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PubMed1993

Biological zero in laser Doppler measurements in normal, ischaemic and inflamed human skin.

The Biological Zero (BZ) signal in laser Doppler flowmetry was recorded during temporary occlusion of the arterial circulation to the limb by a blood pressure cuff inflated suprasystolically for 2 min. The non-thermoregulatory skin at the ankle was compared with the thermoregulatory skin over the toe pad in patients with lower limb arterial obstructive disease, elderly controls without arterial disease and young healthy adults under cool (15-17 degrees C) and warm (25-27 degrees C) ambient conditions. The absolute BZ values did not differ significantly within the three groups at the two room temperatures. However the resting laser Doppler flowmetry signal (LDflux) was consistently higher under warm than cold room temperatures: these differences were much greater at the toe than at the ankle. Accordingly, the relative size of BZ, expressed as a percentage of the resting LDflux, was least over the toe in a warm room (5-10%) and greatest over the malleolus (59-83%) in the cold room. The changes during the evolution of a positive tuberculin skin reaction were followed as a model of chronic inflammation in man. The resting LDflux rose 10 fold (from 0.3 to 3.4V) while the change in the BZ was minor (0.16-0.2V). Thus the rise in LDflux in a fully developed tuberculin reaction was approximately 30X greater than that for the corresponding BZ. Concurrent measurements of the alternative output from the laser Doppler flowmeter (CMBC) showed in a similar pattern. Since the BZ signal can be a substantial proportion of the resting LDflux measurement, we support the recommendation that the BZ should be measured wherever possible and practicable, for subtraction from the observed LDflux to assess the true flow: 'raw' LDflux measurements should be interpreted with extreme caution, particularly at relatively low resting LDflux levels.

Adolescent↗
PubMed1989

Unusual features of cereal seed protein structure and evolution.

The alcohol-soluble (prolamin) storage proteins of barley, wheat and rye vary in their structures, but all have two features in common: the presence of distinct structural domains differing in amino acid compositions, and of repeats within one of these domains. Detailed comparisons of amino acid sequences show that all appear to have evolved from a single ancestral gene consisting of three short related regions (called A, B and C). Regions related to A, B and C are also present in the minor prolamins of maize and in three other groups of seed proteins: inhibitors of alpha-amylase and/or trypsin from cereals. 25 storage globulins from several dicotyledonous species and a 2S albumin from sunflower. It is suggested that these proteins together constitute a protein superfamily with limited sequence homology.

Amino Acid Sequence↗
PubMed1988

Structural organization and evolution of the plastid genome of Vaucheria sessilis (Xanthophyceae).

The plastid DNAs of 18 Vaucheria sessilis strains from various habitats in western Europe were digested with the restriction endonucleases Eco RI, Sal I, Bam HI and Pvu II. Their restriction patterns showed variable fragment divergencies. Two main groups of plastid genomes were recognized, which were substantiated by morphological features. The differences among the restriction patterns could be attributed to the loss or appearance of restriction sites and to minor size variations caused by deletions/insertions. The Sal I and Bam HI restriction sites which together discriminate six different plastid genomes were mapped on the circular molecule of 124 kilobase paris (kbp). The plastid genomes of several Vaucheria sessilis strains were shown to exist in two inversion isomers caused by intramolecular recombination within the inverted repeat segments.

Biological Evolution↗
PubMed1989

The Nature of Light-Induced Inhibition of Photosystem II in Pumpkin (Cucurbita pepo L.) Leaves Depends on Temperature.

Attached leaves of pumpkin (Cucurbita pepo L.) were treated in high or moderate light at room temperature or a 1 degrees C. The symptoms of photoinhibition appearing during light treatments at room temperature could be attributed to a decrease in the primary activity of PSII. However, when the light treatment was given at 1 degrees C, the quantum yield of photosynthetic oxygen evolution decreased much more than would be expected from the decrease in the ratio of variable to maximum fluorescence at 77 degrees K. Also, light treatment at 1 degrees C lowered the chloroplast wholechain electron transfer capacity much more than it affected PSII electron transport (H(2)O to paraphenylbenzoquinone). Light treatments at both room temperature and 1 degrees C led to an increase in B(max), which indicates an increase in the proportion of PSII(beta) centers. PSI was not affected by the light treatments, and the treatments in the dark at 1 degrees C caused only minor changes in the measured properties of the leaves. We conclude that high light always inhibits the primary activity of PSII, but at low temperature there is greater inhibition of electron transfer from primary electron accepting plastoquinone of PSII to the plastoquinone pool, which leads to a drastic decrease in the quantum yield of oxygen evolution in the chilling-sensitive pumpkin.

Journal Article↗
PubMed2004

Evolutionary implication of human endogenous retrovirus HERV-H family.

The human endogenous retrovirus (HERV-H) family is most abundant and widely distributed in the human genome, with about 100-1,000 full-length or deleted elements and a similar number of solitary, long-terminal repeats. The HERV-H env ORF has been characterized in humans and in the course of primate evolution, indicating the increased possibility of biological roles in humans. Using the polymerase chain reaction approach with a human monochromosomal DNA panel, 70 env fragments belonging to the HERV-H family from chromosomes 1, 2, 3, 4, 5, 6, 7, 9, 10, 11, 12, 14, 15, 16, 17, 18, 19, 20, X, and Y were identified and analyzed. They showed 82-99% sequence similarity to that of HERV-H (accession no. AF108843). We also identified other HERV-H env fragments in the DDBJ/EMBL/GenBank databases. The total of 120 fragments was evolutionarily analyzed. Phylogenetic analysis suggests that the HERV-H env family is divided into one major and two minor groups. The HERV-H members have been actively proliferated and evolved by intra-chromosomal spread during hominid radiation.

Chromosomes↗
PubMed1997

A cottontail rabbit papillomavirus strain (CRPVb) with strikingly divergent E6 and E7 oncoproteins: An insight in the evolution of papillomaviruses.

We previously observed that warts induced by an isolate of cottontail rabbit papillomavirus (CRPV) showed incomplete instead of systemic regression in some domestic rabbits. We report that the viral isolate contained, as a major component, a CRPV strain (CRPVb) showing an unexpectedly high divergence in the E6 and E7 open reading frames (ORFs), compared to the prototype CRPVa present in the isolate as a minor component. The E6 and E7 oncoproteins of CRPVa and -b disclosed only 87.5% identical amino acids and differed in size by three and two amino acids, respectively. This divergence involved (i) a great number (4.4%) of nucleotide substitutions and a high rate (83.3%) of nonsynonymous mutations; (ii) mutations changing the E6 and E7 stop codons; and (iii) in-frame sequence insertions in the E6 ORF (18 nucleotides) and downstream of the mutated E7 stop codon (6 nucleotides), both likely to result from a duplication of adjacent sequences. These extensive differences could account for distinct biological and antigenic properties. Strikingly, only four (0.8%) amino acids of the L1 major capsid protein were variable. Thus, it seems likely that sequence duplications and mutations affecting stop codons exert a strong selection pressure on the fixation of nonsynonymous mutations and that phylogenetic calculations based only on point mutations may misevaluate the time scale of the evolution of papillomaviruses.

Amino Acid Sequence↗
PubMed1990

Defective interfering particles: effects in modulating virus growth and persistence.

Defective interfering virus particles (DIP) frequently play an important part in viral persistence in vitro, and may in some instances modify a virus infection in vivo, causing attenuation or persistence of the infection. To explain certain aspects of the growth of these mutants in vitro, other factors have been invoked such as interferon, mutations in the wild-type virus or the infected cells, or other substances released by infected cells that attenuate the infection. We present here a simple model of the growth of DIP in vitro which shows that (a) the observed population dynamics of DIP can readily be explained without invoking such extrinsic factors; (b) the initial multiplicity of infection of DIP is the principal determinant of the outcome of infection in both single- and repeated-passage cultures; and (c) in a long-term culture in vitro, the criterion used to decide the time of virus passage directly determines how long the standard virus, DIP, and cells survive. This model may be used with minor modifications to predict the behavior in vitro of other mutant viruses with a dominantly interfering phenotype.

Biological Evolution↗
PubMed2005

Structures of the agouti signaling protein.

Expression of the agouti signaling protein (ASIP) during hair growth produces the red/yellow pigment pheomelanin. ASIP, and its neuropeptide homolog the agouti-related protein (AgRP) involved in energy balance, are novel, paracrine signaling molecules that act as inverse agonists at distinct subsets of melanocortin receptors. Ubiquitous ASIP expression in mice gives rise to a pleiotropic phenotype characterized by a uniform yellow coat color, obesity, overgrowth, and metabolic derangements similar to type II diabetes in humans. Here we report the synthesis and NMR structure of ASIP's active, cysteine-rich, C-terminal domain. ASIP adopts the inhibitor cystine knot fold and, along with AgRP, are the only known mammalian proteins in this structure class. Moreover, ASIP populates two distinct conformers resulting from a cis peptide bond at Pro102-Pro103 and a coexistence of cis/trans isomers of Ala104-Pro105. Pharmacologic studies of Pro-->Ala mutants demonstrate that the minor conformation with two cis peptide bonds is responsible for activity at all MCRs. The loop containing the heterogeneous Ala-Pro peptide bond is conserved in mammals, and suggests that ASIP is either trapped by evolution in this unusual configuration or possesses function outside of strict MCR antagonism.

Agouti Signaling Protein↗
PubMed1982

Epidural hematoma: report of seven cases with delayed evolution of symptoms.

Epidural hematomas occasionally have a prolonged clinical course with gradual evolution of the neurologic symptoms. Seven such cases are reviewed in this report. Although the clinical course is insidious, there are certain features which should signal the presence of a slowly expanding hematoma. After an apparently minor head injury, the patient who is usually in the younger age group, develops headache. This persists and is accompanied by other non-specific neurologic symptoms which may lead to a mistaken diagnosis of "post-concussion syndrome". Papilledema and focal neurologic deficits eventually appear. Definitive diagnosis is made by the CT scan, although contrast enhancement may be necessary to confirm the nature of the lesion. The treatment is craniotomy and evacuation of the hematoma before serious neurologic deterioration occurs.

Adolescent↗
PubMed2002

Consensus-derived structural determinants of the ankyrin repeat motif.

The ankyrin repeat is one of the most common, modular, protein-protein interaction motifs in nature. To understand the structural determinants of this family of proteins and extract the consensus information that defines the architecture of this motif, we have designed a series of idealized ankyrin repeat proteins containing one, two, three, or four repeats by using statistical analysis of approximately 4,000 ankyrin repeat sequences from the PFAM database. Biophysical and x-ray crystallographic studies of the three and four repeat constructs (3ANK and 4ANK) to 1.26 and 1.5 A resolution, respectively, demonstrate that these proteins are well-folded, monomeric, display high thermostability, and adopt a very regular, tightly packed ankyrin repeat fold. Mapping the degree of amino acid conservation at each position on the 4ANK structure shows that most nonconserved residues are clustered on the surface of the molecule that has been designated as the binding site in naturally occurring ankyrin repeat proteins. Thus, the consensus amino acid sequence contains all information required to define the ankyrin repeat fold. Our results suggest that statistical analysis and the consensus sequence approach can be used as an effective method to design proteins with complex topologies. These generic ankyrin repeat proteins can serve as prototypes for dissecting the rules of molecular recognition mediated by ankyrin repeats and for engineering proteins with novel biological functions.

Amino Acid Sequence↗
PubMed2002

Hill-Robertson interference is a minor determinant of variations in codon bias across Drosophila melanogaster and Caenorhabditis elegans genomes.

According to population genetics models, genomic regions with lower crossing-over rates are expected to experience less effective selection because of Hill-Robertson interference (HRi). The effect of genetic linkage is thought to be particularly important for a selection of weak intensity such as selection affecting codon usage. Consistent with this model, codon bias correlates positively with recombination rate in Drosophila melanogaster and Caenorhabditis elegans. However, in these species, the G+C content of both noncoding DNA and synonymous sites correlates positively with recombination, which suggests that mutation patterns and recombination are associated. To remove this effect of mutation patterns on codon bias, we used the synonymous sites of lowly expressed genes that are expected to be effectively neutral sites. We measured the differences between codon biases of highly expressed genes and their lowly expressed neighbors. In D. melanogaster we find that HRi weakly reduces selection on codon usage of genes located in regions of very low recombination; but these genes only comprise 4% of the total. In C. elegans we do not find any evidence for the effect of recombination on selection for codon bias. Computer simulations indicate that HRi poorly enhances codon bias if the local recombination rate is greater than the mutation rate. This prediction of the model is consistent with our data and with the current estimate of the mutation rate in D. melanogaster. The case of C. elegans, which is highly self-fertilizing, is discussed. Our results suggest that HRi is a minor determinant of variations in codon bias across the genome.

Animals↗
PubMed1998

Regional glucose utilization in infarcted and remote myocardium: its relation to coronary anatomy and perfusion.

We studied the relationship between coronary anatomy, perfusion and metabolism in myocardial segments exhibiting transient and persistent perfusion defects on stress/rest 99Tcm-MIBI single photon emission tomography in 35 patients (31 males, 4 females, mean age 56 +/- 7 years) with a previous myocardial infarction. Quantitative coronary angiography and assessment of myocardial perfusion reserve and glucose metabolism were performed within 1 week of one another. Perfusion was assessed by SPET after the intravenous injection of 740 MBq of 99Tcm-MIBI at rest and after exercise. Regional myocardial glucose metabolism was assessed by position emission tomography at rest (200 MBq of 18F-2-deoxyglucose, FDG) after an overnight fast with no glucose loading. All 35 patients exhibited persistent perfusion defects consistent with the clinically identified infarct site, and 27 (77%) also showed various degrees of within-infarct FDG uptake; 11 patients developed exercise-induced transient perfusion defects within, or in the vicinity of, 15 infarct segments and resting FDG uptake was present in 10 of these segments (67%). Five patients also showed exercise-induced transient perfusion defects in nine segments remote from the site of infarct: resting FDG uptake was present in six of these regions (67%). Finally, nine patients had increased glucose uptake in non-infarcted regions not showing transient perfusion defects upon exercise testing and perfused by coronary arteries with only minor irregularities. Our results confirm the presence of viable tissue in a large proportion of infarct sites. Moreover, FDG uptake can be seen in regions perfused by coronary arteries showing minor irregularities, not necessarily resulting in detectable transient perfusion defects on a MIBI stress scan. Since the clinical significance of such findings is not clear, further studies should be conducted to assess the long-term evolution of perfusion, function and metabolism in non-revascularized patients of those remote areas which are apparently normally perfused, but show abnormal fasting FDG uptake after myocardial infarction. Such studies may have important implications for the management of post-infarct patients, as the preservation of coronary vasodilator reserve and myocardial metabolism in remote myocardium may be seen as an additional goal in the treatment of such patients.

Blood Glucose↗
PubMed2002

Malignant liver tumors treated with MR imaging-guided laser-induced thermotherapy: experience with complications in 899 patients (2,520 lesions).

PURPOSE: To evaluate the complications from laser-induced thermotherapy (LITT) of malignant liver tumors and demonstrate that LITT is safe as an outpatient procedure. MATERIALS AND METHODS: During 8 years, 899 patients with malignant liver tumors were treated with magnetic resonance (MR) imaging-guided LITT. A total of 2,132 LITT procedures were performed to treat 2,520 lesions. To account for the technical evolution of LITT during this time and the change from performing the procedure on an inpatient basis to performing it on an outpatient basis, patients were assigned to four groups. Overall complication rates and major and minor complications in the inpatient versus outpatient groups were evaluated. Multidimensional contingency table analysis with the chi(2) test was performed. RESULTS: On the basis of a total of 2,132 LITT procedures performed, complications were divided into major and minor categories and detected at clinical or imaging studies. Major complications included three deaths (0.1%) within 30 days after LITT, pleural effusion requiring thoracentesis in 16 (0.8%) cases, hepatic abscess requiring drainage in 15 (0.7%) cases, bile duct injury in four (0.2%) cases, segmental infarction in three (0.1%) cases, and hemorrhage requiring transfusion in one (0.05%) case. Minor complications included postprocedural fever in 710 (33.3%), pleural effusion not requiring thoracentesis in 155 (7.3%), subcapsular hematoma in 69 (3.2%), subcutaneous hematoma in 24 (1.1%), pneumothorax in seven (0.3%), and hemorrhage in two (0.1%) cases. Outpatient management did not significantly affect pleural effusion (P =.96) or subcapsular hematoma (P =.33) rate. CONCLUSION: MR imaging-guided LITT with local anesthesia is safe and yields an acceptably low rate of major complications.

Ambulatory Care↗
PubMed2000

[Videolaparoscopic cholecystectomy. Experience and results in 1019 cases].

BACKGROUND: Laparoscopic cholecystectomy is now worldwide considered the elective therapy for biliary lithiasis. Morbidity, mortality and conversion rates reported in the literature are assessed between 1 and 6%, 0 and 0.9%, 3.6 and 7.2% respectively. Data on personal experience with 1019 attempted laparoscopic cholecystectomy are reported. METHODS: In the period between 1991 and 1997 1019 laparoscopic cholecystectomy were performed. Patients were 361 males and 658 females (ratio M:F 1:2), with an average age of 51 years (range 5-85). Indications were: 647 symptomatic cholelithiasis, 28 hydrops, 121 empyemas, 76 cholecystocholedocolithiasis and 13 alithiasic cholecystopathy. RESULTS: Conversion was necessary in 61 cases, with a conversion rate of 6%. Mean duration of surgery was 65 minutes (range 30-240) with a mean hospital stay of 2.1 days (range 1-10). No deaths occurred in our series, with a morbidity rate of 1.8% (18 cases, 7 major and 11 minor). Only 1 case of bile duct injury (0.1%) is reported. CONCLUSIONS: In consideration of low conversion rate, low early and late morbidity, absence of bile duct injury, advantages for the patient and the opportunity of evolution of this surgery, laparoscopic cholecystectomy can be considered the standard treatment for biliary lithiasis.

Adolescent↗
PubMed1985

Proliferation and migration of grafted hemopoietic cells during a graft-versus-host reaction induced by minor non-H-2 histocompatibility antigens in the mouse.

In our model of GVHR, irradiated (DBA/2 X B10.D2)F1 mice were given splenic and bone marrow cells from B10.D2 donor mice. At different set times after the graft, recipient mice were given a single injection of a radioactive precursor of DNA (125IUdR) and killed one hour later. The radioactivity in excised organs reflected the label incorporation by the proliferating cells. When mice were killed from 1 hour to 96 hours after the label injection the residual radioactivity in individual organs reflected the number of the residual living cells arising from cells which were in S-phase during the label pulse. This study allowed us to specify the dynamics of the cell proliferative activity and the behavior of these proliferating cells through the whole organism at any time of a GVH disease. A very interesting point is that the minor non-H-2 histocompatibility antigens (MiHA) responsible of the GVHR induced a very important and specific stimulation of the grafted cell proliferation in all the non-lymphoid organs with a spatial and timing evolution through the whole organism. A great part of the cells specifically stimulated to divide by the MiHA are very short-lived. The remaining living cells migrate out of the spleen and bone marrow and accumulate into the non-lymphoid organs after a latency period lasting 12-24 hours following the label uptake. This cell invasion was mainly into the liver, vesicular glands, kidneys, salivary glands and lungs.

Animals↗
PubMed1987

Long-term follow-up in untreated Plummer's disease (autonomous goiter).

Plummer's disease presents a spectrum of forms and its evolution is often slow. Longitudinal studies have been limited both in number of patients and duration of follow-up. In order to answer the question of whether euthyroid patients with no local symptoms should be treated prophylactically, 70 patients were reexamined 5.2-21.8 years (average, 8.9) after Plummer's disease had been diagnosed. Data are included on four others who developed symptoms that indicated treatment within five years. Gross (clinically relevant) changes were seen in 24 cases. Of these, 16 were progressive, including 12 where hyperthyroidism developed. Gross regressive changes included two cases with complete remission. Minor changes (28 cases) were about as often regressive as progressive. Most patients with a solitary autonomous nodule showed either gross changes or no change, whereas minor changes prevailed in multifocal autonomy. All eight male patients had a solitary nodule, 28 of 66 females had multifocal autonomy. Gross changes were seen in nine of 12 patients under 40 years at presentation, more often than in those over 49 years; seven of these changes were progressive. Six short case reports illustrate the great diversity in evolution. It is concluded that treatment seems wise in teenagers and in elderly patients with borderline (biochemical) hyperthyroidism but others generally may be left untreated, as long as they have few or no complaints.

Adenoma↗
PubMed1996

The evolution of fibrillar collagens: a sea-pen collagen shares common features with vertebrate type V collagen.

The extracellular matrix of marine primitive invertebrates (sponges, polyps and jellyfishes) contains collagen fibrils with narrow diameters. From various data, it has been hypothesized that these primitive collagens could represent ancestral forms of the vertebrate minor collagens, i.e., types V or XI. Recently we have isolated a primitive collagen from the soft tissues of the sea-pen Veretillum cynomorium. This report examines whether the sea-pen collagen shares some features with vertebrate type V collagen. Rotary shadowed images of acid-soluble collagen molecules extracted from beta-APN treated animals, positive staining of segment-long-spacing crystallites precipitated from pepsinized collagen, Western blots of the pepsinized alpha1 and alpha2 chains with antibodies to vertebrate types I, III and V collagens, and in situ gold immunolabeling of ECM collagen fibrils were examined. Our results showed that the tissue form of the sea-pen collagen is a 340-nm threadlike molecule, which is close to the vertebrate type V collagen with its voluminous terminal globular domain, the distribution of most of its polar amino-acid residues, and its antigenic properties.

Amino Acid Sequence↗
PubMed1992

Genetic variation and evolution of human parainfluenza virus type 1 hemagglutinin neuraminidase: analysis of 12 clinical isolates.

The extent of genetic variation and evolution in a population of human parainfluenza virus type 1 was investigated. The hemagglutinin neuraminidase genes of 13 isolates collected over a 26-year period were sequenced and compared. All isolates except the 1957 type strain were from a single geographic location and demonstrated significant consistent genetic change from the type strain (47/7 [nucleotide/amino acid] substitutions). Antigenic subgroup A isolates demonstrated minor intragroup differences (9/1 substitutions). However, 18/7 unique substitutions separated subgroup A from B regardless of geographic location or year of isolation. Multiple strains of both subgroups appeared and reappeared over decades with only minor variation. There may be significant genetic differences between clinical isolates based on geographic location, and progressive mutational change may occur. Previously defined antigenic and now genetic subgroups were stable and at least regional in distribution over the period studied. The biologic implications and extent of this variation need further evaluation.

Animals↗