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OBJECTIVE: This study was performed to determine the feasibility of measuring the elastic properties of the arterial wall in vivo. To prove this concept, elastic parameters were calculated from an aortic model of elastic behavior similar to a human aorta using computed tomography angiography (CTA) images. METHODS: We first constructed an aortic model from polydimethylsiloxane (PDMS). This model was inserted into a pulsatile flow loop. The model was then placed inside a computed tomography scanner. To estimate the elasticity values, we measured the cross-sectional area and the pressure changes in the model during each phase of the simulated cardiac cycle. A discrete wavelet transform (DWT) algorithm was applied to the CTA data to calculate the geometric changes in the pulsatile model over a simulated cardiac cycle for various pulsatile rates and elasticity values of the PDMS material. The elastic modulus of the aortic model wall was derived from these geometric changes. The elastic moduli derived from the CTA data were compared with those obtained by testing strips of the same PDMS material in a tensile testing machine. Our two aortic models had elastic values at both extremes of those found in normal human aortas. RESULTS: The results show a good comparison between the elastic values derived from the CTA data and those obtained in a tensile testing machine. In addition, the elasticity values were found to be independent of the pulsatile rate for mixing ratios of 6:1 and 9:1 (p = .12 and p = .22, respectively). CONCLUSIONS: The elastic modulus of a pulsatile aortic model may be measured by electrocardiographically-gated multi-detector CTA protocol. This preliminary study suggests the possibility of determining non-invasively the elastic properties of a living, functioning aorta using CTA data.

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BACKGROUND: Different stress-strain definitions are used in the literature to measure the elastic modulus in aortic tissue. There is no agreement as to which stress-strain definition should be implemented. The purpose of this study is to show how different results are given by the various definitions of stress-strain used and to recommend a specific definition when testing aortic tissues. METHODS: Circumferential specimens from three patients with ascending thoracic aortic aneurysm (ATAA) were obtained from the greater curvature and their tensile properties were tested uniaxially. Three stress definitions (second Piola-Kirchhoff stress, engineering stress and true stress) and four strain definitions (Almansi-Hamel strain, Green-St. Venant strain, engineering strain and true strain) were used to determine the elastic modulus. RESULTS: We found that the Almansi-Hamel strain definition exhibited the highest non-linear stress-strain relation and consequently may overestimate the elastic modulus when using different stress definitions (second Piola-Kirchhoff stress, engineering stress and true stress). The Green-St. Venant strain definition yielded the lowest non-linear stress-strain relation using different definitions of stress, which may underestimate the values of elastic modulus. Engineering stress and strain definitions are only valid for small strains and displacements, which make them impractical when analysing soft tissues. We show that the effect of varying the stress definition on the elastic modulus measurements is significant for maximum elastic modulus but not when calculating the hypertensive elastic modulus. CONCLUSIONS: It is important to consider which stress-strain definition is employed when analysing soft tissues. Although the true stress-true strain definition exhibits a non-linear relation, we favour it in tissue mechanics because it gives more accurate measurements of the material's response using the instantaneous values.

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OBJECTIVE: Ascending thoracic aortic aneurysms (ATAA) are a life-threatening condition due to the risk of rupture or dissection. This risk is increased in the presence of a bicuspid aortic valve (BAV). The purpose of this study was to provide data on the elastic modulus of aortic wall of ATAA using uniaxial tensile testing in two different areas of the stress-strain relationship: physiological and maximum range of stresses. The influence of tissue location, tissue orientation and valve type on these parameters was investigated. MATERIALS AND METHODS: Tissues freshly excised from ATAA with bicuspid or tricuspid aortic valve were obtained from greater and lesser curvature (GC and LC) and the specimens were tested uniaxially in circumferential (CIRC) and longitudinal (LONG) orientation. Maximum elastic modulus (MEM) was given by the maximum slope of the stress-strain curve before failure. Physiological modulus (PM) was derived from the Laplace law and from ranges of pressure of 80-120 mmHg. Means of each group of specimen were compared using Student's t-test to assess the influence of location, orientation and valve type on each mechanical parameter. RESULTS: PM was found to be significantly lower than the MEM (p < 0.001). The MEM and PM were significantly higher (p < 0.01) in the CIRC (n = 66) than in the LONG orientation (n = 42). The MEM was higher in the circumferential orientation in the BAV group (p < 0.001 in GC and p < 0.05 in LC). MEM and PM in GC specimens were higher in the longitudinal orientation than the LC specimens (p < 0.05). CONCLUSION: This study demonstrates the anisotropy of the aortic wall in ATAA and provides data on the mechanical behaviour in the physiological range of pressure.

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Recently, we have reported that cell wall pectins are internalized into apical meristem root cells. In cells exposed to the fungal metabolite brefeldin A, all secretory pathways were inhibited, while endocytic pathways remained intact, resulting in accumulation of internalized cell wall pectins within brefeldin A-induced compartments. Here we report that, in addition to the already published cell wall epitopes, rhamnogalacturonan I and xyloglucans also undergo large-scale internalization into dividing root cells. Interestingly, multilamellar endosomes were identified as compartments internalizing arabinan cell wall pectins reactive to the 6D7 antibody, while large vacuole-like endosomes internalized homogalacturonans reactive to the 2F4 antibody. As all endosomes belong topographically to the exocellular space, cell wall pectins deposited in these "cell wall islands", enclosed by the plasma-membrane-derived membrane, are ideally suited to act as temporary stores for rapid formation of cell wall and generation of new plasma membrane. In accordance with this notion, we report that all cell wall pectins and xyloglucans that internalize into endosomes are highly enriched within cytokinetic cell plates and accumulate within brefeldin A compartments. On the other hand, only small amounts of the pectins reactive to the JIM7 antibody, which are produced in the Golgi apparatus, localize to cell plates and they do not accumulate within brefeldin A compartments. In conclusion, meristematic root cells have developed pathways for internalization and recycling of cell wall molecules which are relevant for plant-specific cytokinesis.

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We have surveyed the publicly available genome sequence of Corynebacterium diphtheriae (www.sanger.ac.uk) to identify components of the phosphotransferase system (PTS), which plays a central role in carbon metabolism in many bacteria. Three gene loci were found to contain putative pts genes. These comprise: (i) the genes of the general phosphotransferases enzyme I (ptsI) and HPr (ptsH), a fructose-specific enzyme IIABC permease (fruA), and a fructose 1-phosphate kinase (fruK); (ii) a gene that encodes an enzyme IIAB of the fructose/mannitol family, and a novel HPr-like gene, ptsF, that encodes an HPr domain fused to a domain of unknown function; (iii) and a gene for a glucose-specific enzyme IIBCA (ptsG). A search for genes that may be putative PTS-targets or that may operate in general carbon regulation revealed a possible regulatory gene encoding an antiterminator protein downstream from ptsG. Furthermore, genes were detected encoding glycerol kinase, glucose kinase, and a homologue of the global activator of carbon catabolite repression in Escherichia coli, CAP. The possible significance of these observations in carbon metabolism and the novel features of the detected genes are discussed.

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The toxicity of 10 chemicals, including pesticides (carbaryl, chlordane, heptachlor, and triadimefon), solvents (carbon tetrachloride, dichloromethane, tetrachloroethylene, and trichloroethylene), and industrial chemicals [diethylhexylphthalate (DEHP) and phenol] was examined in the liver, kidneys, spleen, thymus, and adrenals of female F344 rats after 1 or 14 d of oral dosing. For each chemical, 4 doses were based on fractions of the acute LD50, which was estimated using an abbreviated (up-and-down) method. A multivariate analysis (MANOVA) was conducted for each organ using selected measures of toxicity. A post hoc contrast analysis was also conducted for significant MANOVA results in order to determine effective and ineffective doses. A single dose of heptachlor resulted in necrotic lymphocytes in the spleen and thymus at doses > or = 23 mg/kg. Triadimefon altered liver and spleen weights; this effect has not been described previously. Dichloromethane (> or = 337 mg/kg/d for 14 d) increased the incidence of necrosis of individual centrilobular hepatocytes. Trichloroethylene-induced hepatotoxicity was obtained at doses an order of magnitude lower than those reported in the literature. Acute DEHP (150 mg/kg) increased mitotic figures in hepatocytes, which were replaced by hepatocellular cytomegaly after 14 d of dosing at the same level. Following phenol exposure, there was an increased incidence in hepatocellular necrosis at 1 d, and an increased incidence of kidney lesions at 1 and 14 d; these findings were considered to be the result of vascular stasis. Overall, the algorithm used to select doses was effective for both 1- or 14-d dosing regimens. For all chemicals except carbon tetrachloride, the lowest effective dose for systemic toxicity was within the range of 3-56% of the LD50 for acute dosing, and 1-30% of the LD50 for repeated administration.

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Toxicity data collected under standardized test conditions may be of the utmost importance in health risk assessment, in which human exposure limits are often derived from laboratory experiments. A standardized approach to data collection is also important for evaluating the sensitivity and specificity of test methods used to determine toxic potential. Several experiments were undertaken to determine the effects of chemical exposures using a multidisciplinary screening battery, which included tests for systemic, neurological and developmental toxicity. The effects of 1- and 14-d exposures to 10 chemicals on systemic and neurological indices of toxicity were determined in female F344 rats using standardized test batteries. Parallel experiments determined chemical effects on prenatal and postnatal development following exposure of the dams for 14 d. The chemicals included four pesticides (carbaryl, triadimefon, chlordane, and heptachlor), four solvents (trichloroethylene, tetrachloroethylene, carbon tetrachloride, and dichloromethane), and two industrial compounds (phenol and diethylhexyl phthalate). The results showed that the chemicals produced markedly different qualitative patterns of effect on systemic, neurological, and developmental indices of toxicity. Differences in the pattern of systemic and neurological effects were also obtained that depended on dosing duration. Quantitative analyses indicated that the highest ineffective dose as well as the lowest effective dose could vary by as much as two orders of magnitude across the different indices of toxicity. These results clearly show that a test battery focused on a single endpoint of toxicity cannot be used to accurately predict either qualitatively or quantitatively a chemical's systemic, neurological, and developmental toxicity profile.

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In 128 patients (20 f., 98 m., age between 26 and 88 years, mean age 59 years) with acute myocardial infarction, infarct size was calculated from CK and MBCK serum activity using the individual fractional decay rate kD. Only a moderate correlation could be found between infarct size calculated from CK-serum curves (CK-IG) and from MBCK (MBCK-IG) (r = 0.65). Only little improvement was achieved by excluding those patients who had been resuscitated (n = 13, r = 0.69). In 20% of the patients there was a good correlation (+/- 5 g eq) between CK-IG and MBCK-IG. In 14% MBCK-IG was larger and in 66% smaller than CK-IG. Calculation of MBCK-IG is based on a constant MBCK percentage of CK in myocardium, namely 14%, and assumes that the distribution volume and amount of enzyme released into the serum is the same for CK and MBCK. Thus the percentage of MBCK in serum compared to CK should also be 14%. We found an average value approaching this (13.6%), but with a wide range between 4.7 and 21.2%. It is this variation which is responsible for the poor correlation between both IGs.

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The purification of the Escherichia coli dnaB protein by affinity chromatography on nucleotides bound to agarose is described. The dnaB protein, which contains an associated ribonucleoside triphosphatase activity (Wickner, S., Wright, M., and Hurwitz, J. (1974) Proc. Natl. Acad. Sci. U. S. A. 71, 783-787) binds to immobilized ATP, ADP, and UDP, but not to AMP. The type of linkage of ATP to agarose influences the adsorption, elution, and purification of the enzyme. Optimal purification is achieved using ATP bound to agarose via its oxidized ribose moiety. By this means, the dnaB protein can be obtained at least 95% electrophoretically pure after only three purification steps. The enzyme can be eluted from immobilized nucleoside-5'-di- and -triphosphates by ATP, ADP, and pyrophosphate, but not by AMP or orthophosphate. ADP and pyrophosphate, as well as the substrate ATP in high concentration are at the same time inhibitors of the ribonucleoside triphosphatase. The dnaB complementing and ribonucleoside triphosphatase activities could not be separated from each other by affinity chromatography, supporting the finding of others that they both reside on the same protein complex, namely a dnaB multimer. The results indicate that the dnaB protein binds to immobilized nucleotides by means of its ribonucleoside triphosphatase, and that at least the pyrophosphate moiety is essential for adsorption as well as elution of the enzyme.

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An Escherichia coli K12 dnaB dnaC mutant was constructed by P1 transduction of the dnaC allele into a dnaB recipient stain. dnaB dnaC transductant were discriminated from dnaB mutants by their inability to grow at 40 degree C after lysogenization with phage P1bac. The dnaB dnaC mutant character was verified by 1. P1 transduction, and 2. by in vitro complementation with dnaB and dnaC wild type protein fractions. DNA synthesis was studied in strains containing dnaB, dnaB dnaC alleles in an otherwise uniform genetic background with the dnaB character either unsuppressed or suppressed by P1bac prophage. Degradation at 42 degree C of [3H]-thymidine pulse-labeled DNA in dnaB and dnaB dnaC mutants is suppressed by P1bac. However, unlike the dnaC mutant, the P1bac lysogen of the dnaB dnaC mutant exhibits an abrupt cessation of DNA synthesis and less residual cell divisions at 42 degree C indicating an inhibition of DNA chain elongation rather than a defect in DNA initiation. It is suggested that denaturation of the dnaB protein effects the dnaC function.

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Mutagenized E. coli B/r cells were subjected to a procedure designed to select mutants temperature-sensitive for initiation of deoxyribonucleic acid (DNA) replication. Seventeen mutants exhibiting limited residual DNA synthesis at 42 C were obtained and the dna(-) sites were mapped genetically. Sixteen of the sites map near dnaA, dnaB, and dnaC. One mutant (dna-208) maps in a new location between the trp and his genes. We propose to call this mutant dnaI208. In complementation experiments dnaC(+) and dnaI(+) were dominant to dnaC(-) and dnaI(-) alleles, respectively. However, dnaA(-) was dominant to the wild-type allele dnaA(+). All dnaA mutants and four out of six dnaC mutants could be suppressed by F factor integration. The pattern of suppression was specific for each mutant.

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High-temperature treatment of thermosensitive dna mutants lysogenic for phage lambda leads to prophage induction and release of phage (at the permissive temperature) in elongation-defective mutants of the genotypes dnaB, dnaE, and dnaG. In initiation-defective mutants no prophage induction occurs at 42 C in mutants of the genotype dnaA, whereas with a dnaC mutant as well as with strain HfrH 252 (map position not yet known) phages are released at 42 C. DNA degradation at the replication fork at 42 C is observed in all dnaB(lambda) mutants tested, but not in mutants of the genotypes dnaE(lambda) and dnaG(lambda). Therefore, degradation of replication fork DNA is not a prerequisite for prophage induction.

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