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Responses of motor units during the hind limb flexion withdrawal reflex evoked by noxious skin heating: phasic and prolonged suppression by midbrain stimulation and comparison with simultaneously recorded dorsal horn units.

In rats anesthetized with sodium pentobarbital, we quantitatively analyzed descending modulation from the midbrain of a nociceptive flexion withdrawal reflex and responses of associated spinal neurons. We monitored the isometric force of hind limb withdrawal elicited by noxious heat stimuli (42-54 degrees C, 10 sec) on the hind paw. In one series of experiments, single-fiber EMG electrodes recorded responses of single muscle fibers (i.e., motor units) in biceps femoris during the hind limb withdrawal, without and during electrical stimulation in the midbrain periaqueductal gray (PAG) or lateral midbrain reticular formation (LRF). In a second series, responses of single lumbar dorsal horn neurons were also recorded simultaneously. Withdrawal force and associated motor unit responses were suppressed for prolonged periods (4 to greater than 60 min) following the initial episode of PAG or LRF stimulation in 40% of the rats, while they were suppressed phasically (i.e., only during brain stimulation) in the remainder. Motor unit responses increased in a graded fashion with increasing skin stimulus temperature from threshold (45 degrees C) to 54 degrees C. During PAG stimulation, the slope of the rate coding function was reduced with no change in threshold temperature. During LRF stimulation the rate coding function was shifted toward higher temperatures with increased threshold (47 degrees C). In 14 experiments 43 paired recordings were made from a dorsal horn and a motor unit during hind limb withdrawals. Mean latency to onset and peak of the heat-evoked response was shorter for dorsal horn compared to motor units. In 6/14 rats withdrawal force and motor unit responses were significantly suppressed for more than 8 min following mechanical placement of the stimulating electrodes and/or the initial episode of midbrain stimulation, while the simultaneously recorded dorsal horn unit responses remained constant. Following supplemental administration of pentobarbital (10-30 mg/kg i.v.), withdrawals and motor unit responses to heat were suppressed while dorsal horn unit responses were unchanged or enhanced. Also, in 12/42 cases, withdrawals and motor unit responses decremented markedly during the initial 3 trials of heat, while simultaneously recorded dorsal horn unit responses remained stable. These results indicate that the withdrawal reflex and associated motor units can be markedly suppressed in the absence of concomitant changes in responsiveness of dorsal horn neurons, and are discussed in terms of the neurocircuitry of spinal flexor reflexes and their descending modulation.

Animals↗

Detecting natural selection on cis-regulatory DNA.

Changes in transcriptional regulation play an important role in the genetic basis for evolutionary change. Here I review a growing body of literature that seeks to determine the forces governing the non-coding regulatory sequences underlying these changes. I address the challenges present in studying natural selection without the familiar structure and regularity of protein-coding sequences, but show that most tests of neutrality that have been used for coding regions are applicable to non-coding regions, albeit with some caveats. While some experimental investment is necessary to identify heritable regulatory variation, the most basic inferences about selection require very little functional information. A growing body of research on cis-regulatory variation has uncovered all the forms of selection common to coding regions, in addition to novel forms of selection. An emerging pattern seems to be the ubiquity of local adaptation and balancing selection, possibly due to the greater freedom organisms have to fine-tune gene expression without changing protein function. It is clear from multiple single locus and whole genome studies of non-coding regulatory DNA that the effects of natural selection reach far beyond the start and stop codons.

Animals↗

Methodological issues in coding sleep states in immature infants.

Thirty-five healthy, premature infants, ranging from 30-39 weeks postconceptional age, were observed continuously for 6 to 24 hr. Behavioral state and electroencephalographic patterns were coded for each minute. Using these data, three questions regarding coding of states of sleep were addressed: What is the concordance between behavioral codes and specific EEG patterns? Does the concordance between behavioral codes and EEG patterns change with postconceptional age? What range of error can be expected when observation periods shorter than 24-hr are used to estimate the daily distribution of quiet sleep (QS) and active sleep (AS)? With behavioral codes as the standard, concordances of EEG patterns for QS and AS were 72.5 and 92.1% respectively. With EEG patterns as the standard, behavioral codes for QS and AS agreed 83.0 and 88.9%. Agreement between behavioral codes and EEG patterns for QS increased with age. Finally, variation in estimates of the daily distribution of QS and AS decreased dramatically as the length of observation increased from 3 to 24 hr.

Arousal↗

An on-line interactive interviewing program for epidemiological studies.

An on-line interactive verification and entry system (OLIVES) has been developed for conducting telephone interviews in large-scale epidemiological studies. Responses are automatically coded into a computer legible form suitable for analysis. Use of a question stack to control question flow allows on-line response modification, restart from any termination point, and minimal reprogramming in order to change question order. Numerical, coded, and text string data types are permitted. A multilevel hierarchical data structure reflects interview content. The advantages and disadvantages of the on-line approach are discussed. OLIVES is generalizable and is applicable to other on-line or off-line interview situations.

Child↗

Demonstration of loss of attenuation and extended field persistence of a live avian metapneumovirus vaccine.

A live A type avian metapneumovirus (AMPV) vaccine which had been shown to be highly protective and short lived in experimental conditions was found to persist for longer periods in the field and to be associated with disease. Previously other factors such as possible secondary pathogens and management considerations had made it impossible to conclude whether the observed disease was a result of an increase in the vaccine virulence. In this study, an AMPV was isolated from poults on a farm which had been vaccinated with the same live A type vaccine. Full sequencing of the isolate, the vaccine and the vaccine progenitor confirmed its vaccine origin and further showed that generation of the vaccine had only involved nine substitutions of which three coded for amino acid changes. The isolated virus was inoculated into 1-day-old turkey poults in disease secure isolators and shown to cause disease with a severity similar to that caused by virulent field virus. Only two coding mutations were associated with this reversion to virulence.

Animals↗

Trends in hospital activity, morbidity and case fatality related to atrial fibrillation in Scotland, 1986--1996.

AIMS: Atrial fibrillation is a common and important cause of cardiovascular morbidity and mortality that may become more prevalent due to an ageing population and more prolonged exposure to predisposing cardiovascular disease states. This study examines recent trends in hospitalizations related to atrial fibrillation in Scotland. METHODS AND RESULTS: Scotland (population 5.1 million) has a well described system for recording hospitalization data. All hospital discharges (and death) can be linked for each individual patient. We examined the period 1986--1996, during which time a total of 103,085 hospitalizations with a principal or secondary diagnosis of atrial fibrillation were recorded. The number of hospitalizations with a principal diagnosis of atrial fibrillation increased threefold from 1869 in 1986 to 5757 in 1996; the number with a secondary diagnosis rose from 3577 to 11,522. Similar increases were seen in the number of patients hospitalized, in those having a 'first-ever' hospitalization and in population hospitalization rates overall. The average age of patients rose, in men from 63.8 (SD 13.2) to 65.0 (13.2) years and in women from 72.2 (12.2) to 73.2 (11.4) years. The proportion of those aged >75 years rose from 33% to 35% in men and from 56% to 60% in women. Average length of stay and case fatality fell during this period, but, because of the overall increase in hospitalizations, atrial fibrillation contributed to a growing proportion of cardiovascular-related bed-days utilized (from 18% to 37% with atrial fibrillation coded in any diagnostic position). CONCLUSION: The number of hospitalizations for atrial fibrillation has increased dramatically (two- to threefold) in recent years. These findings may be due to a real increase in atrial fibrillation prevalence, changing medical practice (e.g. coding or admission thresholds) or both. Consequently, the public health burden of atrial fibrillation is enormous. Moreover, the observed increase in atrial fibrillation-related hospital activity shows no sign of abating.

Age Distribution↗

Editing of transfer RNAs in Acanthamoeba castellanii mitochondria.

With the discovery of RNA editing, a process whereby the primary sequence of RNA is altered after transcription, traditional concepts of genetic information transfer had to be revised. The known RNA editing systems act mainly on messenger RNAs, introducing sequence changes that alter their coding properties. An editing system that acts on transfer RNAs is described here. In the mitochondria of Acanthamoeba castellanii, an amoeboid protozoan, certain transfer RNAs differ in sequence from the genes that encode them. The changes consist of single-nucleotide conversions (U to A, U to G, and A to G) that appear to arise posttranscriptionally, are localized in the acceptor stem, and have the effect of correcting mismatched base pairs. Editing thus restores the base pairing expected of a normal transfer RNA in this region.

Acanthamoeba↗

Accounting for evolutionary rate variation among sequence sites consistently changes universal phylogenies deduced from rRNA and protein-coding genes.

Phylogenetic analyses of gene and protein sequences have led to two major competing views of the universal phylogeny, the evolutionary tree relating the three kinds of living organisms, Bacteria, Archaea, and Eukarya. In the first scheme, called "the archaebacterial tree, " organisms of the same type are clustered together. In the second scenario, called "the eocyte tree," the archaeal phylum of Crenarchaeota is more closely related to eukaryotes than are other Archaea. A major property of the evolution of functional ribosomal and protein-encoding genes is that the rate of nucleotide and amino acid substitution varies across sequence sites. Here, using distance-based and maximum-likelihood methods, we show that universal phylogenies of ribosomal RNAs and RNA polymerases built by ignoring this variation are biased toward the archaebacterial tree because of attraction between long branches. In contrast, taking among-site rate variability into account gives support for the eocyte tree.

Animals↗

The importance of minor abnormalities in the resting electrocardiogram.

Minor abnormalities of the ST segment and T wave are seen in about 1% of the resting electrocardiograms of the 'normal' population. Such changes are found more commonly in people of African descent and with increasing age. In young people they are often not present when the recording is repeated. In epidemiological studies minor ST segment and T wave (ST-T) electrocardiographic changes have been found to be associated with an increased risk of death. However, a sizeable minority of subjects with such abnormalities have symptoms suggesting ischaemic heart disease, and the presence of such symptoms is much more important as a risk predictor than are the electrocardiographic changes themselves. It is important that 'non-specific' ST-T wave changes should be accurately documented, perhaps using the Minnesota code. Aircrew found to have such changes should have an effort test, but if this is normal and if they are asymptomatic their licences need not be restricted.

Adult↗

Evaluation of quantitative parametric analysis for characterization of breast lesions in contrast-enhanced MR mammography.

Our aim was to evaluate quantitative parametric analysis for characterization of breast lesions in contrast-enhanced magnetic resonance (MR) mammography. In 62 patients, contrast-enhanced MR mammography revealed 75 suspicious lesions, of which 18 were benign and 57 were malignant. The quantitative parametric analysis delineates signal intensity changes of contrast-enhanced lesions on a pixel-by-pixel basis. The initial rate of enhancement is coded by color intensity: a slow rate is coded to dark; a fast rate, bright. The postinitial enhancement change is coded by color hue: blue for increasing signal intensity, green for plateau, and red for decrease in signal intensity. Malignant lesions showed a significantly higher number of bright-red (P = 0.004) and medium-red (P < 0.001) pixels than benign lesions. Benign lesions showed significantly more blue pixels than did malignant lesions (P = 0.010). Of the 75 lesions, 72 (96%) showed heterogeneous distribution of pixel color hue. Quantitative parametric analysis of contrast kinetics in lesions can replace the subjective manual region of interest (ROI) method and makes a step toward standardization of MR mammography. It allows quantitative evaluation of different contrast kinetics parameters in contrast-enhanced breast lesions.

Adult↗

Temperature-sensitive mutations in the gene encoding the small subunit of the vaccinia virus early transcription factor impair promoter binding, transcription activation, and packaging of multiple virion components.

The vaccinia virus D6R open reading frame encodes the small subunit of the heterodimeric vaccinia virus early transcription factor (VETF) that activates transcription of early genes in vitro. VETF binds early gene promoters and has a DNA-dependent ATPase activity that is essential for activation of transcription. To examine the relationship between the structure and function of VETF, we have localized the mutations in two temperature-sensitive viruses whose lesions previously were mapped to the D6R gene. For both mutants, a single G-to-A nucleotide change that would alter protein coding potential was identified. In mutant E93, the codon for alanine 25 was changed to that of threonine, and in mutant S4 the codon for valine 278 was replaced with that for methionine. The molecular phenotype of each mutant was assessed by expressing mutant transcription factors in HeLa cells by using a vaccinia virus-T7 system and characterizing the proteins' activities in vitro. The A25T mutant activated transcription to a lesser extent than wild-type VETF, and the V278M mutant had no demonstrable transcription factor activity. Both mutant proteins were shown to be defective for promoter binding, accounting for their impairment in transcription activation. The functional defects for both mutants were observed at permissive as well as nonpermissive temperatures. The mutant proteins retained ATPase activity but required higher DNA concentrations to activate the ATPase. These results indicate that the small subunit of VETF is essential for its promoter binding activity and likely contacts the promoter DNA. Immunoblotting experiments showed that the virion particles from the two mutant viruses contained about half the VETF of wild-type virus, suggesting that promoter binding may contribute to packaging of VETF into the virion particle. RNA polymerase, mRNA capping enzyme, and nucleoside triphosphate phosphohydrolase I were found at similarly reduced levels in the virion, indicating that packaging of some virion core enzymes may be interdependent.

Adenosine Triphosphatases↗

Molecular basis for plasma alpha(1,3)-fucosyltransferase gene deficiency (FUT6).

While most humans express an alpha(1,3)-fucosyltransferase in plasma, 9% of individuals on the isle of Java (Indonesia) do not express this enzyme. Ninety-five percent of these plasma alpha(1,3)-fucosyltransferase-deficient individuals have Lewis negative phenotype on red cells, suggesting strong linkage disequilibrium between these two traits. To define the molecular basis for this plasma deficiency and to determine which of two candidate human alpha(1,3)-fucosyltransferase genes encode this enzyme (FUT5 and FUT6), we cloned and analyzed alleles at these two loci from an Indonesian individual deficient in plasma alpha(1,3)-fucosyltransferase activity. Single base pair changes were identified in the coding region of each gene, relative to previously published wild type alleles. These changes in turn yield three codon changes in FUT5 and three in FUT6. The codon changes in the FUT5 gene do not yield detectable diminutions in alpha(1,3)-fucosyltransferase activity when tested by expression in transfected COS-1 cells, and none of the FUT5 alleles co-segregate with plasma alpha(1,3)-fucosyltransferase deficiency in Indonesian pedigrees. By contrast, two of the codon changes in the FUT6 alleles inactivate this gene when tested by expression in transfected COS-1 cells. One of these inactivating changes is a missense mutation (Glu-247-->Lys) within the enzyme's catalytic domain. The other inactivating mutation represents a nonsense mutation (Tyr-315-->stop) that truncates the COOH terminus of the enzyme by 45 amino acids. The Glu-247-->Lys missense mutation is present in double dose in the nine plasma alpha(1,3)-fucosyltransferase-deficient individuals tested, whereas the nonsense mutation at tyrosine 315 is present in double dose in just one of these persons. These results demonstrate that the alpha(1,3)-fucosyltransferase activity in human plasma is encoded by the FUT6 gene and that the missense mutation within codon 247 of this gene is responsible for deficiency of this activity in these Indonesian families.

Alleles↗

[Value of color-coded Doppler sonography in the differential diagnosis of nodular thyroid gland changes].

AIM: The incidence of functional autonomous adenomas, detected in every second nodular goiter by scintigraphic methods is very high in an area of iodine deficiency. The color-coded Doppler sonography (CCDS) as a diagnostic tool in differentiating thyroid nodules is discussed controversially. METHODS: In this prospective study we investigated the value of CCDS in 200 patients with nodular thyroid alterations compared with 99m-Technetium (Tc) scintigraphy. RESULTS: Focal maximas of Tc-uptake were detected in 22.5% of all patients, and 44.5% of the thyroid nodules showed increased vascularity. There was no correlation between nodular vascularity and thyroid 99m-Tc uptake (TcTU). In contrast to this we could demonstrate a significant relation between vascularity and the diameter of the nodule (p < 0.0001). The results are discussed in the context of method specific limitations of ultrasound examinations. CONCLUSION: Our results confirm that CCDS has no great importance in the differentiation of thyroid nodules. Scintigraphy remains the diagnostic method of choice to assess the topographic thyroid function.

Adenoma↗

An investigation of the increase in preschool-age asthma in Manitoba, Canada.

Asthma has long been a major cause of illness and disability among young Canadians. From 1970-71 to 1987-88, hospital admissions for asthma increased significantly among Canadian children under the age of fourteen. Many hypotheses may explain this increase in asthma prevalence. There could be a true increase in the number of people developing symptoms of the disease or increased asthma rates could be an artifact due to changes in detection, diagnosis, treatment, or coding. This study reviews hypotheses put forward to explain the increase in asthma prevalence, and tests some of them in Manitoba for children aged 0-4. Physician claims data and hospital separation data were merged to create unique person oriented medical records. These records were used to estimate the number of children seeking medical services for asthma during a five-year period (1984-85 to 1988-89) and the change in rates over this time period. From 1984-85 to 1988-89, both prevalence and incidence rates for children less than five years of age increased. Prevalence rates showed strong seasonal peaks in the spring and the fall. There is no indication that asthma increased in severity. The hospitalization rate (the number visiting a hospital for asthma divided by the total number seeking medical care for asthma), the average number of hospital admissions per year, and the average number of days spent in a hospital per year did not increase. Levels of ozone (O3) and nitrogen dioxide (NO2) in downtown Winnipeg increased over the study period and asthma prevalence increased twice as fast in Winnipeg as in the rest of the province. For Manitoba, the increase in preschool-aged asthma does not appear to be due to increased use of medical services, a change in ICD coding, an increase in the severity of the cases, or a decrease in income levels. The increases appear to be at least partly due to changes in diagnostic practices. The relationship between asthma and air pollution needs more detailed study as pollution is likely to be an important factor, particularly during the spring. Other areas for further investigation are changes in allergy and virus precursors, maternal smoking, and increased levels of pollens, molds and dust mites.

Abstracting and Indexing↗

Symmetry preservation in the evolution of the genetic code.

The standard genetic code is found to exhibit an exact symmetry under a finite group of order 4 known in mathematics as the Klein group. The same symmetry is also present in almost all non-standard codes, mitochondrial as well as nuclear. Analysis of the phylogenetic tree for the evolution of the mitochondrial codes reveals that all changes along the main line of evolution preserve this symmetry, with a tendency towards symmetry enhancement. In the side branches of the evolutionary tree, the majority of changes also respect the symmetry. The few exceptional cases where it is broken correspond to reassignments that appear to be unstable or incomplete. Since the Klein group emerges naturally from the symplectic model for the prebiotic evolution that has led to the standard code, we interpret these results as lending support to the hypothesis that this symmetry has been selected during the evolution of the genetic code, not only before but also after establishment of the standard code.

Animals↗

beta 0 thalassemia, a nonsense mutation in man.

We determined the complete nucleotide sequence of the 5' noncoding region and the first 74 amino acids of the nonfunctional beta-globin mRNA in a patient with homozygous beta 0 thalassemia. We identified the molecular defect as a single nucleotide substitution in the coding region of the mRNA. At the position corresponding to amino acid 17, replacement of an adenine by a uracil changes the triplet AAG, which codes for lysine in the normal beta chain, to an amber termination codon, UAG. This type of beta 0 thalassemia represents an example of a nonsense mutation in man.

Base Sequence↗

Differential regulation of the messenger RNA for three major senescence marker proteins in male rat liver.

Changes in the mRNAs coding for specific hepatic proteins in male rats during aging were examined by in vitro translation of the liver mRNA in the rabbit reticulocyte lysate. Characterization of the [35S]methionine-labeled translation products by sodium dodecyl sulfate-polyacrylamide slab gel electrophoresis followed by autoradiography showed major age-dependent changes in the hepatic concentrations of three mRNA species. The translation products of these three mRNAs were found to have Mr = 28,500, 26,300, and 19,500 and are called senescence marker proteins (SMP) 1, 2, 3, respectively. On the basis of its immunochemical reactivity, SMP-3 (Mr = 19,500) is identified as alpha 2u-globulin while the mRNAs for SMP-1 (Mr = 28,500) and SMP-2 (Mr = 26,300) code for two yet uncharacterized proteins. The liver of the prepubertal male rats was found to contain the mRNA for SMP-2 and showed almost complete absence of the mRNAs for SMP-1 and SMP-3. However, the mRNAs for both SMP-1 and SMP-3 were present in the postpubertal young adults while the mRNA for SMP-2 was absent. Finally, when the animals reached senescence, the mRNAs for SMP-1 and SMP-3 disappeared from the liver with the reappearance of the SMP-2 mRNA. Age-dependent regulation of the mRNAs for these three senescence marker proteins can serve as an important model for the study of differential gene expression during aging.

Aging↗

Mitochondrial genomes of two demosponges provide insights into an early stage of animal evolution.

Mitochondrial DNA (mtDNA) of multicellular animals (Metazoa) is typically a small ( approximately 16 kbp), circular-mapping molecule that encodes 37 tightly packed genes. The structures of mtDNA-encoded transfer RNAs (tRNAs) and ribosomal RNAs (rRNAs) are usually highly unorthodox, and proteins are translated with multiple deviations from the standard genetic code. In contrast, mtDNA of the choanoflagellate Monosiga brevicollis, the closest unicellular relative of animals, is four times larger, contains 1.5 times as many genes, and lacks mentioned peculiarities of animal mtDNA. To investigate the evolutionary transition that led to the specific organization of metazoan mtDNA, we determined complete mitochondrial sequences from the demosponges Geodia neptuni and Tethya actinia, two representatives of the most basal animal phylum, the Porifera. We found that poriferan mtDNAs resemble those of other animals in their compact organization, lack of introns, and a well-conserved animal-like gene order. Yet, they contain several extra genes, encode bacterial-like rRNAs and tRNAs, and use a minimally derived genetic code. Our findings suggest that the evolution of the typical metazoan mtDNA has been a multistep process in which the compact genome organization and the reduced gene content were established prior to the reduction of tRNA and rRNA structures and the introduction of multiple changes of the translation code.

Amino Acid Sequence↗