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Biomedical subjects

S Buck

Publications and source records attributed to S Buck.

At least 37 records · Page 2Linked to original sources
PubMed1995

Elevated dihydrofolate reductase and impaired methotrexate transport as elements in methotrexate resistance in childhood acute lymphoblastic leukemia.

A retrospective study of clinical resistance to methotrexate (MTX) was performed on 29 archival specimens of frozen lymphoblasts obtained from children with acute lymphoblastic leukemia (ALL), including 19 at initial presentation and 10 at first relapse. Blasts were assayed for dihydrofolate reductase and MTX transport by flow cytometry using the fluorescent methotrexate analog, PT430 (Rosowsky et al, J Biol Chem 257:14162, 1982). In contrast to tissue culture cells, patient blasts were often heterogeneous for dihydrofolate reductase content. Of the 19 specimens at initial diagnosis, 7 exhibited dual blast populations, characterized by threefold to 10-fold differences in relative dihydrofolate reductase; the dihydrofolate reductase-overproducing populations comprised 12% to 68% of the total blasts for these specimens. Remission duration intervals for patients exhibiting dual blast populations were notably shorter than for patients expressing a single blast population with lower dihydrofolate reductase ( < or = 9 months v > or = 15 months, respectively), a difference that was statistically significant (P = .045). There was no apparent correlation between expression of increased dihydrofolate reductase at diagnosis and known patient and disease prognostic features (immunophenotype, age, sex, and white blood count). For the relapsed patients, 4 of 10 exhibited dual lymphoblast populations with elevated dihydrofolate reductase. The majority of the patient lymphoblast specimens were entirely competent for MTX transport and, likewise, expressed immunoreactive reduced folate carriers by indirect immunofluorescence staining with specific antiserum to the transporter. Three patients (2 at relapse and 1 at diagnosis) exhibited heterogeneous expression of imparied MTX transport (14% to 73% of blasts). In only 1 of these patients did the majority of the lymphoblasts (73%) show impaired MTX transport and for this specimen, immunoreactive carrier proteins were virtually undetectable. These results suggest that heterogeneous expression of elevated dihydrofolate reductase and impaired MTX transport are important modes of resistance in childhood ALL patients undergoing chemotherapy with MTX and that these parameters may serve as predictive indices of clinical response to MTX.

Adolescent↗
PubMed1995

Selection for increased longevity in Drosophila melanogaster: a reply to Lints.

An important tool in the genetic analysis of longevity and aging in Drosophila melanogaster is the use of strains selected directly for late-age reproduction and indirectly for extended longevity. Following some initial failures to select for extended longevity, there are now a number of laboratories which have successfully selected for long life, using the techniques of late-age reproduction as well as selection for stress resistance. Baret and Lints [Gerontology 1993;39:252-259] have recently cast doubt on the reality of a number of these selected strains, including our own, suggesting that the difference in longevity between the long-lived and normal-lived strains disappears when the data are examined as a function of the number of days after the beginning of the selection experiment instead of as a function of the number of generations. With regard to our selected lines, they based their analysis on a subset of the published data dealing with these strains, and which covered 21 generations, or 40 months, of selection. We now present data for over 70 generations, or 155 months, of selection and maintenance. The Baret-Lints hypothesis makes two strong predictions, namely that (1) the longevity difference between the several strains should disappear when the data are replotted according to their fashion, and (2) there should be no other significant biological difference between the strains. Our data falsifies both of these predictions. The Baret-Lints hypothesis is flawed and should be disregarded.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1995

Retirement of senior physicians in rural Minnesota. Factors influencing physicians' plans to retire.

The demand for physicians in rural Minnesota is likely to increase significantly over the next few years. Nearly 20% of all rural physicians in Minnesota are older than age 65. This paper reports the results of a survey of rural Minnesota physicians over age 60 on their current practices and their plans for retirement. Of the 33 physicians questioned, 25 (75%) currently had plans to retire and more than half of the 25 planned to retire in the next two years. Factors that these physicians perceived as important in determining their decisions to retire or continue practice are described, and implications for the delivery of health care in rural communities are discussed.

Aged↗
PubMed1994

Astrocytosis and axonal proliferation in the hippocampus of S100b transgenic mice.

S100 beta is a calcium-binding protein that is expressed at high levels in brain primarily by astrocytes. Addition of the disulfide-bonded dimeric form of S100 beta to primary neuronal and glial cultures and established cell lines induces axonal extension and alterations in astrocyte proliferation and phenotype, but evidence that S100 beta exerts the same effects in vivo has not been presented. An 8.9-kb murine S100b genomic clone was used to produce two lines of transgenic mice in which S100 beta RNA is increased in a dose-related manner to 2-fold and 7-fold above normal. These lines show concomitant increased S100 beta protein throughout the brain. Expression in both lines is cell type- and tissue-appropriate, and expression levels are correlated with the transgene copy number, demonstrating that sequences necessary for normal regulation of the gene are included within the cloned segment. In the hippocampus of adult transgenic mice, Western blotting detects elevated levels of glial fibrillary acidic protein and several markers of axonal sprouting, including neurofilament L, phosphorylated epitopes of neurofilament H and M, and beta-tubulin. Immunocytochemistry demonstrates alterations in astrocyte morphology and axonal sprouting, especially in the dentate gyrus. Thus, both astrocytosis and neurite proliferation occur in transgenic mice expressing elevated levels of S100 beta. These transgenic mice provide a useful model for studies of the role of S100 beta in glial-neuronal interactions in normal development and function of the brain and for analyzing the significance of elevated levels of S100 beta in Down syndrome and Alzheimer disease.

Animals↗
PubMed1994

[Pilot study of the effect of interferon-alpha on atopic eczema].

In this pilot study 12 patients with moderate to severe atopic eczema were treated with 2 million IU interferon alpha 2a (Roferon A) three times weekly for 8 weeks and followed up for a further 10 weeks: 2 patients showed clear and 9 a slight to moderate reduction of their skin lesions; 1 patient got worse. Pruritus did not decrease and even increased in a few patients, and IgE levels showed no change during and after treatment. All patients noted mild, transient, flu-like symptoms. The efficacy of interferon alpha in the present therapeutic design for the treatment of atopic eczema must thus be classified as only moderate.

Adult↗
PubMed1993

Targeting of SIR1 protein establishes transcriptional silencing at HM loci and telomeres in yeast.

Previous studies suggest that the yeast SIR1 protein is involved in the establishment of transcriptional silencing at the HM mating-type loci. Here we show that a GAL4 DNA-binding domain-SIR1 hybrid protein (GBD-SIR1), when targeted to an HMR locus containing GAL4-binding sites (UASG), can establish silencing and bypass the requirement for the silencer element HMR-E. Silencing mediated by GBD-SIR1 requires the trans-acting factors that normally participate in repression, namely, SIR2, SIR3, SIR4, and histone H4. However, GBD hybrids with SIR2, SIR3, or SIR4 cannot establish silencing. Telomeric silencing, which does not require SIR1 and is normally unstable, is greatly improved by tethering GBD-SIR1 to the telomere. These experiments support a model in which native SIR1 protein is brought to the HM loci by proteins bound to the silencers. Telomeres appear to lack the ability to recruit SIR1, and that is why telomeric silencing is unstable.

Crosses, Genetic↗
PubMed1993

Chromosomal localization and regulation of the longevity determinant genes in a selected strain of Drosophila melanogaster.

A controlled chromosome substitution experiment was performed on a strain (NDC-L) selected for long life to determine if the genes responsible for the extended-longevity phenotype could be localized to any particular chromosome(s). All 27 different possible combinations of the three major chromosomes of Drosophila melanogaster were constructed and longevities were determined on 3875 individual animals of both sexes and analysed. The results are statistically significant and demonstrate that mean longevity is specified primarily by recessive genes on the third chromosome (c3). The extended longevity phenotype (ELP) is only expressed in those lines which are homozygous for the NDC-L type c3. Loci on the first (c1) and second (c2) chromosomes interact, both positively (c1) and negatively (c2), respectively, such that c1 represses c2 which in turn represses c3. The ELP is fully expressed in the mutual presence and mutual absence of c1 and c2. The significance of these results is discussed in the context of broader categories of molecular genetic mechanisms suggested previously to be involved in the modulation of longevity in Drosophila.

Animals↗
PubMed1993

Larval regulation of adult longevity in a genetically-selected long-lived strain of Drosophila.

Our previous work has shown that the major genes involved in the expression of the extended-longevity phenotype are located on the third chromosome. Furthermore, their expression is negatively and positively influenced by chromosomes 2 and 1, respectively. In this report we show that the expression of the extended-longevity phenotype is dependent on the larval environment. A controlled chromosome substitution experiment was carried out using a strain selected for long life (L) and its parent (R) strain. Twenty different combinations of the three major chromosomes were conducted and their longevities were determined under both high (HD) and low (LD) larval density conditions. The extended-longevity phenotype was only expressed under HD conditions. The chromosome interactions were not apparent under LD conditions. Density-shift experiments delineate a critical period for expression of the extended-longevity phenotype, extending from 60 h after egg laying (AEL) to 96 h AEL, during which the developing animal must be exposed to HD conditions if the extended-longevity phenotype is to be expressed. The change from HD to LD conditions is accompanied by statistically significant increases in body weight. The possible role of a dietary restriction phenomenon is examined and the implications of these findings discussed. It is now apparent, however, that the extended-longevity phenotype in Drosophila is a developmental genetic process.

Animals↗
PubMed1993

Doxorubicin cardiomyopathy is associated with a decrease in calcium release channel of the sarcoplasmic reticulum in a chronic rabbit model.

Doxorubicin is a highly effective cancer chemotherapeutic agent that produces a dose-dependent cardiomyopathy that limits its clinical usefulness. Clinical and animal studies of morphological changes during the early stages of doxorubicin-induced cardiomyopathy have suggested that the sarcoplasmic reticulum, the intracellular membrane system responsible for myoplasmic calcium regulation in adult mammalian heart, may be the early target of doxorubicin. To detect changes in the calcium pump protein or the calcium release channel (ryanodine receptor) of the sarcoplasmic reticulum during chronic doxorubicin treatment, rabbits were treated with intravenous doxorubicin (1 mg/kg) twice weekly for 12 to 18 doses. Pair-fed controls received intravenous normal saline. The severity of cardiomyopathy was scored by light and electron microscopy of left ventricular papillary muscles. Developed tension was measured in isolated atrial strips. In subcellular fractions from heart, [3H]ryanodine binding was decreased in doxorubicin-treated rabbits (0.33 +/- 0.03 pmol/mg) compared with control rabbits (0.66 +/- 0.02 pmol/mg; P < 0.0001). The magnitude of the decrease in [3H]ryanodine binding correlated with both the severity of the cardiomyopathy graded by pathology score (light and electron microscopy) and the decrease in developed tension in isolated atrial strips. Bmax for [3H]ryanodine binding and the amount of immunoreactive ryanodine receptor by Western blot analysis using sequence-specific antibody were both decreased, consistent with a decrease in the amount of calcium release channel of sarcoplasmic reticulum in doxorubicin-treated rabbits. In contrast, there was no decrease in the amount or the activity of the calcium pump protein of the sarcoplasmic reticulum in doxorubicin-treated rabbits. Doxorubicin treatment did not decrease [3H]ryanodine binding or the amount of immunoreactive calcium release channel of sarcoplasmic reticulum in skeletal muscle. Since the sarcoplasmic reticulum regulates muscle contraction by the cyclic uptake and release of a large internal calcium pool, altered function of the calcium release channel could lead to the abnormalities of contraction and relaxation observed in the doxorubicin cardiomyopathy.

Animals↗
PubMed1991

Elevated paraquat resistance can be used as a bioassay for longevity in a genetically based long-lived strain of Drosophila.

A long-lived (L) strain of Drosophila melanogaster, derived from a normal-lived (R) strain by artificial selection, has a significantly different adult longevity. Previous work has shown that 1) the two strains age in the same manner, 2) the major genes responsible for much of the L strain's extended longevity are located on the 3rd chromosome, and 3) the extended longevity phenotype is significantly modulated by the larval environment. In this report, we investigate the resistance of the L and R strains to the lethal effects of dietary paraquat. We show that, within the limitations of our described chromosomal and environmental manipulations, the extended longevity phenotype always accompanies the phenotype of elevated paraquat resistance. In addition, reversed selection applied to the L strain results in the simultaneous decrease of both life span and paraquat resistance. Thus, the presence or absence of the latter phenotype may be used as a bioassay for the presence or absence of the extended longevity phenotype, without any necessary implication of causality. Use of this bioassay should greatly speed up the genetic analysis of this system by allowing us to identify long-lived animals at a young age. Finally, we show that the age-related loss of elevated paraquat resistance in both strains precedes all the other age-related functional decrements which we have previously noted in this system.

Animals↗
PubMed1990

Effect of gestational sex steroid exposure on limb development and endochondral ossification in the pregnant C57Bl/6J mouse: I. Medroxyprogesterone acetate.

Although data supporting the teratogenic potential of intrauterine progestin exposure is lacking, concern persists among some individuals within the scientific community that these drugs have the potential for nongenital teratogenesis, especially with regard to limb reduction defects. Our laboratory has been interested in the ontogeny of steroid receptors in the developing embryo and in the role of steroid-receptor interactions in limb development, particularly the process of endochondral ossification. Since limb reduction defects can be produced from abnormal processes that are operative during organogenesis or during midgestation (vascular disruption) we have designed an animal study whereby embryos were exposed to sex steroids throughout organogenesis and fetal development. The present study assesses the effects of medroxyprogesterone acetate (MPA) on intrauterine endochrondral bone development specifically, as well as overall embryo-fetal development. Primagravid C57Bl/6J mice were treated via subdermal pellets which deliver MPA at dosages of 5.0, 50.0, and 500.0 mg/kg/day on gestational days 7 through 19. These doses were 25-, 250-, and 2,500-fold higher on a mg/kg basis than the human dose equivalent (HDE). No increases in nongenital malformations were noted at any evaluated MPA dosage level. At 25 X the HDE, MPA did not influence endochondral bone development as evidenced by a lack of significant effects on assessed bone growth parameters. In the 250- and 2,500-fold HDE dosage groups, MPA was shown to exert an embryotoxic effect inducing 48 and 100% resorptions respectively. Mean embryo weights/litter were significantly reduced by MPA exposure at 250 X the HDE. Intrauterine exposure to 250 X the MPA HDE induced reductions in humeral and femoral diaphyseal length in proportion to a reduction in overall growth. The data demonstrate that MPA, administered at dosages of up to several orders of magnitude in excess of the HDE and which permitted embryo survival, did not induce increases in the frequency of nongenital teratogenesis at any dose or gestational stage. Importantly, limb reduction defects were not noted even in instances where the dosage of MPA induced an inhibition of endochondral bone growth.

Animals↗
PubMed1989

Results of the use of a pure urinary FSH stimulation regime in patients unsuccessfully treated with hMG in an in vitro fertilization program.

63 infertile patients who failed in achieving pregnancy for several cycles with human menopausal gonadotropin stimulation protocol in our in vitro fertilization program were subsequently treated with urinary FSH regime. In 25 of these patients, the leading diagnosis was an elevated basal LH/FSH ratio (i.e. above 2), 29 women had an irreparable tubal factor, 9 patients had endometriosis, and 21 couples suffered from an additional male factor. The FSH stimulation protocol was initiated on day 2 of the cycle, the highest doses were given on the first days and reduced thereafter. Hormone measurements concerning estradiol and LH were started on day 2 of the cycle, ultrasonic evaluation on day 7 of the cycle. After 48 pelviscopic follicle punctures and 39 embryotransfers 12 cases resulted in clinical pregnancies. One case terminated in a miscarriage and 2 pregnancies were of ectopic location.

Adult↗
PubMed1988

Metabolic rates in genetically based long lived strains of Drosophila.

The goal of these experiments was to determine if the increased longevity characteristics of our genetically selected long lived line of Drosophila could be attributed to metabolic differences. The data shows an inverse relationship between life span and temperature for both the long lived (L) and normal (R) strains; however, the higher longevity of the L strain relative to the R strain is not affected by these treatments. Therefore, the genetic factors unique to the L strain do not affect the same processes affected by the temperature treatments. A second set of experiments detected a linear relationship between the MDMR (mean daily metabolic rate) and the ambient adult temperature. However, at each temperature, the MDMR of either strain was statistically equivalent; a finding which demonstrates that an increased life span depends on something other than conservation of calories. A third set of experiments looked at the metabolic efficiency of the two strains and were not able to detect any statistically significant differences. The two strains appear to expend approximately equivalent numbers of calories per day in an approximately equivalent manner. These data are interpreted in the context both of a previously postulated genetic switch mechanism believed responsible for initiating the onset of senescence, and of contemporary reinterpretations of the "rate of living" theory which implicates the essential role of various anti-oxidant defense systems.

Aging↗
PubMed1986

Probenecid induced immune hemolytic anemia.

We report a patient who, while receiving probenecid and colchicine for acute gouty arthritis, developed severe hemolytic anemia in association with a generalized rash. The hemolysis was immune mediated as shown by a positive direct Coombs' test. In vitro hematologic studies showed that a probenecid dependent antibody was present.

Anemia, Hemolytic, Autoimmune↗
PubMed1983

Effects of Mycobacterium bovis (strain BCG) on the interstitial cells of hydronephrotic, contralateral, and normal rabbit kidneys.

Studies were undertaken to determine the effect of viable organisms of Mycobacterium bovis, strain Bacillus Calmette Guerin (BCG), on cell growth characteristics and phagocytic properties of cells from surgically-induced unilaterally hydronephrotic, contralateral, and normal rabbit kidneys. A single intravenous administration of 8 X 10(8) BCG organisms was given at the time of ureteral ligation. Four days after injection, explants were removed from the hydronephrotic, contralateral, and normal kidneys. Two cell types, fibroblasts and mononuclear phagocytes, grew from these explants. BCG caused a marked increase in the rate of growth of cells from the hydronephrotic and contralateral kidneys. There was no measurable effect of BCG on cells from the normal kidney.

Animals↗