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Biomedical subjects

K A Johnson

Publications and source records attributed to K A Johnson.

At least 109 records · Page 6Linked to original sources
PubMed1998

Preclinical prediction of Alzheimer's disease using SPECT.

BACKGROUND: Regional cerebral perfusion measured by single photon emission computed tomography (SPECT) was examined as a preclinical predictor of the development of Alzheimer's disease (AD). METHODS: Singular value decomposition was used to produce 20 SPECT factors (known as vectors) (n=152). Vector scores were then computed for four groups (n=136), differing in cognitive status: Group 1--normal controls at both baseline and follow-up; Group 2--subjects with "questionable" AD at both baseline and follow-up; Group 3--subjects with questionable AD at baseline who converted to AD on follow-up (Converters); Group 4--subjects with AD at baseline. All SPECT data in the analyses were gathered at baseline. RESULTS: The four groups could be distinguished on the basis of their baseline SPECT data (p < or = 0.00005; hit rate=83%). Regional decreases in perfusion were most prominent among Converters in the hippocampal-amygdaloid complex, the posterior cingulate, the anterior thalamus, and the anterior cingulate. Inclusion of apolipoprotein E status did not significantly improve the discrimination. CONCLUSIONS: SPECT data gathered and analyzed in this manner may be useful as one aspect of the preclinical prediction of AD. Three of the four brain regions important for discriminating Converters from normal controls involve a distributed brain network pertaining to memory, suggesting that this network may be selectively affected in the earliest stages of AD.

Aged↗
PubMed1998

The axonemal microtubules of the Chlamydomonas flagellum differ in tubulin isoform content.

Little is known of the molecular basis for the diversity of microtubule structure and function found within the eukaryotic flagellum. Antibodies that discriminate between tyrosinated alpha tubulin and post-translationally detyrosinated alpha tubulin were used to localize these complementary tubulin isoforms in flagella of the single-celled green alga Chlamydomonas reinhardtii. Immunofluorescence analysis of intact axonemes detected both isoforms along most of the lengths of flagella; however, each had a short distal zone rich in tyrosinated tubulin. Localizations on splayed axonemes revealed that the microtubules of the central-pair apparatus were rich in tyrosinated tubulin, while outer doublets contained a mixture of both isoforms. Immunoelectron analysis of individual outer doublets revealed that while tyrosinated tubulin was present in both A and B tubules, detyrosinated tubulin was largely confined to the wall of the B hemi-tubules. The absence of detyrosinated tubulin from the A tubules of the outer doublets and the microtubules of the central pair, both of which extend past the B hemi-tubules of the outer doublets in the flagellar tip, explained the appearance of a tyrosinated tubulin-rich distal zone on intact axonemes. Localizations performed on cells regenerating flagella revealed that flagellar assembly used tyrosinated tubulin; detyrosination of the B tubule occurred during later stages of regeneration, well after microtubule polymerization. The developmental timing of detyrosination, which occurs over a period during which the regrowing flagella begin to beat more effectively, suggests that post-translational modification of the B tubule surface may enhance dynein/B tubule interactions that power flagellar beating.

Animals↗
PubMed1998

Alcohol dependence in adult children of alcoholics: longitudinal evidence of early risk.

This study investigates familial alcoholism effects and the comparative probability of risk for alcohol dependence in adult children of alcoholics (ACAs) with a control group of non-ACAs. A cohort of 12,686 young adults from the National Longitudinal Survey of Youth (NLSY) is examined over a five-year period and conventional and lineal intergenerational models of alcoholism transmission are assessed. The results of multivariate logistic regression analyses indicate that the risk is relatively greater for male ACAs; sons of alcoholics drink significantly more heavily, experience problems earlier, and develop alcohol dependence more extensively than female ACAs or non-ACAs of either gender. The extent of dependence found in subjects with a lineal history of alcoholism on the father's side of the family, as well as heavy drinking, cigarette smoking and drinking onset in adolescence should be considered as critical predisposing factors of high risk for dependence at later ages. These observations corroborate clinical studies and support a growing body of biopsychosocial research literature.

Adult↗
PubMed1998

Effect of level and source of dietary selenium on concentrations of thyroid hormones and immunoglobulins in beef cows and calves.

Our objective was to determine the effect of level and chemical form of dietary selenium on productivity of beef cows, concentrations of triiodothyronine (T3), and thyroxine (T4) in plasma, and immunoglobulins (IgG and IgM) in plasma and colostrum of cows. Pregnant cows (n = 60) were randomly allocated among four dietary treatments of 20, 60, or 120 ppm Se as selenite and 60 ppm as selenomethionine from selenized yeast (SeY) in salts offered free-choice. Treatments began 90 d prepartum and continued through the second parturition. Treatments did not affect the final body weights of cows or birth weights or weaning weights of calves. At parturition, cows given salt with 20 ppm Se as selenite had lower (P < . 05) concentrations of Se in blood than cows with access to higher-Se salts. Treatments affected (P < .01) the concentration of T3 and the ratio of T3:T4 in plasma of cows. The concentration of T3 in plasma of cows with access to salt with 20 ppm Se was 14% lower than that in cows supplemented with 60 ppm Se as selenite or SeY. Plasma IgG in cows and calves, colostrum, and Se concentrations in colostrum, casein, and whey were lowest (P < .01) for cows given salt with only 20 ppm Se. Thus, salts with concentrations of 60 and 120 ppm Se improved measures of Se status in cows and calves. Consideration should be given to the concentrations of T3 and IgG when determining the nutritional requirements for Se in cattle.

Animal Feed↗
PubMed1998

Use of singular value decomposition to characterize age and gender differences in SPECT cerebral perfusion.

UNLABELLED: The main objective of this study was to characterize changes in brain perfusion associated with normal aging and gender. METHODS: Perfusion SPECT images using 99mTc-hexamethyl propyleneamine oxime (HMPAO) were obtained from 152 healthy subjects (67 men, 85 women) aged 50-92 yr. An automated method was developed to objectively assess image data from a large number of brain regions. Image data were reduced with singular value decomposition (SVD), which produced 20 eigenvectors capturing 97.05% of the total information content of 4320 regions from each subject. Subjects were scored individually on each vector. RESULTS: Multivariate analyses demonstrated that there were no significant differences in whole-brain HMPAO uptake with age, but age-related regional declines were seen in lateral ventricular regions. Women had higher HMPAO uptake than men in estimates of global perfusion and regional perfusion in the midcingulate/corpus callosum, inferior temporal and inferior parietal areas. CONCLUSION: These discriminations demonstrate that singular value deomposition of SPECT data may be used to assess differences in perfusion patterns between groups of subjects. They replicate several previous findings, both with respect to age-related changes in perfusion and with respect to gender differences. In addition, they identify a previously unreported gender difference in biparietal regions.

Aged↗
PubMed1997

RNA secondary structure switching during DNA synthesis catalyzed by HIV-1 reverse transcriptase.

Changes in RNA secondary structure have been found to play important roles in translational regulation, protein synthesis, and mRNA splicing. In studies utilizing a 66 nucleotide RNA template with a stable hairpin structure, we have examined the effects of RNA secondary structure on HIV-1 reverse transcriptase activity. We identify several pause sites in the stem of the hairpin and show that these pause sites are correlated with the free energy of melting the next base pair in the stem. We also identify a pause site appearing in the loop of the hairpin and show that this is due to the rapid formation of a new hairpin structure occurring during the progress of DNA polymerization through the hairpin. The rapid change in RNA secondary structure to form the new hairpin selectively destabilizes the major hairpin and thereby accelerates the rate at which reverse transcriptase reads through RNA secondary structure.

Base Sequence↗
PubMed1997

A hexameric helicase encircles one DNA strand and excludes the other during DNA unwinding.

The bacteriophage T7 DNA helicase/primase (gene 4 protein) is a ring-like hexamer that encircles ssDNA and requires forked DNA to catalyze DNA unwinding. We report that optimal rates of unwinding of forked DNA require ssDNA tails of 55 nucleotides on the 5'-to-3' strand and 15 nucleotides on the 3'-to-5' strand. Surprisingly, streptavidin bound to a biotinylated 3'-end fully substitutes for the 3'-to-5' ssDNA tail. This suggests that excluding the 3'-to-5' DNA strand from the center of the helicase is an essential aspect of the mechanism of hexameric helicase-catalyzed DNA unwinding. We also report that streptavidin bound to a biotinylated dT within the 5'-to-3' strand of the duplexed region abolishes DNA unwinding; whereas, streptavidin bound to a biotinylated dT within the duplexed region of the other strand has no effect. These results unambiguously demonstrate that the T7 gene 4 protein is a 5'-to-3' helicase and imply that during DNA unwinding the 5'-to-3' strand transverses the center of the ring while the 3'-to-5' strand is excluded from the center of the ring. Implications for collisions between a helicase and other protein-DNA complexes are discussed.

Bacteriophage T7↗
PubMed1997

Increase of 5HT and VIP immunoreactivity within the hamster (Mesocricetus auratus) SCN during chronic MAOI treatment.

The effects of chronic treatment with the monoamine oxidase inhibitor (MAOI), clorgyline (CLG; 2 mg/kg per day) on serotonin (5HT) and vasoactive intestinal peptide (VIP) immunoreactivity (IR) within the hamster suprachiasmatic nucleus (SCN) were examined. Optical densities of 5HT IR and VIP IR were increased by MAOI treatment. VIP IR was increased in both the ventrolateral and dorsal regions of the SCN, suggesting that VIP content was increased within both perikarya and terminals of VIP neurons. The results suggest that previously described effects of MAOIs on the mammalian circadian system may be mediated in part, by their effects on serotonergic input to VIP neurons within the SCN.

Animals↗
PubMed1997

Effect of RNA secondary structure on the kinetics of DNA synthesis catalyzed by HIV-1 reverse transcriptase.

The effect of RNA secondary structure on the kinetics of DNA synthesis catalyzed by HIV-1 RT was determined using a 66 nucleotide RNA template containing a stable 12 base pair hairpin structure. Prior to reaching the hairpin structure, the primer elongation by RT was fast and the kinetics of polymerization was not affected by the presence of the secondary structure. Once within the regions of template secondary structure, polymerization was much slower and RT paused at five distinct sites [Suo, Z., & Johnson, K. A. (1997) Biochemistry (manuscript submitted for publication)]. Kinetic analysis of single nucleotide incorporation at the pause sites showed polymerization occurred by both a fast phase (54-76 s-1) and a slow phase (0.07-0.4 s-1) during a single binding event. The biphasic kinetics suggests that the DNA substrates are initially bound in both productive and nonproductive states at the polymerase site of RT. The nonproductively bound DNA is slowly converted into a productive state without dissociation from the enzyme. At the pause sites, the enzyme amplitudes of the fast phase are small (4.0-15%) while the amplitudes of the slow phase are large (11-40%). In contrast, only the reaction at the fast phase was observed at the nonpause sites and the enzyme amplitudes were large (63-66%) although the nucleotide incorporation rates (62-78 s-1) are similar to the fast phase rates at the pause sites. These indicate that DNA substrates were bound predominantly nonproductively at pause sites and productively at nonpause sites. However, the overall binding affinity of DNA substrates was measured by the nitrocellulose-DEAE double filter binding assay, binding affinity at both pause sites and nonpause sites was similar (9-38 nM). This indicates that substrates are bound tightly at the large binding cleft of HIV-1, although they may not be productively bound at the polymerase active site. These results and those reported elsewhere [Suo, Z., & Johnson, K. A. (1997) Biochemistry (manuscript submitted for publication)] are consistent with a model in which, at pause sites, HIV-1 RT remains bound to DNA substrates waiting for the melting of the next stem base pair of template secondary structure. Upon melting of the stem base pair, polymerization to fill the open template site is fast and largely irreversible, allowing RT to read through the stable hairpin structures.

Base Sequence↗
PubMed1997

Effect of RNA secondary structure on RNA cleavage catalyzed by HIV-1 reverse transcriptase.

Using a synthetic 66 nucleotide RNA template containing a stable hairpin structure derived from the HIV-1 genome, six predominant RNA cleavage products are found during DNA synthesis catalyzed by HIV-1 RT. These major RNA cleavage sites correlate well with the pause sites seen during primer elongation [Suo, Z., & Johnson, K. A. (1997) Biochemistry (manuscript submitted for publication)]. Thus, the RNase H and polymerase activities of RT are coupled as RT reads through the RNA secondary structure. The distance between the two active sites of HIV-1 RT is 19-20 base pairs of DNA/RNA heteroduplex when the next template base is not paired. The heteroduplex region was enlarged by 2-3 base pairs once RT encounters the template hairpin. A model for this change is presented. At the pause sites, the burst amplitudes of RNA cleavage are larger than the corresponding reaction amplitudes of next nucleotide incorporation at the polymerase site. Measurement of the steady state rates of RNA cleavage confirms that all substrates dissociate slowly from RT. These results suggest that while substrates are bound nonproductively at the polymerase site, they are still bound productively at the RNase H active site of RT. Characterization of an RNase H-deficient RT mutant (D443N) shows that RNase H activity is not critical for RT to read through the RNA secondary structure. HIV-1 nucleocapsid does not increase the processivity of HIV-1 RT but inhibits DNA elongation by blocking the binding of RT to DNA substrates.

Base Sequence↗
PubMed1997

H. M.'s medial temporal lobe lesion: findings from magnetic resonance imaging.

Although neuropsychological studies of the amnesic patient H. M. provide compelling evidence that normal memory function depends on the medial temporal lobe, the full extent of his surgical resection has not been elucidated. We conducted magnetic resonance imaging studies to specify precisely the extent of his bilateral resection and to document any other brain abnormalities. The MRI studies indicated that the lesion was bilaterally symmetrical and included the medial temporal polar cortex, most of the amygdaloid complex, most or all of the entorhinal cortex, and approximately half of the rostrocaudal extent of the intraventricular portion of the hippocampal formation (dentate gyrus, hippocampus, and subicular complex). The collateral sulcus was visible throughout much of the temporal lobe, indicating that portions of the ventral perirhinal cortex, located on the banks of the sulcus, were spared; the parahippocampal cortex (areas TF and TH) was largely intact. The rostrocaudal extent of the ablation was approximately 5.4 cm (left) and 5.1 cm (right). The caudal 2 cm, approximately, of the hippocampus body (normal length, approximately 4 cm) was intact, although atrophic. The temporal stem was intact. Outside the temporal lobes, the cerebellum demonstrated marked atrophy, and the mammillary nuclei were shrunken. The lateral temporal, frontal, parietal, and occipital lobe cortices appeared normal for age 66 years. The mediodorsal thalamic nuclei showed no obvious radiological changes. These findings reinforce the view that lesions of the hippocampal formation and adjacent cortical structures can produce global and enduring amnesia and can exacerbate amnesia beyond that seen after more selective hippocampal lesions.

Aged↗
PubMed1997

Repair of propanodeoxyguanosine by nucleotide excision repair in vivo and in vitro.

Repair of the exocyclic DNA adduct propanodeoxyguanosine (PdG) was assessed in both in vivo and in vitro assays. PdG was site-specifically incorporated at position 6256 of M13MB102 DNA, and the adducted viral genome was electroporated into repair-proficient and repair-deficient Escherichia coli strains. Comparable frequencies of PdG --> T and PdG --> A mutations at position 6256 were detected following replication of the adducted genomes in wild-type E. coli strains. A 4-fold increase in the frequencies of transversions and transitions was observed in E. coli strains deficient in Uvr(A)BC-dependent nucleotide excision repair. A similar increase in the replication of the adduct containing strand was observed in the repair-deficient strains. No change in the frequency of targeted mutations was observed in strains deficient in one or both of the genes coding for 3-methyladenine glycosylase. Incubation of purified E. coli Uvr(A)BC proteins with a duplex 156-mer containing a single PdG adduct resulted in removal of a 12-base oligonucleotide containing the adduct. Incubation of the same adducted duplex with Chinese hamster ovary cell-free extracts also resulted in removal of the adduct. PdG was a better substrate for repair by the mammalian nucleotide excision repair complex than the bacterial repair complex and was approximately equal to a thymine-thymine dimer as a substrate for the former. The results of these in vivo and in vitro experiments indicate that PdG, a homolog of several endogenously produced DNA adducts, is repaired by the nucleotide excision repair pathway.

Animals↗
PubMed1997

Characterization of the mouse gene encoding phospholipid transfer protein.

The mouse gene encoding phospholipid transfer protein (PLTP) was cloned from 129/SvJ lambdaFIX(R) II library and characterized for the first time. It is comprised of 16 exons separated by 15 introns. Its gene organization strikingly resembles that encoding the human PLTP; the exon-intron junctions in these two genes are completely conserved. Sequencing analysis reveals that the putative promoter of mouse PLTP gene consists of a TATA-box, a high GC region, and several consensus sequences for the binding of transcription factors. Within the first 200 bp of the 5'-flanking region, the mouse and human PLTP genes share 81.1% identity of nt sequences and contain the consensus sequences for the transcription factors AP-2 and Sp1 at the same locations.

Animals↗
PubMed1997

Comparative modeling of the three-dimensional structure of the calmodulin-related TCH2 protein from Arabidopsis.

Plants adapt to various stresses by developmental alterations that render them less easily damaged. Expression of the TCH2 gene of Arabidopsis is strongly induced by stimuli such as touch and wind. The gene product, TCH2, belongs to the calmodulin (CaM) family of proteins and contains four highly conserved Ca(2+)-binding EF-hands. We describe here the structure of TCH2 in the fully Ca(2+)-saturated form, constructed using comparative molecular modeling, based on the x-ray structure of paramecium CaM. Like known CaMs, the overall structure consists of two globular domains separated by a linker helix. However, the linker region has added flexibility due to the presence of 5 glycines within a span of 6 residues. In addition, TCH2 is enriched in Lys and Arg residues relative to other CaMs, suggesting a preference for targets which are more negatively charged. Finally, a pair of Cys residues in the C-terminal domain, Cys126 and Cys131, are sufficiently close in space to form a disulfide bridge. These predictions serve to direct future biochemical and structural studies with the overall aim of understanding the role of TCH2 in the cellular response of Arabidopsis to environmental stimuli.

Amino Acid Sequence↗
PubMed1997

Plant responses to environmental stress: regulation and functions of the Arabidopsis TCH genes.

Expression of the Arabidopsis TCH genes is markedly upregulated in response to a variety of environmental stimuli including the seemingly innocuous stimulus of touch. Understanding the mechanism(s) and factors that control TCH gene regulation will shed light on the signaling pathways that enable plants to respond to environmental conditions. The TCH proteins include calmodulin, calmodulin-related proteins and a xyloglucan endotransglycosylase. Expression analyses and localization of protein accumulation indicates that the potential sites of TCH protein function include expanding cells and tissues under mechanical strain. We hypothesize that at least a subset of the TCH proteins may collaborate in cell wall biogenesis.

Arabidopsis↗
PubMed1997

Resorbable and non-resorbable augmentation devices for tenorrhaphy of xenografts in extensor tendon deficits: 12 week study.

Resorbable (poly-L-lactide) and non-resorbable (polyethylene terephathalate) tendon augmentation devices (TAD) in conjunction with a pericardial adhesion barrier, were designed to strengthen tenorrhaphies and were evaluated in an ovine extensor tendon deficit model in a short term study. Fifteen centimetres of tendon were resected and replaced with kangaroo tail tendon xenografts that had been cross-linked with 0.075% glutaraldehyde (GA) at 4 degrees C for one or seven days. Compared with tenorrhaphies performed with Kessler sutures alone, both types of TAD were more effective at preventing tenorrhaphy dehiscence, and thus maintaining tendon function. Furthermore, tensile strength of TAD tenorrhaphies increased significantly between zero and twelve weeks. For xenografts cross-linked in GA for one day, the tensile strength of tenorrhaphies with the resorbable TAD rose from 38 +/- 9 N at time zero, to 116 +/- 46 N at twelve weeks, while non-resorbable TAD tenorrhaphy strength at time zero was 42 +/- 16 N and 99 +/- 27 N at twelve weeks. For xenografts cross-linked with GA for seven days, similar increases in tensile strength of tenorrhaphies, with the two types of TAD were found. As there was no significant difference in mechanical performance or tissue response between the two TAD types in the first 12 weeks, use of the resorbable poly-L-lactide device may be advantageous clinically. Tensile strengths of midsections of the tendon xenograft cross-linked for 7 days was not significantly diminished 12 weeks after implantation and these xenografts were partially remodelled around the periphery. However, the tensile strength of xenografts cross-linked for one day declined significantly between time zero (319 +/- 80 N) and twelve weeks (239 +/- 92 N), suggesting that this degree of cross-linking was inadequate for maintenance of mechanical strength. Evaluation of the performance of tenorrhaphy augmentation devices with xenografts, over a longer implantation period, is required to further understand their usefulness for reconstruction of traumatic tendon injuries.

Animals↗
PubMed1997

Evaluation of a novel assay of potential toxicity/neurotoxicity of carpet emissions (VOCs) in mice.

A private testing laboratory utilizing the whole-body plethysmograph/head-only exposure apparatus outlined in the respiratory irritation assay ASTM E981-84, along with a novel exposure regimen, has reported neurotoxic effects and mortality in mice exposed to relatively low levels of volatile organic compounds (VOCs) emitted from a number of consumer products. This methodology was evaluated by exposing groups of mice, including unrestrained and sham-treated animals, to VOCs generated from a sample of carpet reported to be neurotoxic using the modified assay. General toxicological (haematological measurements, organ weights, gross pathology, histopathology) and specific neurotoxicity (functional observations, body temperature, histopathology of nervous tissues) parameters were evaluated. No effects related to exposure to carpet VOCs were observed in the mice. However, despite careful handling, a number of effects were observed which were attributed to the repeated restraint of mice in the ASTM E981 apparatus. These included a number of minor physical injuries, decreased body weights, altered thymus weights, compression damage to the liver and haemorrhage of the pituitary gland. It was concluded that the modification of the original ASTM E981 methodology may result in physical injuries and stress which may significantly affect any evaluation of toxicity and neurotoxicity in treated animals and result in inaccurate conclusions.

Air Pollutants↗