[Decrease of the operative risk in reinterventions on valve prostheses].
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Aortic Valve↗
Biomedical subjects
J Swanson
Publications and source records attributed to J Swanson.
Gene conversion involving the pilin structural gene correlates with pilus+ in equilibrium with pilus- changes in Neisseria gonorrhoeae.
Gonococci (Gc) exhibit pilus+----pilus- "phase transitions" at high frequency, but only some of the pilus- Gc can revert to pilus+ phenotype. We examined reversible phase transitions between pilus+ Gc and a particular pilus- variant (P-rp+ phenotype) whose pilin mRNA carries a unique block of nucleotides encoding an "assembly missense" pilin polypeptide. The results show that Gc pilus+ in equilibrium with P-rp+ transitions can result from intragenic recombination in which there is nonreciprocal exchange of partially homologous DNA sequences from a partial pilin gene (in silent, storage form) into the expression locus' complete pilin gene. Hence Gc pilus phase variation, like pilus antigenic variation, can occur by gene conversion of the pilin structural gene expression locus.
Bacterial Adhesion↗
Effect of alterations in the size of the vacuolar compartment on pinocytosis in J774.2 macrophages.
J774.2 macrophages cultured in medium containing 10 mg/ml sucrose accumulate the sugar by pinocytosis and become highly vacuolated, due to the sugar's osmotic effect within the vacuolar compartment. When such cells are incubated in medium containing 0.5 mg/ml invertase, the enzyme reaches the sucrose vacuoles by pinocytosis, then cleaves the sugar to more permeant monosaccharides. Within 4 hours, the vacuoles shrink to smaller, phase-dense organelles (Cohn and Ehrenreich, 1969, J. Exp. Med., 129:201). We have used this reversible expansion of the lysosomal compartment to address two questions: (1) Does the increased size of the lysosomal compartment affect pinocytic accumulation of solute, and (2) what is the fate of the vacuolar membrane and its soluble content during invertase-induced vacuole shrinkage? Using lucifer yellow (LY) as a probe for pinocytic fluid influx and efflux, we found that vacuolated cells accumulated 30-50% less LY than controls and returned to higher rates of pinocytosis after invertase-induced vacuole shrinkage. A similar reduction in LY accumulation was achieved after feeding cells latex beads to increase the size of the lysosomal compartment. Thus, treatments that increased the size of the lysosomal compartment reduced solute accumulation via pinocytosis. A dramatic shrinkage of LY-containing sucrose vacuoles followed pinocytosis of invertase. Despite this reduction in size of the LY-containing vacuoles, the overall rate of LY efflux did not increase significantly during invertase-induced vacuole collapse. Electron microscopy revealed that during shrinkage, the excess vacuolar membrane was compressed into whorled membranous organelles (residual bodies), with fluid markers (colloidal gold and, by inference, LY) trapped inside. The trapping of LY inside lysosomes as J774.2 macrophages returned to their normal dimensions indicates that nearly all of the surplus membrane contents were removed from circulation as well.
Cells, Cultured↗
Retrograde cardioplegia through the right atrium.
Retrograde cardioplegia through the coronary sinus has been shown to overcome the problems of cardioplegia delivery in aortic valve incompetence and coronary stenosis. Since specific complications, such as coronary sinus injury and lack of perfusion of the thebesian veins, may result from its use, we present a technique of cardioplegia delivery through the right atrium after caval and pulmonary artery exclusion that we believe is safer and more reliable than delivery through the coronary sinus.
Cardiopulmonary Bypass↗
Piliation control mechanisms in Neisseria gonorrhoeae.
Gonococci (Gc) undergo pilus+ to pilus- "phase transitions" readily in vitro. In the present study we sequenced pilin mRNA from reverting, pilus- Gc by oligonucleotide primer extension and compared these pilin mRNA sequences with those expressed by their pilus+ predecessors and pilus+ revertants. The results suggest that genetic rearrangement within the pilin structural gene can generate defective pilin gene products, resulting in a pilus- phenotype. These pilus- Gc give rise to pilus+ revertants upon reconstitution of their modified pilin gene.
Bacterial Outer Membrane Proteins↗
Patterns of sarcomere activation, temperature dependence, and effect of ryanodine in chemically skinned cardiac fibers.
Functionally skinned and electrochemically shunted myocytes were prepared by perfusing rat hearts with collagenase in order to obtain a technically improved measurement of sarcomere dynamics and to evaluate the role of sarcoplasmic reticulum in situ with respect to contractile activation. In the presence of micromolar calcium, the myocytes exhibited phasic and propagated contraction waves beginning at one end and proceeding along the myocyte. Beating rates, the propagation velocity of the activation wave, and single sarcomere shortening and relaxation velocities were obtained by manual or automated analysis of 16-mm film recorded at 170 frames/s from a camera attached to a microscope that was equipped with a temperature-controlled stage. In parallel experiments, calcium accumulation by the sarcoplasmic reticulum of the myocytes in situ was measured by direct isotopic tracer methods. The frequency (10-38 min-1) of spontaneous contractions, the velocity (1.9-7.4 microns . s-1) of sarcomere shortening, and the velocity (1.7-6.8 microns . s-1) of sarcomere relaxation displayed identical temperature dependences (Q10 = 2.2), which are similar to that of the calcium pump of sarcoplasmic reticulum and are consistent with a rate limit imposed by enzyme-catalyzed mechanisms on all these parameters. On the other hand, the velocity (77-159 microns . s-1) of sequential sarcomere activation displayed a lower temperature dependence (Q10 = 1.5), which is consistent with a diffusion-limited and self-propagating release of calcium from one sarcomere to the other. The phasic contractile activity of the dissociated myocytes was inhibited by 10(-8)-10(6) M ryanodine (and not by myolemmal calcium blockers) under conditions in which calcium accumulation by sarcoplasmic reticulum in situ was demonstrated to proceed optimally. The effect of ryanodine is attributed to an interaction of this drug with sarcotubular structures, producing inhibition of calcium release from the sarcoplasmic reticulum. The consequent lack of sarcomere activation underlines the role of sarcoplasmic reticulum uptake and release in the phasic contractile activation of the electrochemically shunted myocytes.
Alkaloids↗
Risk and consequence of infection at the site of microsurgical repair: an experimental model.
The consequences of postoperative infection at the site of microsurgical repair were studied. The experiment used 60 Wistar rats which were divided into 3 groups of 20 each. The femoral artery, vein, and nerve were transected unilaterally and repaired using microsurgical techniques. Staphylococcus aureus was then inoculated into the wound of group 1. Beta-hemolytic Streptococcus was inoculated into the wound of group 2. Group 3 rats served as controls. Clinical, anatomical, bacteriological and histopathological examinations were performed on postoperative days 8 and 30. Wounds in which S. aureus was inoculated demonstrated a significant tendency toward vascular thrombosis with extensive tissue destruction. In the majority of these cases, the necrosis caused by the primary infection engendered an associated, opportunistic infection. Streptococcal inoculations in group 2 demonstrated less severe changes than in group 1. These changes were characterized by injury of the vascular structures themselves with a significant tendency for thickening of the arterial wall, perivascular inflammation, and hematoma formation at the site of the repair. This model allows demonstration of the enzymatic and toxic consequences of bacterial infection in a postoperative site characterized by cellular destruction and interstitial edema surrounding foreign bodies represented by sutures.
Animals↗
Partial reversal of low molecular weight heparin (PK 10169) anti-Xa activity by protamine sulfate: in vitro and in vivo study during cardiac surgery with extracorporeal circulation.
Neutralization of a low molecular weight (LMW) heparin fraction by protamine sulfate was evaluated in vitro and in vivo. Anti-Xa and anti-IIa activities were measured by amidolytic and coagulation methods (activated partial thromboplastin time, APTT). Fifteen patients (4 males and 11 females) underwent surgery with extracorporeal circulation. In vitro, anti-Xa and anti-IIa activities and APTT of unfractionated heparin were neutralized with a protamine/heparin (P/H) gravimetric ratio of 1.6, 1.33 and about 2, respectively. Anti-IIa activity and APTT induced by PK 10169 were completely corrected at a P/H ratio of 1 and 2, respectively, while anti-Xa activity was incompletely neutralized at a ratio of 5. In vivo, in 9 patients who did not receive intravenous protamine sulfate, a good correlation was found between doses of PK 10169 infused, anti-IIa plasma level and blood loss. In 3 patients who were treated prophylactically with protamine, bleeding was normal or only slightly increased. In 3 patients who received protamine because of hemorrhage, mean anti-Xa and anti-IIa were 2.3 and 0.54 U before and 1.32-0.06 U after neutralization. Bleeding was stopped by a second dose of protamine in 1 patient, but blood loss was abnormal in the other patients. However, a correlation between bleeding and anti-Xa or anti-IIa activities was not clearly evident.
Adolescent↗
Presentation rate effects on paired associate learning by attention deficit disordered children.
2 methods of varying presentation rate during paired associate learning were contrasted in attention deficit disordered (ADD) children. Previous studies have varied presentation rate across different (fixed rate) lists, and they have demonstrated that ADD children perform poorly at slower rates. In the present study, this method of presentation was contrasted with one in which half the items within a single list were presented at a fast rate and half at a slow rate. The debilitating effect of the slow rate was obtained in ADD children (but not in normal controls) only with the fixed list method. This finding suggests that the rate effects occur in ADD children because they are vulnerable to the experimental context created when items are presented at a slow rate over an extended time period.
Attention↗
Pilus- gonococcal variants. Evidence for multiple forms of piliation control.
Pilus+ to pilus- transitions of gonococci (Gc) that involve rearrangement of pilin gene DNA yield the P-n phenotype, which is incapable of reversion (to pilus+). Reversion to pilus+ is found for nonpiliated Gc that have undergone no apparent pilin gene rearrangement. Among the reverting, nonpiliated Gc, two distinct phenotypes (P-rp- and P-rp+) occur and are differentiated according to their synthesis (or lack) of pilin subunits; both P-rp- and P-rp+ Gc contain pilin-specific mRNA. The occurrence of these different pilus- phenotypes strongly suggests that several mechanisms can account for changes in the piliation status of Gc; one of these involves pilin gene rearrangement but the others apparently operate at posttranscriptional levels. Reverting pilus- Gc may have a pathogenic advantage in being able to reversibly alter their host cell adherence-promoting surface properties through high frequency transitions in piliation status.
Bacterial Outer Membrane Proteins↗
Reversibility of muscular ischemia: a histochemical quantification by the nitroblue tetrazolium (NBT) test.
The degree of muscular ischemia and its reversibility can be quantified in the early stages. This histochemical enzymatic study utilized Nitroblue tetrazolium (NBT) which when reduced by tissue dehydrogenase produces a blue pigment: "formazan." Seventy Wistar rats were subjected to transient hindlimb ischemia by means of a tourniquet for 3, 6, 9, 12, 15 and 18 hours, followed by reperfusion. Microsurgical muscle biopsies were obtained in each rat at 1 and 12 hours, and 3, 7, 14 days after reperfusion. Time increased in muscle staining demonstrated a succino-dehydrogenase deficit confirmed by clinical and histopathological follow-up. NBT staining time was 2 minutes (+/- 8 sec.) in the control group, between 2 and 6 minutes in the reversible ischemia group (rats with 3 and 6 hours of tourniquet), and more than 9 minutes (+/- 14 sec.) in the irreversible ischemia group (animals with more than 9 hours of tourniquet). In vascular surgery and in limb reimplantation this protocol is a practical method of evaluating cytoplasmic enzymatic activity and the status of myofibrillar oxidation in the early phases of ischemic injury, before histologic changes are clearly delineated.
Animals↗
The delayed and saline wattle reactions in broilers challenged with bovine serum albumin.
Bovine serum albumin was used to study the development of a wattle reaction in broiler-type chickens. A strong response developed in both nonsensitized and sensitized chickens and the frequency of such was shown to be dependent on the method used for the sensitization. A similar swelling developed in a proportion of the wattles injected with saline and this too seemed to be influenced by the sensitization method. Moreover, both types of responses were shown to be somewhat unstable during the first 24 hr postchallenge.
Animals↗
Nucleotide sequence of the maize transposable element Mul.
A cloned DNA fragment from the maize allele Adhl-S3034 contains all of Mul, an insertion element involved in Robertson's Mutator activity. The element is 1367 base pairs (bp) long and is flanked by nine bp direct repeats of insertion site DNA. It has inverted terminal repeats of 215 and 213 bp showing 95% homology. Within the element are two direct repeats of 104 bp showing 96% homology. Four open reading frames (ORFs) were found, two in each DNA strand. Mul can be divided into two halves, each containing one terminal inverted repeat, an internal direct repeat, and two overlapping ORFs. The GC content of each half is high (70%), while that of a central 60 base portion of the element is low (26%). The central region contains the only sequence resembling the TAATA Goldberg and Hogness eukaryotic promoter signal. Multiple copies of DNA sequences related to Mul found in Mutator maize plants are generally similar in organization to the cloned element. A larger version containing a discrete 300 to 400 base pair insertion was found in some Mutator lines.
Base Sequence↗
DNA insertion in the first intron of maize Adh1 affects message levels: cloning of progenitor and mutant Adh1 alleles.
A genetically unstable mutant allele of maize alcohol dehydrogenase-1 (Adh1) causes reduced levels of messenger RNA. This mutant was derived from a Robertson's mutator genetic background and previously was shown to be associated with an approximately 1.5-kilobase insertion. This report compares a genomic clone of the allele, Adh1-S3034, with a clone of its progenitor nonmutant allele, Adh1-S. No rearrangements of either recombinant molecule occurred during the cloning process. The mutant differs from its progenitor allele by the insertion of 1.4 kilobases of new DNA. Sequence determination of the insertion borders localized it to intron-1, 73 base pairs downstream from the 5' exon/intron junction in the Adh1-S gene. Insertion of the 1.4-kilobase element was associated with a 9-base-pair direct duplication of intron sequence. We suspect that this insertion, Mu1, affects message levels by depressing RNA processing or transcription.
Alcohol Dehydrogenase↗
The effectiveness of prophylactic subcutaneous mastectomy in Sprague-Dawley rats induced with 7,12-dimethylbenzanthracene.
Fifty-day-old female, albino Sprague-Dawley rats were induced with DMBA, a potent carcinogen that reliably causes multiple breast tumors. Prophylactic subcutaneous mastectomy was performed 2 weeks after injection. Control animals had no breast tissue removed and developed 5.14 tumors per animal, or 1.3 tumors per quadrant. Animals subjected to 50 percent (unilateral) mastectomy developed 5.4 tumors per animal, with 0.72 tumors per surgical quadrant. Animals that underwent 75 percent (three quadrant) mastectomy developed 4.33 tumors per animal, or 0.48 tumors per surgical quadrant. The incidence of tumors was significantly reduced in the surgical areas. However, the risk of any animal developing a breast neoplasm was not reduced in direct proportion to the percentage of breast tissue removed. We believe that this model may prove valuable in evaluating prophylactic subcutaneous mastectomy.
9,10-Dimethyl-1,2-benzanthracene↗
Proteins IA and IB exhibit different surface exposures and orientations in the outer membranes of Neisseria gonorrhoeae.
Exposure of whole gonococci to proteinase K resulted in cleavage of protein I (P.I) of the organism in situ. P.I subunits in the P.IB group were cleaved into two membrane-associated fragments, whereas P.IA subunits were cleaved by proteinase K to yield a single membrane-associated fragment slightly smaller in apparent size than the intact P.IA subunit. These data suggest that P.IA and P.IB subunits are quite different in their surface exposures and orientations in the gonococcal outer membrane; P.IB subunits likely have both termini buried in the membrane, whereas P.IA subunits have one of their termini exposed on the surface of the organism.
Bacterial Outer Membrane Proteins↗
Bacteriuria screening by direct bioluminescence assay of ATP.
A direct bioluminescence assay for bacteriuria screening is described and compared with the MS-2 system (Abbott Laboratories, Irvine, Tex.) and the chemical strip, Gram smear, and calibrated-loop methods. A total of 973 specimens were tested. Unlike previously described bioluminescence methods, this test measures total ATP in urine without pretreatment of samples to remove somatic ATP. The result was compared with an ATP standard (20 ng/ml). A low result (less than 3% of standard) was interpreted as negative and a high result (greater than 10% of standard) as positive. Samples with intermediate results (38% of total) were incubated at 35 degrees C in thioglycolate broth (1:10). A 2% increase in ATP concentration was interpreted as positive. The sensitivity of this method for detecting greater than 10(5) pathogens per ml was 92.3% and was comparable to those of the MS-2 system (92.7%) and the Gram smear method (90.5%). The chemical strip method was less sensitive (84.0%). The direct bioluminescence method was more sensitive than were the MS-2 system and the Gram smear method for detecting low-level bacteriuria (less than 10(3) to 10(5) organisms per ml), primarily because of associated pyuria. Thioglycolate broth provided a suitable medium for ATP production, and 5% CO2 decreased bacterial ATP synthesis during log-phase growth. The direct bioluminescence assay is rapid, simple, cost-effective, and reliable for bacteriuria screening.
Adenosine Triphosphate↗
Gonococcal pilus subunit size heterogeneity correlates with transitions in colony piliation phenotype, not with changes in colony opacity.
The apparent subunit sizes for pili of gonococci (Gc) have been visualized by using either Iodogen 125I-labeled whole Gc or immunoblotting with antipilus antiserum. These methods permitted definition of pilus subunit sizes for Gc of a given strain that had undergone changes either in piliation phenotype or in colonial opacity/protein II phenotype. The results indicate that pilus subunit size does not change coincident with changes in colony opacity/protein II phenotypes; but change in pilus subunit size is seen after a change in piliation phenotype (P+ leads to P++, and vice versa). Marked diversity in pilus subunit sizes is found for Gc of individual strains when P+ derivatives of P- colonies are compared. This diversity extends to pilus subunits of Gc found in single colonies; two distinct pilus forms were demonstrated for Gc residing in several single colonies. These findings show that Gc of a given strain are able to express any of a number of different pilus subunit size forms.
Bacterial Proteins↗