Paramecium calcium channels are blocked by a family of calmodulin antagonists.

Although the voltage-sensitive Ca channel present in Paramecium has been subjected to detailed physiological and genetic analysis, no organic ligands have been described that block this channel with high affinity and that ultimately can be used to identify channel components. Based on a previous observation that the naphthalene sulfonamide calmodulin antagonist W-7 can block Paramecium Ca channels at high concentration, we have synthesized analogs of W-7 that block these channels at concentrations of less than 1 microM. The effectiveness of these compounds was tested both by a sensitive behavioral assay and on Ca channels that had been incorporated into planar lipid bilayers. Despite the fact that these compounds are effective Paramecium calmodulin antagonists, two independent lines of evidence suggest that W-7 and its analogs block the Ca channel by a mechanism that is independent of their action on calmodulin. In addition, the sensitivity to W-7 or dihydropyridines of Ca channels present in a number of eukaryotic phyla has been used to identify similarities in Ca channels from widely diverse organisms. It appears that the pharmacological specificity provides a means to group Ca channels.     

Association of diurnal variations in hypothalamic but not cortical opiate ([3H]-naloxone)-binding sites with the ability of naloxone to induce LH release in the prepubertal female rat

We report the existence of a diurnal variation in the binding of the opiate antagonist [3H]-naloxone to slices of the mediobasal hypothalamus from prepubertal female rats. The binding is highest in the early morning and reaches a nadir in the late afternoon. Opiate binding in cortical slices from such animals is constant over the course of the day. Changes in receptor density, and not in receptor affinity, account for the diurnal variation in the amount of ligand bound. These diurnal variations in receptor numbers are associated with changes in the ability of naloxone to release LH and may be crucial in the transition from the juvenile state to one of competent reproductive functioning.     

Evaluation of the debonding characteristics of 2 ceramic brackets: an in vitro study.

The objectives of this study were to evaluate and compare the shear bond strengths and bond failure locations of 2 currently available orthodontic ceramic brackets. Forty polycrystalline ceramic brackets (Clarity, 3M Unitek, Monrovia, Calif) and 40 monocrystalline ceramic brackets (Inspire, Ormco, Orange, Calif) were bonded to 80 extracted premolars with the same bonding system. All bonded specimens were placed in distilled water for 42 hours at 37 degrees C followed by thermal cycling for 700 complete cycles. Forty ceramic brackets, 20 of each type, were tested on a universal testing machine to determine the shear force levels required to debond them. Forty ceramic brackets were removed with the debonding pliers recommended by the manufacturers. All teeth were examined under an optical microscope, and the adhesive remnant index was used to assess the bond failure locations. The mean shear bond strength of the Clarity brackets was 21.67 +/- 5.19 MPa, and the mean shear bond strength of the Inspire brackets was 20.32 +/- 8 MPa. The mean shear bond strengths of both brackets were higher than those considered clinically optimal. Most of the brackets (85% of Clarity and 75% of Inspire) tested on the machine failed at the bracket-adhesive interface. One premolar bonded with an Inspire bracket had enamel fracture upon debonding. Most of the brackets (90% of Clarity and 95% of Inspire) debonded with pliers failed at the bracket-adhesive interface. No enamel damage was evident in any specimen when the brackets were removed with the appropriate pliers. The results indicate that the safest way to remove ceramic brackets with respect to reducing the chance of enamel damage is to use the debonding technique specifically designed for each.     

Quality of life and exercise performance in patients in sinus rhythm versus persistent atrial fibrillation: a Veterans Affairs Cooperative Studies Program Substudy.

OBJECTIVES: The purpose of this study was to determine quality of life (QOL) and exercise performance (EP) in patients with persistent atrial fibrillation (AF) converted to sinus rhythm (SR) compared with those remaining in or reverting to AF. BACKGROUND: Restoration of SR in patients with AF improving QOL and EP remains controversial. METHODS: Patients with persistent AF were randomized double-blind to amiodarone, sotalol, or placebo. Those not achieving SR at day 28 were cardioverted and classified into SR or AF groups at 8 weeks (n = 624) and 1 year (n = 556). The QOL (SF-36), symptom checklist (SCL), specific activity scale (SAS), AF severity scale (AFSS), and EP were assessed. RESULTS: Favorable changes were seen in SR patients at 8 weeks in physical functioning (p < 0.001), physical role limitations (p = 0.03), general health (p = 0.002), and vitality (p < 0.001), and at 1 year in general health (p = 0.007) and social functioning (p = 0.02). Changes in the scores for SCL severity (p = 0.01), functional capacity (p = 0.003), and AFSS symptom burden (p < 0.001) at 8 weeks and in SCL severity (p < 0.01) and AF symptom burden (p < 0.001) at 1 year showed significant improvements in SR versus AF. Symptomatic patients were more likely to have improvement. The EP in SR versus AF was greater from baseline to 8 weeks (p = 0.01) and to 1 year (p = 0.02). The EP correlated with physical functioning and functional capacity except in the AF group at 1 year. CONCLUSIONS: In patients with persistent AF, restoration and maintenance of SR was associated with improvements in QOL measures and EP. There was a strong correlation between QOL measures and EP.     

Monocyte adhesion molecule expression in interstitial inflammation in patients with renal failure.

BACKGROUND: Patients with renal failure have an increased susceptibility to infections. We therefore studied the recruitment of monocytes and their expression of adhesion molecules CD11b and CD62L at the site of interstitial inflammation in patients with renal failure. Furthermore, we studied if the capacity of monocytes to up-regulate CD11b in interstitial inflammation was determined by the interstitial concentration of chemotactic factors. METHODS: Three intensities of interstitial inflammation (0, intermediate and intense) were established in skin blister chambers. Leukocyte count, CD11b/CD62L expression, monocyte chemotactic protein-1 (MCP-1) and blister activity in terms of CD11b mobilization were determined. RESULTS: The CD62L expression on monocytes was lower in the peripheral circulation in patients with renal failure compared with healthy subjects (P<0.005 and P<0.001). At the site of interstitial inflammation patients had a higher expression of CD62L (intermediate, P<0.05; intense, P<0.005). Furthermore, monocytes from patients had an impaired capacity to mobilize CD11b both in the peripheral circulation (P<0.005) and at the intermediate and intense sites of interstitial inflammation (P<0.005 and P<0.001, respectively) compared with cells collected from healthy subjects. We incubated monocytes in blister exudates, in order to explore whether this phenomenon is caused by cellular factors and/or to the interstitial concentration of chemotactic mediators. The expression of CD11b on monocytes from healthy blood donors incubated in blister exudates from either patients or healthy subjects in vitro was similar. The interstitial concentration of MCP-1 at the site of intermediate inflammation was significantly lower in patients with renal failure compared with the corresponding blister exudate collected from healthy subjects (P<0.05), but no differences were observed at the site of intense inflammation. Furthermore, neutralizing the action of MCP-1 in blister exudates with monoclonal antibodies did not have any impact on monocyte CD11b expression following incubation in blister exudates. CONCLUSION: These studies indicate that the impaired capacity of monocytes to mobilize CD11b at the site of inflammation in patients with renal failure is more dependent on constitutive cellular factors than the concentration of CD11b mobilizing factors in the interstitium.     

Nonheme iron in sickle erythrocyte membranes: association with phospholipids and potential role in lipid peroxidation

Previous studies documented the abnormal association of heme and heme proteins with the sickle RBC membrane. We have now examined RBC ghosts and inside-out membranes (IOM) for the presence of nonheme iron as detected by its formation of a colored complex with ferrozine. Sickle ghosts have 33.8 +/- 18.2 nmol nonheme iron/mg membrane protein, and sickle IOM have 4.3 +/- 3.0 nmol/mg. In contrast, normal RBC ghosts and IOM have no detectable nonheme iron. The combination of heme and nonheme iron in sickle IOM averages nine times the amount of membrane- associated iron in normal IOM. Kinetics of the ferrozine reaction show that some of this nonheme iron on IOM reacts slowly and is probably in the form of ferritin, but most (72% +/- 18%) reacts rapidly and is in the form of some other biologic chelate. The latter iron compartment is removed by deferoxamine and by treatment of IOM with phospholipase D, which suggests that it represents an abnormal association of iron with polar head groups of aminophospholipids. The biologic feasibility of such a chelate was demonstrated by using an admixture of iron with model liposomes. Even in the presence of tenfold excess adenosine diphosphate, iron partitions readily into phosphatidylserine liposomes; there is no detectable association with phosphatidylcholine liposomes. To examine the bioavailability of membrane iron, we admixed membranes and t-butylhydroperoxide and found that sickle membranes show a tenfold greater peroxidation response than do normal membranes. This is not due simply to a deficiency of vitamin E, and this is profoundly inhibited by deferoxamine. Thus, while thiol oxidation in sickle membranes previously was shown to correlate with heme iron, the present data suggest that lipid peroxidation is related to nonheme iron. In control studies, we did not find this pathologic association of nonferritin, nonheme iron with IOM prepared from sickle trait, high-reticulocyte, postsplenectomy, or iron-overloaded individuals. These data provide additional support for the concept that iron decompartmentalization is a characteristic of sickle RBCs.     

The introduction of low-osmolar contrast agents in radiology. Medical, economic, legal, and public policy issues

This case study of the public policy implications of introducing a new technology in radiology, namely, low-osmolar contrast media (LOCM), raises the issues of whether and how to place appropriate limits on new technologies. Although these contrast media represent small episodic costs, they may add up to an aggregate expenditure of nearly $1 billion per year if used for all contrast injections. As a result, this technology raises a number of important medical, economic, legal, and public policy questions. Our cost-effectiveness analysis and an analysis of the medical evidence suggest that LOCM should be limited to high-risk patients. We discuss in this article how the legal system might respond to such limitations, and we consider various public policy options for adopting restrictions on use. We conclude that the medical profession should take the lead in developing protocols for appropriate assessment, reimbursement, and use of LOCM.