX) of the pigmented guinea-pig contains a map of auditory space has been investigated. Auditory stimuli consisted of broad-band sound delivered under free-field anechoic conditions from a range of positions around the animal's azimuthal axis. The responses of clusters of neurons in the ICX to threshold and to near-threshold stimuli displayed sharp spatial tuning. The responses recorded from rostral ICX revealed a preference for auditory stimuli in the anterior field while more caudal neurons preferentially responded to sounds presented in the posterior field. Neurons at intermediate points, along the rostro-caudal axis of the nucleus, displayed preferences for sound stimuli in appropriately intermediate field positions along the contralateral azimuthal axis. At higher stimulus intensities the spatial tuning of the responses decreased, but the optimal direction of preference was usually retained. The contribution of binaural processing to auditory spatial tuning was evident, since unilateral cochlea ablation destroyed the spatial tuning at higher stimulus intensities. The results presented provide the first evidence that a topographically ordered representation of the contralateral auditory azimuth is present in the ICX of a mammal.

Acoustic Stimulation↗

Industrial blood lead levels in the South Island during 1988 and 1989: trends and follow up patterns.

AIMS: to assess trends in industrial lead exposure and the monitoring programmes in the South Island of New Zealand. METHODS: during the period 1 January 1988 to 31 December 1989, industrial lead exposure was analysed in 1425 workers in at risk occupations and the efficiency of retesting programmes was determined. RESULTS: forty-four percent of these workers had red cell lead levels above 1.9 mumol/L, the top of the reference range for an unexposed population, and 71 individuals had levels exceeding the recommended action limits (males greater than 5.0 mumol/L, and females greater than 3.8 mumol/L). Although most occupational groups showed a small decline in mean red cell lead levels, the pattern of exposure was similar to previous reports. On average, only 43% of exposed workers were retested within the recommended period and 32% of these workers were not retested within 2 years of having a raised blood lead level. CONCLUSIONS: retesting was inefficient but was most reliable when industrial health nurses were employed for monitoring. Not all lead poisoning comes from the traditional lead based industries and significant decreases were found in workers whose primary exposure is to lead from petrol.

Aftercare↗

The mosaic of alpha cells in the cat retina is not dependent on axon terminal interactions during development.

Alpha ganglion cells in the cat retina are distributed in a regular array. It has been proposed that the development of this mosaic pattern is achieved by class-specific interactions between the dendrites and/or axon terminals of neighbouring alpha cells, but the relative contributions that are made by each of these factors to the regularity of the mosaic remain unclear. An opportunity to address this question is provided by a comparative study of the distribution of alpha cells across the nasotemporal division of normally pigmented and Siamese cat retinae. In this strip of retina the alpha cell population divides its axon terminals between the two sides of the brain. Hence, the potential for interactions between axon terminals of alpha cells whose somata are adjacent within this region of the retina is undermined. This situation is exacerbated in Siamese cats because they have a congenital abnormality in this region, which results in an abnormally wide nasotemporal division. In this study the regularity of the alpha cell mosaic has been analysed at a wide range of eccentricities, including the nasotemporal division. In normally pigmented cats the nasotemporal division was visualised directly following unilateral horseradish peroxidase injections into the thalamus. In Siamese cats, the location of the abnormally wide nasotemporal division was inferred from electrophysiological recordings undertaken in their striate cortices. In both strains of cat the regularity of the alpha cell mosaic was shown to be largely independent of their density or retinal location. Furthermore, in the normals, the separate mosaics for ipsilaterally or contralaterally projecting alpha cells within the nasotemporal division were found to be no more or less regular than that for the combined population. This finding implies that there is no disturbance in their order across the discontinuity formed by their separate chiasmatic routes. Similar results were obtained from within the abnormal nasotemporal division of the Siamese cat. It is concluded that terminal interactions play an insignificant role in the formation of the adult alpha cell mosaic, the regularity of which is probably established by class-specific intraretinal interactions.

Animals↗

Changing patterns of binocular visual connections in the intertectal system during development of the frog, Xenopus laevis. III. Modifications following early eye rotation.

In the frog, Xenopus laevis, a system of intertectal connections underlies the visual projection from an eye to its ipsilateral tectal lobe and is involved in the topographic representation of binocular visual space. Rotation of one eye in early life may be followed by a radical rearrangement of the connections in this system. The modified pattern which later emerges is that which keeps the visual projection through the ipsilateral eye in topographic registration with the direct visual projection from the contralateral eye to the same tectal lobe. This plasticity requires visual experience. In this paper we describe the time-course and sequence of events by which this plasticity is effected. Following rotation of one eye in larval animals or in animals undergoing metamorphic climax, the earliest evidence of intertectal modification was found 3-4 weeks after metamorphosis. With increasing intervals after metamorphosis an increasing proportion of animals displayed modified intertectal systems. At intermediate intervals many animals showed partial modifications, which were interpreted as transitional stages in the modification process. Analysis of these transitional stages indicated that the sequence of events involved in the elaboration of a modified intertectal system following the experimental alteration of eye alignment exhibits features in common with rearrangements of the system that occur during normal development in response to growth-related alterations in eye alignment.

Animals↗

Plasticity of binocular visual connections in the frog, Xenopus laevis: reversibility of effects of early visual deprivation.

In frogs, each lobe of the midbrain optic tectum receives two spatially aligned maps of binocular visual space, one through each eye. The map from the contralateral eye is the product of direct retinal input to the tectum, while that from the ipsilateral eye is mediated by a commissural system of connections that relays binocular visual information from one tectal lobe to the other. In Xenopus laevis, dark-reared from early life, the visuotectal map through the ipsilateral eye exhibits significant signs of disorder indicating that the normal development of the underlying "intertectal" system involves experience-dependent processes. Here we investigate the potential of this system for recovery from such deprivation-induced disruption. Three groups of animals were first dark-reared from embryonic stages until 3 months, 12 months or 2 years after metamorphosis. These short-, medium- and long-term periods of visual deprivation are associated with a progressive disturbance of the intertectal system (Grant and Keating 1989b). The animals then experienced 12-18 months in a normal visual environment, before an electrophysiological mapping experiment was carried out to reveal the status of their intertectal system. Quantitative analyses of the visuotopic order in the ipsilateral visuotectal map and of its spatial alignment with the map derived from the contralateral eye revealed that these features were completely normal in all 3 experimental groups. It is concluded that defects present in the intertectal system of dark-reared Xenopus can be repaired by subsequent visual experience, and that the reparative capacity is not limited either by the animal's age or by the existing degree of prior disruption.

Animals↗

Topographic organization, number, and laminar distribution of callosal cells connecting visual cortical areas 17 and 18 of normally pigmented and Siamese cats.

The callosal connections between visual cortical areas 17 and 18 in adult normally pigmented and "Boston" Siamese cats were studied using degeneration methods, and by transport of WGA-HRP combined with electrophysiological mapping. In normal cats, over 90% of callosal neurons were located in the supragranular layers. The supragranular callosal cell zone spanned the area 17/18 border and extended, on average, some 2-3 mm into both areas to occupy a territory which was roughly co-extensive with the distribution of callosal terminations in these areas. The region of the visual field adjoining the vertical meridian that was represented by neurons in the supragranular callosal cell zone was shown to increase systematically with decreasing visual elevation. Thus, close to the area centralis, receptive-field centers recorded from within this zone extended only up to 5 deg into the contralateral hemifield but at elevations of -10 deg and -40 deg they extended as far as 8 deg and 14 deg, respectively, into this hemifield. This suggests an element of visual non-correspondence in the callosal pathway between these cortical areas, which may be an essential substrate for "coarse" stereopsis at the visual midline. In the Siamese cats, the callosal cell and termination zones in areas 17 and 18 were expanded in width compared to the normal animals, but the major components were less robust. The area 17/18 border was often devoid of callosal axons and, in particular, the number of supragranular layer neurons participating in the pathway were drastically reduced, to only about 25% of those found in the normally pigmented adults. The callosal zones contained representations of the contralateral and ipsilateral hemifields that were roughly mirror-symmetric about the vertical meridian, and both hemifield representations increased with decreasing visual elevation. The extent and severity of the anomalies observed were similar across individual cats, regardless of whether a strabismus was also present. The callosal pathway between these visual cortical areas in the Siamese cat has been considered "silent," since nearly all neurons within its territory are activated only by the contralateral eye. The paucity of supragranular pyramidal neurons involved in the pathway may explain this silence.

Albinism↗

Effects of bryostatin 1 and rGM-CSF on the metabolism of 1-beta-D-arabinofuranosylcytosine in human leukaemic myeloblasts.

The effects of the protein kinase C activator bryostatin 1, either with or without recombinant granulocyte-macrophage colony stimulating factor (rGM-CSF) were examined with respect to the in vitro metabolism of ara-C in leukaemic myeloblasts obtained from 10 patients with acute myelogenous leukaemia (AML). Coincubation of cells with 12.5 x 10(-9) M bryostatin 1 and 10(-5) M ara-C for 4 h resulted in a significant increase in ara-CTP formation (compared to controls) in 6/10 specimens (mean increase 106%; range 38-255%), and no change in the remainder. In contrast, coincubation of cells with 1.25 ng/ml rGM-CSF resulted in a significant increase in only one specimen, and decreases in two. Bryostatin 1 also significantly increased ara-C DNA incorporation in 6/9 evaluable samples, including two which did not display an increase in ara-CTP formation. Coincubation of cells with both bryostatin 1 and rGM-CSF did not lead to a further increase in ara-CTP formation or ara-C DNA incorporation compared to values obtained with either agent alone. Finally, exposure of blasts to bryostatin 1 for 24 h before ara-C led to an increase in ara-CTP formation in 3/8 additional specimens, and a decrease in one sample displaying evidence of bryostatin 1-induced macrophage differentiation. Incubation of cells with both rGM-CSF and bryostatin 1 for this period resulted in ara-CTP levels equivalent to those obtained with bryostatin 1 alone. These studies indicate that while bryostatin 1 exerts a heterogeneous effect on ara-C metabolism in leukaemic myeloblasts, it is capable of potentiating ara-C phosphorylation in a subset of patient samples, including some that do not exhibit an increase in response to rGM-CSF. They also raise the possibility that bryostatin 1-induced potentiation of ara-C metabolism in some leukaemic cells may contribute, at least in part, to the antileukaemic efficacy of this drug combination.

Antineoplastic Agents↗

The effects of a university fitness programme on health-related variables in previously sedentary males.

This study reports on the effects of a 10-week university fitness programme on health-related fitness variables. Twenty-one male exercisers, aged 37.0(10.3) years (mean(s.d.); range 21-58), and 22 male controls, aged 38.6(7.9) years (mean(s.d.); range 17-54), volunteered to take part. Two sample t-tests and 95% confidence intervals were used to determine if the exercise group demonstrated a greater average improvement than the control group and the average improvement in both groups separately. The exercise group showed a greater average improvement over the controls from Test 1 (before fitness programme) to Test 2 (after) in the following: steady-state heart rate (beats min-1) 95% confidence intervals (-7.8, -16.2); predicted VO2max (ml kg-1 min-1): 95% confidence intervals (3.2, 6.6); sit-ups (repetitions): 95% confidence intervals (3.1, 7.0); flexibility (cm): 95% confidence intervals (3.3, 6.9). There was no significant difference between the exercise group and control group in body weight, percentage body fat, blood pressure, total plasma cholesterol, high-density lipoprotein and triglycerides. The exercise programme improved aerobic fitness, local muscular endurance and flexibility. However, the increase in aerobic fitness did not coincide with beneficial changes in the coronary risk profile.

Adolescent↗

Altered expression of cell cycle related genes including c-myb in a subclone of HL-60 that exhibits reversible differentiation.

A differentiation resistant subclone of HL-60, DMSOr, was removed from the selective pressure of dimethyl sulfoxide and characterized with a new stable phenotype of reversible differentiation. DMSOr cells, when treated with 1.25% dimethyl sulfoxide, differentiated in a manner similar to the parental HL-60 with respect to morphological changes, increase in superoxide production, and withdrawal from cell cycle. Upon removal of the dimethyl sulfoxide at points normally associated with commitment to terminal differentiation, DMSOr reverted to the immature phenotype. This demonstrates an uncoupling of the morphological, functional, and antiproliferative effects of differentiation from commitment to terminal differentiation. Associated with the reversible phenotype of DMSOr was an altered expression of the c-myb oncogene. In HL-60, c-myb expression was down-regulated by 72 h and completely diminished by 144 h. Northern blot analysis of DMSOr demonstrated greater levels of expression of c-myb at 72 and 144 h. Similar results were shown with histone H4, cdc2 kinase, and, to a lesser extent, ornithine decarboxylase. The c-myb related gene B-myb did not show altered regulation during differentiation. The results suggest that altered expression of genes that control cell cycle may be critical for the reversible phenotype of DMSOr.

Base Sequence↗

Effect of bryostatin 1 on the in vitro radioprotective capacity of recombinant granulocyte-macrophage colony-stimulating factor (rGM-CSF) toward committed human myeloid progenitor cells (CFU-GM).

We have examined the effect of the macrocyclic lactone protein kinase C (PK-C) activator bryostatin 1 on the in vitro radioprotective capacity of recombinant granulocyte-macrophage colony-stimulating factor (rGM-CSF) toward normal committed myeloid progenitor cells (day-14 granulocyte-macrophage colony-forming units [CFU-GM]). Preincubation of T-cell- and adherent cell-depleted bone marrow mononuclear cells with 12.5 nM bryostatin 1 and either 1.25 or 50 ng/ml rGM-CSF for 24 h resulted in an 18%-30% survival at 4-5 Gy, whereas cells exposed to rGM-CSF alone gave rise to no detectable colonies at radiation doses greater than 2.5 Gy. Coadministration of bryostatin 1 also led to a threefold increase in Do values for both rGM-CSF concentrations. A similar enhancement of radioprotective effects was observed with the tumor-promoting phorbol ester phorbol dibutyrate. Exposure of cells to both bryostatin 1 and rGM-CSF immediately following irradiation also resulted in enhanced progenitor cell survival when compared to rGM-CSF alone, but radioprotective effects were less than those observed when cells were preincubated with these factors. Cells preconditioned with bryostatin 1 and rGM-CSF prior to exposure to 2 or 4 Gy gave rise to significantly more colonies when radiation was administered as a 4-h divided dose, suggesting that bryostatin 1 may act by potentiating rGM-CSF-induced repair of sublethal radiation damage. Finally, pre-exposure of enriched progenitor cells (CD34+) to bryostatin 1 and rGM-CSF resulted in radioprotective effects that were less than those observed for partially purified populations with respect to the total population of surviving myeloid colonies. However, CD34+ cells preincubated with bryostatin 1 and rGM-CSF prior to irradiation exhibited a significant increase in both the percentage and absolute number of neutrophilic and macrophage colonies, and a reduction in eosinophilic colonies, compared to cells exposed to rGM-CSF alone. These studies suggest that bryostatin 1 (and possibly other PK-C activators) potentiates the in vitro radioprotective effects of rGM-CSF and may also regulate the lineage specificity of this response.

Antigens, CD↗