Inhibition of the hammerhead ribozyme cleavage reaction by site-specific binding of Tb

Terbium(III) [Tb(III)] was shown to inhibit the hammerhead ribozyme by competing with a single magnesium(II) ion. X-ray crystallography revealed that the Tb(III) ion binds to a site adjacent to an essential guanosine in the catalytic core of the ribozyme, approximately 10 angstroms from the cleavage site. Synthetic modifications near this binding site yielded an RNA substrate that was resistant to Tb(III) binding and capable of being cleaved, even in the presence of up to 20 micromolar Tb(III). It is suggested that the magnesium(II) ion thought to bind at this site may act as a switch, affecting the conformational changes required to achieve the transition state.     

Single injection thermodilution. A flow-corrected method

BACKGROUND: Application of the Stewart-Hamilton equation in the thermodilution technique requires flow to be constant. In patients in whom ventilation of the lungs is controlled, flow modulations may occur leading to large errors in the estimation of mean cardiac output. METHODS: To eliminate these errors, a modified equation was developed. The resulting flow-corrected equation needs an additional measure of the relative changes of blood flow during the period of the dilution curve. Relative flow was computed from the pulmonary artery pressure with use of the pulse contour method. Measurements were obtained in 16 patients undergoing elective coronary artery bypass surgery. In 11 patients (group A), pulmonary artery pressure was measured with a catheter tip transducer, in a partially overlapping group of 11 patients (group B), it was measured with a fluid-filled system. For reference cardiac output we used the proven method of four uncorrected thermodilution estimates equally spread over the ventilatory cycle. RESULTS: A total of 208 cardiac output estimates was obtained in group A, and 228 in group B. In group B, 48 estimates could not be corrected because of insufficient pulmonary artery pressure waveform quality from the fluid-filled system. Individual uncorrected Stewart-Hamilton estimates showed a large variability with respect to their mean. In group A, mean cardiac output was 5.01 l/min with a standard deviation of 0.53 l/min, or 10.6%. After flow correction, this scatter decreased to 5.0% (P < 0.0001). With no bias, the corresponding limits of agreement decreased from +/- 1.06 to +/- 0.5 l/min after flow correction. In group B, the scatter decreased similarly and the limits of agreement also became +/- 0.5 l/min after flow correction. CONCLUSION: It was concluded that a single thermodilution cardiac output estimate using the flow-corrected equation is clinically feasible. This is obtained at the cost of a more complex computation and an extra pressure measurement, which often is already available. With this technique it is possible to reduce the fluid load to the patient considerably.     

Effects of bryostatin 1, a novel anticancer agent, on intestinal transport and barrier function: role of protein kinase C

BACKGROUND: Bryostatin 1 is a novel chemotherapeutic agent that activates specific members of the protein kinase C (PKC) family in a complex pattern overlapping with, but distinct from, that of tumor-promoting phorbol esters. Phorbol esters profoundly altered epithelial phenotype, abolishing both barrier function and Cl secretion (the latter due to loss of a key transport site, the Na-K-Cl cotransporter). The effects of bryostatin 1 on these parameters are unknown. METHODS: Cl secretion and barrier function of T84 human intestinal epithelia were assessed as cyclic adenosine monophosphate-stimulated short-circuit current and transepithelial resistance, respectively. Na-K-Cl cotransporter function and mRNA expression were assayed by 86Rb uptake and Northern analysis. RESULTS: Bryostatin 1 reduced Cl secretion, Na-K-Cl cotransport, and cotransporter mRNA expression. Unlike phorbol esters, these effects were largely transient. In contrast to phorbol esters, bryostatin 1 did not decrease barrier function. CONCLUSIONS: Bryostatin 1 transiently inhibits Na-K-Cl cotransport and Cl secretion, possibly through a PKC isoform also targeted by phorbol esters. Unlike phorbol esters, bryostatin 1 does not impair barrier function. The data imply that bryostatin 1 and phorbol esters differentially affect a PKC isoform involved in junctional regulation, and that epithelial transport and barrier function may be regulated by distinct PKC isoforms.     

Different behaviour of radioiodinated human recombinant interleukin-1 and its receptor antagonist in an animal model of infection

Recently, we demonstrated that radiolabelled interleukin-1alpha (IL-1) specifically accumulates in focal infection in mice through interaction with its receptor. Unfortunately, systemic side-effects of IL-1 limit its clinical application. We investigated whether this problem could be circumvented by using the interleukin-1 receptor antagonist (IL-1ra), an equally sized protein that binds to the same receptors as IL-1 without induction of biological effects. Biodistribution of 125I-IL-1 and 125I-IL-1ra was determined in Swiss mice with Staphylococcus aureus-induced abscesses in the left calf muscle at 4, 12, 24 and 48 h after injection of either 0.4 MBq 125I-IL-1 or 0.4 MBq 125I-IL-1ra. In vitro, the proteins displayed similar binding characteristics. High-performance liquid chromatographic analysis revealed a tendency for IL-1ra to associate with serum proteins. Both proteins rapidly cleared from most organs. However, the abscess uptake of 125I-IL-1ra was significantly lower than that of 125I-IL-1 at all time points (48 h p.i.: 0.06+/-0. 01%ID/g vs 0.60+/-0.04%ID/g; P<0.02). The abscess-to-contralateral muscle ratios did not exceed 15.5+/-2.9 for 125I-IL-1ra, while the ratios for 125I-IL-1 reached 46.9+/-5.7 at 48 h p.i. Despite similar in vitro receptor binding, the abscess uptake of IL-1ra was much lower than that of IL-1. The interaction of IL-1ra with serum proteins in vivo may reduce its availability for receptor binding in the infection. Although on theoretical grounds IL-1ra is very interesting, these characteristics will prevent its development as a clinically useful radiopharmaceutical to image infection.     

Breathing measurement reduces false-negative classification of tachypneic preextubation trial failures

A method using a parameter from the field of nonlinear dynamics to quantify the variability of ventricular premature complexes (VPCs) is presented. One hundred patients with coronary artery disease and > or = 10 VPCs/hour were included in the study. The RR intervals were plotted in a three-dimensional artificial phase space, and the structures in phase space were quantified by the local scaling indices, alpha. In the frequency distribution histogram, n(alpha), for each patient, the maximum of the ventricular ectopies alpha VPC, adjusted to the VPC frequency, was assessed; alpha VPC was used as the risk indicator. Endpoints were total mortality and sudden cardiac death. During follow-up (mean 3.1 years), 28 out of 100 patients died, 16 suddenly; alpha VPC had a significant prognostic impact and was independent from other risk indicators, such as left ventricular ejection fraction (LVEF). Patients who died during follow-up were characterized by a high alpha VPC. The optimal discrimination of high risk patients and low risk patients occurred at alpha VPC = 3.0. After 4 years, the survival rate of patients with a alpha VPC > 3.0 was 59%, in contrast to 97% in patients with alpha VPC < or = 0.3. As to the sudden death mortality, the survival rates were 74% and 97%, respectively. The difference between the groups were significant for both endpoints. Patients with an increased VPC variability (i.e., alpha VPC > 3.0) were at enhanced risk of sudden death and total mortality risk; alpha VPC was independent from other risk indicators such as the LVEF or heart rate variability parameters.     

Configuration of unipolar atrial electrograms during electrically induced atrial fibrillation in humans

BACKGROUND: During atrial fibrillation (AF), the atrium is activated by multiple wavelets that continuously change in size and direction. The aim of this study was to correlate the temporal variation in AF electrogram configuration with the varying spatial patterns of activation. METHODS AND RESULTS: In a group of 25 Wolff-Parkinson-White patients undergoing cardiac surgery, the free wall of the right atrium was mapped (244 points) during electrically induced AF. The unipolar electrograms recorded during 4 seconds of AF were classified into four categories: (1) single deflections, (2) short-double potentials, (3) long-double potentials, and (4) fragmented potentials. The proportion of these four types of electrograms during AF was as follows: singles, 77 +/- 12%; short-doubles, 7 +/- 3%; long-doubles, 10 +/- 7%; and fragmented, 6 +/- 4%. Electrogram morphology was an indicator for rapid uniform conduction (single potentials; positive predictive value [PPV] of 0.96), collision (short-double potentials; PPV of 0.33), conduction block (long-double potentials; PPV of 0.84), and pivoting points or slow conduction (fragmented potentials; PPV of 0.87). In type I, II, and III AF, the proportion of long-double potentials was 4 +/- 2%, 12 +/- 3%, and 18 +/- 7% (P < .05); the proportion of fragmented complexes was 2 +/- 2%, 6 +/- 3%, and 10 +/- 4% (P < .05), respectively. During electrically induced and self-terminating episodes of AF, no preferential anatomic sites for double or fragmented potentials were found in the right atrium. CONCLUSIONS: The morphology of single unipolar electrograms during AF reflects the occurrence of various specific patterns of conduction. This might be used to differentiate between different types of AF and to identify regions with structural conduction disturbances involved in perpetuation of chronic AF.     

Abdominal aortic thrombosis mimicking aortic coarctation in a newborn with heart failure]

The case of a neonate in heart failure with the classical signs of coarctation of the aorta is described. Two dimension and Doppler echocardiography ruled out coarctation of the aorta and an abdominal ultrasonography detected a large thrombotic formation in the abdominal aorta, confirmed at necropsy.