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Biomedical subjects

G G Brown

Publications and source records attributed to G G Brown.

At least 73 records · Page 4Linked to original sources

In organello transcription in maize mitochondria and its sensitivity to inhibitors of RNA synthesis.

Purified mitochondrial preparations from etiolated maize shoots support the incorporation of radioactivity from labeled UTP into RNA. The incorporation is linear with time for up to 2 hours, shows Michaelis-Menton kinetics with respect to the concentration of the labeled substrate, UTP, and has salt and pH optima which are different than those previously reported for RNA synthesis by isolated chloroplasts. When a crude mitochondrial preparation is subjected to isopycnic sucrose gradient centrifugation, the bulk of the RNA synthetic activity co-sediments with mitochondrial marker enzymes and with the mitochondrial 26S and 18S rRNAs. Maize mitochondrial RNA synthesis is prevented by actinomycin D and ethidium bromide but unaffected by alpha-amanitin. It is strongly inhibited by rifampicin at concentrations which have no effect on nuclear and chloroplast RNA synthesis, but only moderately inhibited by rifampicin at concentrations which completely inhibit bacterial RNA synthesis. The optimization, cell fractionation, and inhibitor data all suggest that contaminating organelles and bacteria do not contribute appreciably to the RNA synthesis in purified mitochondrial preparations.

Journal Article↗

Structural conservation and variation in the D-loop-containing region of vertebrate mitochondrial DNA.

The nucleotide sequences of the D-loop-containing regions of three rat mitochondrial DNAs (mtDNAs), two from the species Rattus norvegicus and one from R. rattus, were determined. Comparisons made among these sequences and with the mouse sequence showed that, on the basis of both base composition and frequency of nucleotide alterations, three domains could be defined within the D-loop-containing region: a central conserved segment, poor in L-strand adenine, flanked by two divergent, adenine-rich regions. Deletions and insertions were found to occur at an unexpectedly high frequency in these sequences and the conserved sequence block called CSB-1 was found not to be intact in the R. rattus sequence. Although in comparisons of more distantly related mtDNAs the D-loop region is the most divergent on the molecule, it does not diverge more than typical protein genes between R. norvegicus and R. rattus, and its central conserved domain appears to be one of the molecule's most conserved regions. The most variable domain borders the tRNAPhe gene and contains the L and H-strand promoters and the 5' terminus for H-strand DNA synthesis. Within this region we have found sequences in all the mtDNAs we have examined, including those of human, two artiodactyls and Xenopus, that are capable of folding into cloverleaf structures. In the other divergent domain of the same mtDNAs, we find sequences capable of assuming similar secondary structural configurations at or near the sites for the termination of D-loop DNA synthesis. The evolutionary preservation of the potential to form such structures despite the high primary-structural divergence of the regions they occur in, suggests the structures are of principal importance for some processes occurring in the D-loop-containing region.

Animals↗

Autonomously replicating RNA in mitochondria of maize plants with S-type cytoplasm.

Mitochondria isolated from maize plants with S-type male-sterile cytoplasms are capable of synthesizing four species of RNA at concentrations of actinomycin D that eliminate all DNA-directed RNA synthesis. No RNA synthesis occurs under the same conditions with mitochondria from plants possessing normal (N) cytoplasm or with other subcellular fractions from plants with S cytoplasm. The actinomycin D-resistant RNA synthesis occurs within the mitochondria since the labeling of these species is unaffected by inclusion of RNase in the incubation medium and since they become completely sensitive to RNase upon lysis of the mitochondria with low concentrations of Triton X-100. Two of the actinomycin D-resistant products are double stranded. These are 2850 and 900 base pairs in length, whereas the remaining two are 2150 and 850 bases. The synthesis of all four RNAs occurs in at least five different accessions of S cytoplasm, suggesting it is a general feature of S mitochondria. The double-stranded RNAs show homology to single-stranded S mitochondrial RNA but not to N mitochondrial RNA. Our observations indicate that the replication of these RNAs occurs independently of mtDNA and that they thus represent a novel type of inheritable element in organelles, an RNA plasmid.

Journal Article↗

Construction of matched verbal and design continuous paired associate tests.

We developed verbal and figural continuous paired associate tests (CPAT) that are matched on internal consistency and on the distributional properties of mean item difficulty, variance, and skewness. The Verbal CPAT is composed of high frequency nouns, verbs, and adjectives. Selection of the designs was guided by literature on the effects of right-hemisphere damage on memory for designs. One hundred subjects were given 80 items from the verbal CPAT and 80 items from the designs CPAT. Twenty subjects at these five ranges of Wechsler Memory Quotient were evaluated: less than or equal to 79, 80-89, 90-99, 100-109, greater than or equal to 109. The final 40-item version of the Verbal CPAT had a mean of 26.72, a SD of 7.03, and an alpha of .859. The Designs CPAT had a mean of 26.42, a SD of 7.75, and an alpha of .886. Neither test was significantly skewed. These matched tests should be useful in making inferences about differences between verbal and figural memory that are not confounded with the problem of differential test sensitivity.

Adolescent↗

Analysis of covariance as a remedy for demographic mismatch of research subject groups: some sobering simulations.

Analysis of Covariance (ANCOVA) is often used in neuropsychological studies to effect ex-post-facto adjustment of performance variables amongst groups of subjects mismatched on some relevant demographic variable. This paper reviews some of the statistical assumptions underlying this usage. In an attempt to illustrate the complexities of this statistical technique, three sham studies using actual patient data are presented. These staged simulations have varying relationships between group test performance differences and levels of covariate discrepancy. The results were robust and consistent in their nature, and were held to support the wisdom of previous cautions by statisticians concerning the employment of ANCOVA to justify comparisons between incomparable groups. ANCOVA should not be used in neuropsychological research to equate groups unequal on variables such as age and education or to exert statistical control whose objective is to eliminate consideration of the covariate as an explanation for results. Finally, the report advocates by example the use of simulation to further our understanding of neuropsychological variables.

Adult↗

Slow-growing Rhizobium japonicum comprises two highly divergent symbiotic types.

We examined the interrelationships of the genomes of 10 slow-growing strains of Rhizobium japonicum to provide a foundation for molecular genetic studies of these agriculturally important endosymbiotic bacteria of commercial soybeans. The degree of base substitution in and around known symbiotic genes (nif and presumptive nod), constitutively expressed genes (glnA and recA), and two other cloned sequences was estimated from restriction site variation by using cloned DNAs as hybridization probes to genomic Southern blots. Two highly divergent patterns of conservation of nifDH genes and nod-homologous sequences were found. On this basis, we classified the strains as the symbiotic genotypes sTI or sTII. Existing maps of the nif genes of R. japonicum apply only to strains of the sTI genotype. This division was further characterized by four other probes which also distinguished two sublines within sTI. Phenograms were constructed depicting interrelationships according to DNA sequence divergence. sTI and sTII are two highly divergent evolutionary lines consistent with the status of individual species. Neither is related to fast-growing Rhizobium strains (PRC strains) nodulating soybeans.

Chromosome Mapping↗

The evolution of a plant globin gene family.

We have analyzed the sequences of soybean leghemoglobin genes as an initial step toward understanding their mode of evolution. Alignment of the sequences of plant globin genes with those of animals reveals that based on the proportion of nucleotide substitutions that have occurred at the first, second, and third codon positions, the time of divergence of plant and animal globin gene families appears to be extremely remote (between 900 million and 1.4 billion years ago, if one assumes constancy of evolutionary rate in both the plant and animal lineages) and in addition to the normal regulatory sequences on the 5' end, an approximately 30-base-pair sequence, specific to globin genes, that surrounds the cap site is conserved between the plant and animal globin genes. Comparison of the leghemoglobin sequences with one another shows that the relative amount of sequence divergence in various coding and noncoding regions is roughly similar to that found for animal globin genes and as in animal globin genes, the positions of insertions and deletions in the intervening sequences often coincide with the locations of direct repeats. Thus, the mode of evolution of the plant globin genes appears to resemble, in many ways, that of their animal counterparts. We contrast the overall intergenic organization of the plant globin genes with that of animal genes, and discuss the possibility of the concerted evolution of the leghemoglobin genes.

Animals↗

Can neuropsychological tests detect the sites of cerebrovascular stenoses and occlusions?

Conventional neuropsychological data may not mirror the site of cerebrovascular stenosis in a patient with mild to moderate symptoms. In this study, three groups of cerebral revascularization candidates failed to differ on an extended Halstead-Reitan battery: patients with symptoms and angiographic results referable to the carotid arterial system, subjects with clinical signs and neuroradiological findings referable to the vertebrobasilar arterial system, and patients with symptoms of cerebral ischemia but negative angiograms. Consistent with previous work, these patients generally were mildly impaired on neuropsychological tests, even though many were not symptomatic at the time of assessment. Neuropsychological tests seem to be sensitive to the presence and severity of cerebral ischemia, but in isolation may not be sufficient to determine the loci of vascular stenoses. The incongruence between angiographic and neuropsychological findings underlines the importance of distinguishing among different levels and types of measurements in defining subgroups of cerebral revascularization candidates. Collaboration among neuroscientists will further our understanding of the interrelationships among neurodiagnostic tests.

Carotid Artery Diseases↗

Neurobehavioral deficit and computed tomographic abnormalities in three samples of schizophrenic patients.

Recent reports have suggested that schizophrenics show brain abnormalities as reflected on computed tomography (CT Scan). We present data on three groups of schizophrenic patients which replicate our original report of this phenomenon with associated neuropsychological test results. The over-all evidence for an increased rate of neuropsychological abnormalities and related CT structural anomalies in groups of schizophrenics in several studies is robust. The biological dynamics, cause-effect relationships, and clinical applicability of this phenomenon in the individual case, however, are far from clear. Diagnostic agreements between behavioral and structural abnormalities should simultaneously sound a note of clinical caution and stimulate further study.

Brain Damage, Chronic↗

Rat mitochondrial DNA polymorphism: sequence analysis of a hypervariable site for insertions/deletions.

Three closely related variants of rat (Rattus norvegicus) mtDNA have been shown to differ in the number of T residues found in a run of Ts (light strand) which spans the junction between the tRNACys and tRNATyr genes. The number of Ts in the repeat varies from 6 to 8 in these DNAs. Another, less closely related, R. norvegicus variant has a run of 5 Ts at this site and in the related species, Rattus rattus, a run of 4 Ts is found. In R. norvegicus mtDNA runs of 5 As and 5 Gs are found just to the 3' side of the variable T repeat, and it is suggested that the three runs of repeated nucleotides may stabilize heteroduplexes which result from strand slippage and which give rise to the insertions and/or deletions. Among 17 mtDNA clones derived from an individual with the 8T repeat, one clone was found which possessed a 9T repeat. This variant may represent an additional DNA type originally present within the individual.

Animals↗

Chromosomal arrangement of leghemoglobin genes in soybean.

A cluster of four different leghemoglobin (Lb) genes was isolated from AluI-HaeIII and EcoRI genomic libraries of soybean in a set of overlapping clones which together include 45 kilobases (kb) of contiguous DNA. These four genes, including a pseudogene, are present in the same orientation and are arranged in the order: 5'-Lba-Lbc1-Lb psi-Lbc3-3'. The intergenic regions average 2.5 kb. In addition to this main Lb locus, there are other Lb genes which do not appear to be contiguous to this locus. A sequence probably common to the 3' region of Lb loci was found flanking the Lbc3 gene. The 3' flanking region of the main Lb locus also contains a sequence that appears to be expressed more abundantly in root tissue. Another sequence which is primarily expressed in root and leaf is found 5' to two Lb loci. Overall, the main leghemoglobin locus is similar in structure to the mammalian globin gene loci.

Base Sequence↗

Effect of lectins and sugars on primary sperm attachment in the horseshoe crab, Limulus polyphemus L.

The apical regions of motile Limulus spermatozoa readily adhere to the outer layer of the egg envelope. Shortly after this adherence or primary attachment, the sperm acrosome reaction occurs, resulting in a stronger adhesion (secondary attachment). A sperm attachment assay that quantified the number of spermatozoa attaching to egg sections was utilized to identify components involved in primary attachment. The number of spermatozoa attached was examined after treatment of either egg sections or spermatozoa with various compounds. Egg sections treated with asparagus pea lectin (250 micrograms/ml) bound significantly fewer spermatozoa as compared to those exposed to wheat germ agglutinin, concanavalin A, and garden pea lectin. Furthermore, sperm attachment was also greatly reduced when egg sections were first incubated with the glycosidase, alpha-L-fucosidase (less than or equal to 5% of controls). Treatment of spermatozoa with alpha-L-fucose, fucoidan, or p-aminophenyl fucoside also reduced sperm attachment when compared to Millipore-filtered artificial seawater controls. Egg sections were treated with fluorescein-conjugated lectins to confirm that the lectins actually bound to portions of the egg envelope and that various sugars are present in the egg envelope. Evidence suggests that the methylpentose, alpha-L-fucose, plays an important role in primary sperm attachment in Limulus.

Animals↗

Novel features of animal mtDNA evolution as shown by sequences of two rat cytochrome oxidase subunit II genes.

The sequence of the region of the mitochondrial genome that encodes cytochrome oxidase subunit II (COII) has been determined for each of two closely related rat species, Rattus norvegicus and R. rattus. Comparison of the two sequences shows that 94.4% of the nucleotide substitutions are silent. The occurrence of this high proportion of silent substitutions leads us to propose that the rapid evolution of mtDNA relative to nuclear DNA is due only to silent changes and that amino acid-altering substitutions accumulate in nuclear and mtDNA at comparable rates. Other novel features of the nucleotide substitution pattern in the rat COII gene are a high transition/transversion ratio (8.0:1) and a strong bias toward C in equilibrium T transitions in the light strand. Comparison of the R. norvegicus COII sequence with the bovine and human sequences show that there may be selective constraints on some silent positions within the gene and that its rate of evolution may be different in different mammalian lineages.

Animals↗

Preoperative neurobehavioral impairment in cerebral revascularization candidates.

We examined level of performance in 40 candidates for cerebral revascularization and found substantial variability on general indices of neuropsychological performance. Patients differed by duration of their longest ischemic attack, by surgery type, but not by time since onset of their first ischemic attack. Duration of the longest ischemic attack correlated highly with neuropsychological summary scores. In contrast, symptom duration was unrelated to level of performance in 18 additional patients with histories consistent with cerebrovascular ischemia who did not undergo surgery subsequently. We concluded that initial status on neuropsychological tests is an important variable to consider in postoperative followup of patients undergoing cerebral revascularization. More detailed clinical history and finer grained analysis of neuropsychological data promised to reveal additional relationships among clinical symptoms, neurobehavioral data, and underlying neuropathology.

Brain Ischemia↗

Memory performance of chemical workers exposed to polybrominated biphenyls.

Twenty-five chemical workers who manufactured polybrominated biphenyls (PBB's) were given objective tests of learning and memory. Although this group had high concentrations of PBB's in adipose tissue, mean scores on all memory tests were normal. The PBB concentration was not correlated with memory performance; the most contaminated workers showed no evidence of memory dysfunction.

Adult↗