Small samples: does size matter?
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Humans↗
Biomedical subjects
A J Anderson
Publications and source records attributed to A J Anderson.
Characterization and expression of the pseudomonas putida bacterioferritin alpha subunit gene.
The root-colonizing pseudomonad Pseudomonas putida (Pp) appears to produce two subunits, alpha and beta, of the iron-binding protein, bacterioferritin. A gene encoding the alpha-bacterioferritin subunit was located adjacent to the major catalase in Pp. The deduced protein sequence of the Pp bfralpha gene had a very high identity with other alpha-subunits, possessing conserved amino acids responsible for ferroxidase activity. The gene also lacked a deduced methionine at residue 52, associated with heme binding in beta-subunits. An antibody generated toward the Escherichia coli (E. coli) multifunctional single subunit bacterioferritin recognized two proteins in the Pp extract, a 22 kDa protein likely to be a beta-subunit and, to a lesser extent, a 23 kDa band. The 23 kDa band was absent in a Pp mutant in which the bfralpha gene was disrupted. Loss of alpha-bacterioferritin stimulated production of fluorescent siderophore. Growth on media and on root surfaces was not impaired by deletion of the alpha-bacterioferritin. Transcription of bfralpha was independent of the catalase gene and was dependent on iron. The transcript levels from bfralpha decreased in iron deficiency experienced during stationary-phase or upon treatment during growth with an iron chelator.
Bacterial Proteins↗
Neurotrophic effects of FPF-1070 (Cerebrolysin) on cultured neurons from chicken embryo dorsal root ganglia, ciliary ganglia, and sympathetic trunks.
We examined the effect of FPF-1070 (Cerebrolysin) on neurite outgrowth in explant cultures of dorsal root ganglia (DRG), sympathetic trunks (ST), and ciliary ganglia (CG) from 10- to 11-day chicken embryos. FPF-1070 significantly promoted neurite outgrowth in DRG and ST neurons at all concentrations examined, in comparison with phosphate buffered saline-treated negative controls; however, this effect on neurite outgrowth was not as significant as that observed for nerve growth factor-treated positive controls on DRG and ST neurons. Additionally, FPF-1070 exhibited an inverted U relationship between concentration and effectiveness in DRG and ST neurons. In contrast, FPF-1070 did not affect neurite outgrowth in CG neurons although ciliary neurotrophic factor-treated positive controls showed striking neurite outgrowth. Our results demonstrate that FPF-1070 has different neurotrophic effects depending on the subpopulation of neurons. This study clarifies a role for neurotrophic activity in the mechanism of action of FPF-1070.
Amino Acids↗
Interactions between flicker thresholds and luminance pedestals.
We investigated the interactions between flicker thresholds and luminance pedestals using threshold versus contrast (TvC) and method of constant stimuli paradigms. High amplitude luminance pedestals were found to elevate flicker thresholds, but low amplitude luminance pedestals were unable to reduce flicker thresholds. Luminance pedestals elevated flicker thresholds more at low temporal frequencies. A simple model based on local light adaptation was able to capture the general form of the TvC functions. Our results suggest that flicker thresholds derived in the presence of a luminance pedestal (luminance-pedestal flicker) may vary from those obtained by modulating about a mean luminance (mean-modulated flicker).
Adaptation, Ocular↗
DNA damage and apoptosis in the aged canine brain: relationship to Abeta deposition in the absence of neuritic pathology.
1. In addition to beta-amyloid (Abeta) deposition and cytoskeletal neuropathology, both the Alzheimer's disease (AD) and Down's syndrome (DS) human brain exhibit marked evidence of DNA damage, however, it is difficult to separate events that occur in conjunction with neurofibrillary pathology versus Abeta pathology in these systems. 2. In contrast, the aged canine brain exhibits the accumulation of Abeta into diffuse deposits similar to those found in early AD and DS in the absence of neurofibrillary pathology. Furthermore, Abeta deposition in canine brain is correlated with cognitive deficits. 3. In order to test the hypothesis that TUNEL labeling for DNA damage in AD is not simply a consequence of agonal artifacts, postmortem artifacts, or neurofibrillary pathology, and may be directly related to Abeta deposition, we examined Abeta immunoreactivity, PHF-1 immunoreactivity, and TUNEL labeling in this animal model. 4. These experiments reveal a relationship between the amount of DNA damage detected by TUNEL labeling and levels of Abeta deposition. Further, in animals with no TUNEL labeling, we detected no Abeta immunoreactivity. 5. These data support the hypothesis that TUNEL labeling in AD ans DS is not a consequence of agonal artifact, postmortem artifact, or tau pathology, and may be directly related to Abeta deposition and perhaps AD pathogenesis.
Aging↗
Effect of stimulus duration in flicker perimetry.
We investigated the relationship between stimulus duration and flicker thresholds when flickering stimuli were presented on luminance pedestals. Mean-modulated flicker thresholds remained constant with changes in stimulus duration. However, flicker presented on a luminance pedestal gave masking at short durations, decaying exponentially to stable thresholds with time. These stable thresholds were elevated when compared with the mean-modulated condition. The lowest threshold in a stimulus onset asynchrony function predicted the threshold obtained from a luminance-pedestal flicker stimulus of the same duration. This suggests that luminance-pedestal flicker thresholds are determined by the most detectable cycle in a multiple cycle stimulus. We find that the 800 ms stimulus duration used in the Medmont M600 perimeter (Medmont, Camberwell, Australia) is suitable to determine stable flicker thresholds.
Adult↗
New millennium bronchodilators for asthma: single-isomer beta agonists.
Racemic beta2 agonists are composed of a 50:50 mixture of R and S isomers. The R isomer exhibits virtually all the bronchodilation, whereas the S isomers are generally considered inert. However, (S)-albuterol was shown to enhance bronchial reactivity to methacholine, eosinophil activation, and histamine-induced influx of fluid, proteins, and neutrophils into the airspaces. Actions such as these may compress the potency and foreshorten the duration of (R)-albuterol. Accordingly, pure (R)-albuterol provides bronchodilation at lower doses than racemate, allowing for fewer beta-adrenergic-mediated side effects. In addition, differential metabolism may allow for the progressive accumulation of (S)-albuterol. This logic is applicable to long-acting beta2 agonists: the therapeutically active (R,R)-formoterol is currently being developed in the United States, and preliminary results suggest rapid improvements in FEV1 with up to 24-hour duration of action. These combined observations with the R isomers of beta2 agonists suggest that potential improvements in therapeutic indices can be achieved with isomerically pure versions of existing racemic drugs.
Adrenergic beta-Agonists↗
The brain's microenvironment, early functional loss, and the conversion to Alzheimer's disease.
We hypothesize that insults in the local microenvironment may cause selective, local neurite degeneration by apoptotic-like mechanisms, leading to the loss of synaptic connectivity observed in the Alzheimer disease brain. This may be an early mechanism of neurodegeneration, potentially contributing to neuronal vulnerability and disease pathogenesis.
Alzheimer Disease↗
Superoxide dismutase activity in Pseudomonas putida affects utilization of sugars and growth on root surfaces.
To investigate the role of superoxide dismutases (SOD) in root colonization and oxidative stress, mutants of Pseudomonas putida lacking manganese-superoxide dismutase (MnSOD) (sodA), iron-superoxide dismutase (FeSOD) (sodB), or both were generated. The sodA sodB mutant did not grow on components washed from bean root surfaces or glucose in minimal medium. The sodB and sodA sodB mutants were more sensitive than wild type to oxidative stress generated within the cell by paraquat treatment. In single inoculation of SOD mutants on bean, only the sodA sodB double mutant was impaired in growth on root surfaces. In mixed inoculations with wild type, populations of the sodA mutant were equal to those of the wild type, but levels of the sodB mutant and, to a great extent, the sodA sodB mutant, were reduced. Confocal microscopy of young bean roots inoculated with green fluorescent protein-tagged cells showed that wild type and SOD single mutants colonized well predominantly at the root tip but that the sodA sodB double mutant grew poorly at the tip. Our results indicate that FeSOD in P. putida is more important than MnSOD in aerobic metabolism and oxidative stress. Inhibition of key metabolic enzymes by increased levels of superoxide anion may cause the impaired growth of SOD mutants in vitro and in planta.
Blotting, Southern↗
Transcriptional regulation by iron of genes encoding iron- and manganese-superoxide dismutases from Pseudomonas putida.
Genes from Pseudomonas putida (Pp), sodA, encoding manganese-superoxide dismutase (MnSOD) and, sodB, iron-superoxide dismutase (FeSOD) were cloned by hybridization with digoxigenin (dig)-labeled PCR products generated from Pp genomic DNA. The sodB gene had a 594 bp open reading frame (ORF), corresponding to 198 amino acids (aa), and a transcript of 880 bases. The sodA gene contained a 609 bp ORF encoding 203 aa and was transcribed as part of a polycistronic operon, consisting of orfY-fumC-orfX-sodA. Pp sodA or sodB genes both restored aerobic growth, growth on paraquat, and growth on minimal medium to an Escherichia coli (Ec) mutant deficient in SOD activity. Paraquat treatment did not enhance mRNA transcription of the sod genes or increase SOD activity in Pp. The Pp sodB gene was highly expressed throughout logarithmic-(log) growth phase and stationary-phase cells grown in medium supplemented with FeCl3, but was down-regulated in iron-deficient conditions, such as in stationary-phase or generated by 2,2'-dipyridyl (DP) treatment. This is the first evidence that iron regulates expression of the sodB gene at the transcriptional level. In contrast, iron-deficient conditions, or addition of MnCl2 to the growth medium, induced transcripts (2.4 kb and 1.2 kb) from the sodA operon. Our results reveal an intricate role of iron in the transcriptional regulation of both Pp sodA and sodB genes.
Cell Division↗
XAFS measurements on zinc chloride aqueous solutions from ambient to supercritical conditions using the diamond anvil cell.
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Journal Article↗
Variant RSH/Smith-Lemli-Opitz syndrome with atypical sterol metabolism.
The RSH/Smith-Lemli-Opitz syndrome (RSH/SLOS) is an autosomal recessive malformation syndrome comprising microcephaly, developmental and growth retardation, characteristic facial anomalies, midline cleft palate, and genital and limb anomalies. Recently, biochemical evidence of an inborn error of cholesterol biosynthesis at the level of 7-dehydrocholesterol (7DHC) reductase was reported in children and adults with RSH/SLOS. We report on two sibs with a variant form of RSH/SLOS whose sterol metabolism in cultured lymphoblasts is abnormal but differs from that of patients with classical RSH/SLOS. The children have relatively mild physical and developmental abnormalities, but a phenotype still consistent with the diagnosis of RSH/SLOS. Their plasma cholesterol levels are only mildly depressed, and they have less markedly increased plasma levels of 7DHC than most patients with classical RSH/SLOS. Cultured lymphoblasts from our patients accumulated 7DHC to the same degree as classical RSH/SLOS lymphoblast when grown with cholesterol-depleted fetal calf serum. However, unlike other RSH/SLOS cells, the increase in cellular 7DHC levels was not suppressed when the cells were grown in the presence of cholesterol from untreated fetal calf serum. The parents' sterol metabolism was also strikingly abnormal in that the levels of 7DHC in their lymphoblasts were markedly elevated compared with those of lymphoblasts from other RSH/SLOS parents. Our findings suggest that these mildly affected RSH/SLOS sibs may have a genetic disorder of sterol metabolism that is related to but biochemically different from classical RSH/SLOS, possibly one affecting intracellular transport of sterols.
Cholesterol↗
Relation of lipoprotein subclasses as measured by proton nuclear magnetic resonance spectroscopy to coronary artery disease.
Although each of the major lipoprotein fractions is composed of various subclasses that may differ in atherogenicity, the importance of this heterogeneity has been difficult to ascertain owing to the labor-intensive nature of subclass measurement methods. We have recently developed a procedure, using proton nuclear magnetic resonance (NMR) spectroscopy, to simultaneously quantify levels of subclasses of very low density (VLDL), low density (LDL), and high density (HDL) lipoproteins; subclass distributions determined with this method agree well with those derived by gradient gel electrophoresis. The objective of the current study of 158 men was to examine whether NMR-derived lipoprotein subclass levels improve the prediction of arteriographically documented coronary artery disease (CAD) when levels of lipids and lipoproteins are known. We found that a global measure of CAD severity was positively associated with levels of large VLDL and small HDL particles and inversely associated with intermediate size HDL particles; these associations were independent of age and standard lipid measurements. At comparable lipid and lipoprotein levels, for example, men with relatively high (higher than the median) levels of either small HDL or large VLDL particles were three to four times more likely to have extensive CAD than were the other men; the 27 men with high levels of both large VLDL and small HDL were 15 times more likely to have extensive CAD than were men with low levels. In contrast, adjustment for levels of triglycerides or HDL cholesterol greatly reduced the relation of small LDL particles to CAD. These findings suggest that large VLDL and small HDL particles may play important roles in the development of occlusive disease and that their measurement, which is not possible with routine lipid testing, may lead to more accurate risk assessment.
Adult↗
Comparison of outcome of implantable cardioverter defibrillator implantation in patients with severe versus moderately severe left ventricular dysfunction secondary to atherosclerotic coronary artery disease.
This study was undertaken to assess the feasibility and clinical outcome of implantable cardioverter-defibrillators (ICDs) among patients with coronary artery disease and left ventricular ejection fraction (LVEF) of <20%. The morbidity, mortality, and the long-term survival of 117 patients with LVEF of <20% (group 1) were compared with 321 patients with LVEF of 20% to 40% (group 2). Mortality in the first 30 days after ICD implantation was 0% for group 1 and 0.6% in group 2. Actuarial survival (all cause) at the end of 2, 4, and 5 years were 83%, 70%, and 62%, respectively, in group 1 and 90%, 80%, and 71% in group 2 (p = 0.05). Fifty-five patients (47%) in group 1 and 126 patients (39%) in group 2 received appropriate shocks during follow up. Among the patients in group 1, the overall survival at 2 years after an appropriate shock from an ICD was 92% for patients <60 years of age, 77% for patients ages 60 to 69, and 53% for patients >70 years old. Although the overall survival of patients in group 1 was slightly lower compared with those in group 2, in a multivariate analysis, the EF was not an independent predictor of poor survival. The ICD can be implanted with acceptable operative morbidity and mortality in selected patients with LVEF of <20%.
Aged↗
Identification of adjacent genes encoding the major catalase and a bacterioferritin from the plant-beneficial bacterium Pseudomonas putida.
The catA gene, encoding the major CatA from a root-colonizing isolate Pseudomonas putida (Pp), was cloned by complementation into a catalase (Cat)-deficient Escherichia coli (Ec) strain UM2. The ORF for catA consisted of 479 aa with a higher degree of identity with typical Cat from eukaryotes than prokaryotes. Chromosomal homologous exchange with a mutant gene bearing an insertion of a luxAB-npt cassette into the SfiI site of catA generated a CatA-deficient Pp isolate. This mutant and another mutant, J1M, derived by EMS mutagenesis, were highly sensitive to hydrogen peroxide. CatA activity and resistance to hydrogen peroxide were restored in both mutants by catA. Adjacent to the 3' end of catA was a potential ORF of 462 nt that had high identitity with other bfr genes that encode iron-storage proteins. Northern analysis of the bfr gene from Pp revealed a transcript of approximately 500 nt. CatA and bfr probes hybridized to the same size restriction fragments in genomic DNAs from other root-colonizing and plant pathogenic pseudomonads. Thus, the genes for an iron-storage protein and the heme-containing Cat appear to be conserved in adjacent loci in certain pseudomonads.
Amino Acid Sequence↗
Efficacy of the implantable cardioverter-defibrillator in the elderly.
OBJECTIVES: We sought to assess the effect of advanced age on the outcome of patients with an implantable cardioverter-defibrillator (ICD). BACKGROUND: ICDs are effective in preventing sudden cardiac death in susceptible patients, but their beneficial effect on survival is attenuated by the high rate of nonsudden cardiac death in those treated. Although advanced age is an important variable in determining cardiovascular mortality, its impact on the outcome of patients with an ICD has been inadequately studied. METHODS: We performed multivariate analysis of a data base consisting of 769 consecutive patients with an ICD. Seventy-four patients > or = 75 years old at ICD implantation (Group 1) were compared with the remaining 695 patients (Group 2). RESULTS: The two groups were similar in clinical presentation, left ventricular function and gender distribution. The mean follow-up time was 29 and 42 months, respectively, for patients in Group 1 and Group 2. Actuarial survival at 4 years was 57% in Group 1 versus 78% in Group 2 (p = 0.0001). This difference was primarily due to a higher rate of nonsudden cardiac death in Group 1. On multivariate analysis, age > or = 75 years, New York Heart Association functional class III, left ventricular ejection fraction < 30% and appropriate shocks during follow-up were independently associated with increased mortality (odds ratio 3.56, 1.8, 1.6 and 1.39, respectively). CONCLUSIONS: Among patients with similar functional class and ejection fraction, the mortality risk is increased threefold in those > or = 75 years old at the time of ICD implantation. Extrapolation of results from younger patients is likely to overestimate ICD benefit in the elderly.
Age Factors↗
The effects of insulin/IGF-I on glucose and leucine metabolism in the redclaw crayfish (Cherax quadricarinatus).
In recent years, invertebrate peptides have been identified which share substantial homologies with vertebrate insulin and insulin-like growth factors (IGFs), indicating a high degree of conservation of insulin/IGF systems through animal evolution. In a previous study, we provided evidence for the presence of IGF-I-like peptides in the redclaw (Cherax quadricarinatus), a species of freshwater crayfish endemic to northern Australia river systems which has attained support as a culture species. The general aim of the current study was to elucidate the functional significance of IGF-I-like peptides in this species by examining the effects of mammalian IGF-I on glucose and leucine metabolism. Juvenile redclaw were injected with a single dose of purified human insulin, recombinant human (rh) IGF-I, or Des-1-3-IGF-I. Glucose levels in redclaw tissues were then determined over an 8-hr period using enzymatic approaches. It was shown that injection of rhIGF-I induced an acute increase in free glucose content in hepatopancreas while Des-1-3-IGF-I and insulin raised free glucose levels in abdominal muscle. Radiolabel tracer approaches also demonstrated that injection of rhIGF-I increased glycogen synthesis in abdominal muscle and elevated the incorporation of leucine into protein in both abdominal muscle and hepatopancreas. Taken together, the findings of this study suggest that IGF-I-like peptides are biologically active in this species and may be of significance to metabolic and growth-related processes.
Animals↗
Seedborne fungal contamination: consequences in space-grown wheat.
Plants grown in microgravity are subject to many environmental stresses that may promote microbial growth and result in disease symptoms. Wheat (cv. Super Dwarf) recovered from an 8-day mission aboard a NASA (National Aeronautics and Space Administration) space shuttle showed disease symptoms, including girdling of leaf sheaths and chlorosis and necrosis of leaf and root tissues. A Neotyphodium species was isolated from the seed and leaf sheaths of symptomatic wheat used in the spaceflight mission. Certain isozymes of a peroxidase unique to extracts from the microgravity-grown plants were observed in extracts from earth-grown Neotyphodium-infected plants but were not present in noninfected wheat. The endophytic fungus was eliminated from the wheat seed by prolonged heat treatment at 50 degrees C followed by washes with water at 50 degrees C. Plants from wheat seed infected with the Neotyphodium endophyte were symptomless when grown under greenhouse conditions, whereas symptoms appeared after only 4 days of growth in closed containers. Disease spread from an infected plant to noninfected plants in closed containers. Dispersion via spores was found on asymptomatic plants at distances of 7 to 18 cm from infected plants. The size and shape of the conidia, mycelia, and phialide-bearing structures and the ability to grow rapidly on carbohydrates, especially xylose, resembled the characteristics of N. chilense, which is pathogenic on orchard grass, Doctylis glomerati. The Neotyphodium wheat isolate caused disease symptoms on other cereals (wheat cv. Malcolm, orchard grass, barley, and maize) grown in closed containers.
Edible Grain↗