Extensive and interrelated subcortical white and gray matter alterations in preterm-born adults.

Preterm birth is a leading cause for impaired neurocognitive development with an increased risk for persistent cognitive deficits in adulthood. In newborns, preterm birth is associated with interrelated white matter (WM) alterations and deep gray matter (GM) loss; however, little is known about the persistence and relevance of these subcortical brain changes. We tested the hypothesis that the pattern of correspondent subcortical WM and GM changes is present in preterm-born adults and has a brain-injury-like nature, i.e., it predicts lowered general cognitive performance. Eighty-five preterm-born and 69 matched term-born adults were assessed by diffusion- and T1-weighted MRI and cognitive testing. Main outcome measures were fractional anisotropy of water diffusion for WM property, GM volume for GM property, and full-scale IQ for cognitive performance. In preterm-born adults, reduced fractional anisotropy was widely distributed ranging from cerebellum to brainstem to hemispheres. GM volume was reduced in the thalamus, striatum, temporal cortices, and increased in the cingulate cortices. Fractional anisotropy reductions were specifically associated with GM loss in thalamus and striatum, with correlation patterns for both regions extensively overlapping in the WM of brainstem and hemispheres. For overlap regions, fractional anisotropy was positively related with both gestational age and full-scale IQ. Results provide evidence for extensive, interrelated, and adverse WM and GM subcortical changes in preterm-born adults. Data suggest persistent brain-injury-like changes of subcortical-cortical connectivity after preterm delivery.

The implementation of bedside bladder ultrasound technology: effects on patient and cost postoperative outcomes in tertiary care.

RESEARCH PROBLEM: The purpose of this study was to determine the effect of ultrasound assessment of bladder volume on patient and cost outcomes for patients needing postoperative catheterization. RESEARCH METHODS: Prospective descriptive and quasi-experimental designs were used with patients having general surgery (N = 50) and patients having orthopaedic surgery (N = 103), respectively. Four outcomes were measured: number of catheterizations over the hospitalization period, infection rates, cost, and patient and provider satisfaction. RESULTS: Accuracy of the technology ranged from .76 to .97 (p = .01). Catheterizations were avoided in 38% (in the "Due to Void" category) and in 81% (in the "Void with Residual" category) of patients in the general surgery group. Patients in the orthopaedic surgery group having ultrasound experienced 1.68 and the standard catheterization group, 1.96 catheterizations, throughout hospitalization. This difference was not statistically significant. UTI rates from admission to 30 days after discharge were 4% and 13% of patients in the ultrasound and standard orthopaedic groups, respectively, and 17% of patients in the general surgical population. Based on acquisition catheter cost, approximately 3 years of ultrasound machine use would be needed to recover the acquisition cost for each machine. The satisfaction rate with the technology was 93% of patients and 97% of nurses. In postoperative patients, bedside bladder ultrasound is accurate, is effective in decreasing numbers of catheterizations, reduces cost over time, and provides high patient and provider satisfaction.

Pupil perimetry using M-sequence stimulation technique.

PURPOSE: M-sequence stimulation technique allows mapping of the retinal function by multifocal electroretinographic (ERG) recordings. However, the information provided about visual field is limited to retinal function. Optic nerve diseases and diseases of the higher visual pathways usually show normal multifocal ERGs. Using pupillary responses instead of the electrical retinal responses might enhance the diagnostic possibilities of this system. The problems of local ERG recordings are very similar to those encountered in pupil perimetry: Local stimuli have to be dim to avoid or at least reduce stray-light responses. Dim stimuli, close to the absolute threshold, elicit only subtle pupillomotor responses. Therefore, techniques that are able to detect small focal responses are promising. METHODS: Pupillography was done by means of an infrared video camera and real time image processing (50 Hz) using a custom-designed videoboard in a personal computer (486). Recording conditions: The stimulus was presented on a monitor (75 Hz) in 26 cm distance from the patient's eyes. It contained 37 hexagons in a 25 degrees visual field. Each element changed between black (1.6 cd/m2) and white (160 cd/m2) after a binary M-sequence independently from other elements. Four thousand ninety six different stimulus pictures of 120-msec duration were shown during a single pupillogram recording. Thirty-seven local pupillograms were calculated in a cross-correlation of stimulus sequence and the pupil diameter. RESULTS: The pupillomotor fields in normals showed a shape and sensitivity distribution as known from conventional pupil perimetry techniques. Artificial paracentral scotomas (5 degrees) created by masking different locations could be demonstrated convincingly. Even in patients with optic nerve lesions it was possible to demonstrate visual field defects. CONCLUSIONS: Pupil perimetry using the M-sequence technique is a promising method of objective perimetry that may find its entrance into clinical application.

Improving pain management after total joint replacement surgery.

PURPOSE: To test the effects of implementing evidence-based postoperative pain management content and strategies on patient, provider (nurse and physician), and fiscal outcomes. SAMPLE: 118 patients, 57 before and 61 after implementation, having total knee replacement (TKR) (54%) and total hip replacement (THR) (45%), and 28 orthopaedic nurses. METHODS: A research utilization approach with a pretest/posttest design was used. Independent variables (interventions) included evidence-based pain management content, education of care providers and patients, and system changes at the point of care. Dependent variables (outcomes) were patient perception of the postoperative pain experience, provider practice patterns, and length of stay (LOS). FINDINGS: The hypotheses of decreased provider use of meperidine and increased use of hydromorphone, i.v. route, pain plans of care, and nurse knowledge were supported. LOS was significantly reduced. The patient hypotheses decreased pain intensity and side effects and increased satisfaction and function were not supported. CONCLUSION: Methodical implementation of evidence-based pain management information changed practice and fiscal outcomes. Improvement in the patient perception of pain management was more difficult to achieve.

The roles of the bacteriophage T4 r genes in lysis inhibition and fine-structure genetics: a new perspective.

Seldom has the study of a set of genes contributed more to our understanding of molecular genetics than has the characterization of the rapid-lysis genes of bacteriophage T4. For example, T4 rII mutants were used to define gene structure and mutagen effects at the molecular level and to help unravel the genetic code. The large-plaque morphology of these mutants reflects a block in expressing lysis inhibition (LIN), the ability to delay lysis for several hours in response to sensing external related phages attacking the cell, which is a unique and highly adaptive attribute of the T4 family of phages. However, surprisingly little is known about the mechanism of LIN, or how the various r genes affect its expression. Here, we review the extensive old literature about the r genes and the lysis process and try to sort out the major players affecting lysis inhibition. We confirm that superinfection can induce lysis inhibition even while infected cells are lysing, suggesting that the signal response is virtually instantaneous and thus probably the result of post-translational regulation. We identify the rI gene as ORF tk.-2, based on sequence analysis of canonical rI mutants. The rI gene encodes a peptide of 97 amino acids (Mr = 11.1 kD; pI = 4.8) that probably is secreted into the periplasmic space. This gene is widely conserved among T-even phage. We then present a model for LIN, postulating that rI is largely responsible for regulating the gpt holin protein in response to superinfection. The evidence suggests that the rIIA and B genes are not directly involved in lysis inhibition; rather, when they are absent, an alternate pathway for lysis develops which depends on the presence of genes from any of several possible prophages and is not sensitive to lysis inhibition.

[Continuous intestinal suture with polydioxanone]. / Fortlaufende Darmnaht mit Polydioxanon.

Within five years a total of 264 anastomoses were performed on 241 patients in a continuous extramucosal single-layer end-on suture technique with polydioxanone. In 133 small bowel, and 131 colonic anastomoses there were no leakages as the only complications attributable to the procedure. Both anastomotic failures occurred after colonic resections, for a 1.5% dehiscence rate. Small bowel anastomoses healed without complications.

[Ultra-high dosage lysis of deep venous thrombosis in ipsipedal administration along with compression]. / UHSK-Lyse tiefer Venenthrombosen bei ipsipedaler Applikation unter Kompressionsbedingungen.

The ipsipedal application of UHSK leads to a decisive increase of recanalisation. The fibrinolyticum will be brought in a greater quantity to the thrombotic region. The adjuvant supramalleolar compression and the increase of volume seems to be beneficial. The promising results of thrombolysis at elder thrombosis justify a late fibrinolysis, which is profitable too, for patients with early postoperative thrombosis at the end of the first wound healing phase.

Role of MgATP in the activation and reassociation of cAMP-dependent protein kinase I: consequences of replacing the essential arginine in cAMP binding site A.

The type I form of cAMP-dependent protein kinase binds MgATP with a high affinity, and binding of MgATP decreases the affinity of the holoenzyme for cAMP [Hofmann et al. (1975) J. Biol. Chem. 250, 7795]. Holoenzyme was formed here with a mutant form of the bovine recombinant type I regulatory subunit where the essential arginine in site A, Arg-209, was replaced with Lys. Although this mutation does not significantly change the high-affinity binding of MgATP to the holoenzyme, it does abolish high-affinity binding of cAMP to site A. In the absence of MgATP, binding of cAMP to site B is sufficient to promote dissociation of the holoenzyme complex and activation of the catalytic subunit [Bubis et al. (1988) J. Biol. Chem. 263, 9668]. In the presence of MgATP however, holoenzyme formed with this mutant regulatory subunit is very resistant to cAMP. The Kd(cAMP) was greater than 1 microM, and the Ka(cAMP) increased 60-fold from 130 nM to 6.5 microM in the presence of MgATP. Thus, MgATP serves as a lock that selectively stabilizes the holoenzyme and inhibits activation. Both site A and site B are shielded from cAMP in the presence of MgATP. These results suggest that Arg-209 may play a role in stabilizing the MgATP.holoenzyme complex in addition to its role in binding the exocyclic oxygens of cAMP when cAMP is bound to the regulatory subunit. The catalytic subunit also reassociates rapidly with this mutant regulatory subunit, and in contrast to the wild-type regulatory subunit, holoenzyme formation does not require MgATP.

A point mutation abolishes binding of cAMP to site A in the regulatory subunit of cAMP-dependent protein kinase.

Each regulatory subunit of cAMP-dependent protein kinase has two tandem cAMP-binding sites, A and B, at the carboxyl terminus. Based on sequence homologies with the cAMP-binding domain of the Escherichia coli catabolite gene activator protein, a model has been constructed for each cAMP-binding domain. Two of the conserved features of each cAMP-binding site are an arginine and a glutamic acid which interact with the negatively charged phosphate and with the 2'-OH on the ribose ring, respectively. In the type I regulatory subunit, this arginine in cAMP binding site A is Arg-209. Recombinant DNA techniques have been used to change this arginine to a lysine. The resulting protein binds cAMP with a high affinity and associates with the catalytic subunit to form holoenzyme. The mutant holoenzyme also is activated by cAMP. However, the mutant R-subunit binds only 1 mol of cAMP/R-monomer. Photoaffinity labeling confirmed that the mutant R-subunit has only one functional cAMP-binding site. In contrast to the native R-subunit which is labeled at Trp-260 and Tyr-371 by 8-N3cAMP, the mutant R-subunit is convalently modified at a single site, Tyr-371, which correlates with a functional cAMP-binding site B. The lack of functional cAMP-binding site A also was confirmed by activating the mutant holoenzyme with analogs of cAMP which have a high specificity for either site A or site B. 8-NH2-methyl cAMP which preferentially binds to site B was similar to cAMP in its ability to activate both mutant and wild type holoenzyme whereas N6-monobutyryl cAMP, a site A-specific analog, was a very poor activator of the mutant holoenzyme. The results support the conclusions that 1) Arg-209 is essential for cAMP binding to site A and 2) cAMP binding to domain A is not essential for dissociation of the mutant holoenzyme.

Stability of wild-type and mutant RTEM-1 beta-lactamases: effect of the disulfide bond.

Uniquely among class A beta-lactamases, the RTEM-1 and RTEM-2 enzymes contain a single disulfide bond between Cys 77 and Cys 123. To study the possible role of this naturally occurring disulfide in stabilizing RTEM-1 beta-lactamase and its mutants at residue 71, this bond was removed by introducing a Cys 77----Ser mutation. Both the wild-type enzyme and the single mutant Cys 77----Ser confer the same high levels of resistance to ampicillin in vivo to Escherichia coli; at 30 degrees C the specific activity of purified Cys 77----Ser mutant is also the same as that of the wild-type enzyme. Also, neither wild-type enzyme nor the Cys 77----Ser mutant is inactivated by brief exposure to p-hydroxymercuribenzoate. However, above 40 degrees C the mutant enzyme is less stable than wild-type enzyme. After introduction of the Cys 77----Ser mutation, none of the double mutants (containing the second mutations at residue 71) confer resistance to ampicillin in vivo at 37 degrees C; proteins with Ala, Val, Leu, Ile, Met, Pro, His, Cys, and Ser at residue 71 confer low levels of resistance to ampicillin in vivo at 30 degrees C. The use of electrophoretic blots stained with antibodies against beta-lactamase to analyze the relative quantities of mutant proteins in whole-cell extracts of E. coli suggests that all 19 of the doubly mutant enzymes are proteolyzed much more readily than their singly mutant analogues (at Thr 71) that contain a disulfide bond.(ABSTRACT TRUNCATED AT 250 WORDS)

Active-site mutants of beta-lactamase: use of an inactive double mutant to study requirements for catalysis.

We have studied the catalytic activity and some other properties of mutants of Escherichia coli plasmid-encoded RTEM beta-lactamase (EC 3.5.2.6) with all combinations of serine and threonine residues at the active-site positions 70 and 71. (All natural beta-lactamases have conserved serine-70 and threonine-71.) From the inactive double mutant Ser-70----Thr, Thr-71----Ser [Dalbadie-McFarland, G., Cohen, L. W., Riggs, A. D., Morin, C., Itakura, K., & Richards, J. H. (1982) Proc. Natl. Acad. Sci. U.S.A. 79, 6409-6413], an active revertant, Thr-71----Ser (i.e., residue 70 in the double mutant had changed from threonine to the serine conserved at position 70 in the wild-type enzyme), was isolated by an approach that allows identification of active revertants in the absence of a background of wild-type enzyme. This mutant (Thr-71----Ser) has about 15% of the catalytic activity of wild-type beta-lactamase. The other possible mutant involving serine and threonine residues at positions 70 and 71 (Ser-70----Thr) shows no catalytic activity. The primary nucleophiles of a serine or a cysteine residue [Sigal, I. S., Harwood, B. G., & Arentzen, R. (1982) Proc. Natl. Acad. Sci. U.S.A. 79, 7157-7160] at position 70 thus seem essential for enzymatic activity. Compared to wild-type enzyme, all three mutants show significantly reduced resistance to proteolysis; for the active revertant (Thr-71----Ser), we have also observed reduced thermal stability and reduced resistance to denaturation by urea.

[Prevention of postoperative Candida endomycoses. Candida colonization and Candida antibodies in patients undergoing cardiovascular surgery]. / Untersuchungen zur Prävention postoperativer Candida-Endomykosen. Candida-Besiedelung und Candida-Antikörper bei Patienten mit herz- und gefässchirurgischen Eingriffen.

Of 23 patients undergoing heart surgery and 11 patients undergoing vessel surgery 26 (76%) are colonized with Candida albicans on admission to hospital. Postoperatively the yeast quantity in the mucocutaneous flora remains nearly constant in both groups. However, during the 2nd-3rd week after operation 21 heart surgery patients and 3 vessel surgery patients developed a significant rise in Candida antibodies in the Ig-classes G, M and/or A. Eight weeks after surgery the IgM-antibodies once again reached preoperative values in 14 of 16 patients; the IgG- and IgA-antibodies, however, tend to be of longer persistent. The antibody response is most probably due to Candida invasion via tracheal traumatization by prolonged intubation. Persistent elevation of IgM-antibodies indicates preventive antifungal therapy.

Patch enlargement of the aortic and mitral valve rings with aortic and mitral double valve replacement. Experimental study.

The experimental results of patch enlargement of the aortic and mitral valve rings with aortic and mitral double valve replacement are reported. The operative technique of this new surgical method is described and the indications are discussed.

Binding properties of purified adult and fetal bovine serum albumin.

Fetal and adult bovine serum albumin were purified by standard techniques and then treated with activated charcoal at pH 2.75 to remove small molecular weight legands. The two proteins were compared and found to be identical by the following criteria: isoelectric focusing, electrophoresis on cellulose acetate and polyacrylamide gel, by the binding of bromcresol green, salicylic acid, diphenylhydantoin, and six different fatty acids (palmitic, stearic, oleic, elaidic, linoleic, and arachidonic). Reports of marked differences in the capacities of fatal and ault plasma to bind bilirubin and various drugs have usually been attributed to the presence of a "fetal" albumin. Our results, however, are consistent with the view that there is no difference between adult and fetal albumin and that the observed altered binding properties of fetal plasma are due to the tight binding of a ligand to fetal albumin in blood, which modifies the protein's affinity for lipophilic compounds.