Adrenergic regulation of aqueous outflow

Epinephrine increased outflow facility and cyclic AMP in the in vitro perfused human anterior segment with a maximal facility increase of 44% occurring at approximately 2 x 10(-5) M. Cyclic AMP measured in the perfusate from anterior segments increased by 12-14 fold after administration of 10(-5) M epinephrine. Both the facility increase and cyclic AMP rise were blocked by the beta-2 selective antagonist, ICI118,551. While there was a correlation between the facility increase and elevation in cyclic AMP levels, the rise in cyclic AMP preceded the facility increase by about 1 hour, suggesting that the ultimate effect of epinephrine involved a rather slow event such as synthesis and release of prostaglandins or protein synthesis. Subsequent perfusion studies showed that very large concentrations of indomethacin were necessary to block the outflow facility effect of epinephrine, suggesting that prostaglandin synthesis did not underlie the facility effect in this system. However, 5 x 10(-5) M cyclohexamide blocked the effect on outflow facility of both epinephrine and forskolin, but did not block the rise in cyclic AMP. These studies suggest that protein synthesis may play a role in the epinephrine-induced facility increase at some point beyond the second messenger level.     

A cytosolic granzyme B inhibitor related to the viral apoptotic regulator cytokine response modifier A is present in cytotoxic lymphocytes

Using a polymerase chain reaction strategy we identified a serine proteinase inhibitor (serpin) in human bone marrow that is related to the cellular serpin proteinase inhibitor 6 (PI-6) and the viral serpin cytokine response modifier A (CrmA). This serpin, proteinase inhibitor 9 (PI-9), has an unusual reactive center P1(Glu)-P1'(Cys), which suggests that it inhibits serine proteinases that cleave after acidic residues. The only known serine proteinase with this specificity is granzyme B, a granule cytotoxin produced by cytotoxic lymphocytes. To test the interaction of PI-9 with granzyme B we prepared recombinant hexa-histidine tagged PI-9 in a yeast expression system. Addition of the recombinant protein to native granzyme B resulted in an SDS-resistant complex typical of serpin-serine proteinase interactions. Further analysis showed that complex formation followed bimolecular kinetics with a second order rate constant of 1.7 +/- 0.3 x 10(6) M-1 s-1, which is in the range for a physiologically significant serpin-proteinase interaction. Recombinant PI-9 also completely abrogated granzyme B and perforin-mediated cytotoxicity in vitro. Examination of PI-9 mRNA distribution demonstrated that it is expressed in immune tissue, primarily in lymphocytes. The highest levels of PI-9 mRNA and protein were observed in natural killer cell leukemia cell lines and in interleukin-2 stimulated peripheral blood mononuclear cells, which also produce granzyme B. Like PI-6, PI-9 was shown to be a cytosolic protein that is not secreted. Fractionation of natural killer cells and stimulated peripheral blood mononuclear cells demonstrated that PI-9 is in a separate subcellular compartment to granzyme B. These results suggest that PI-9 serves to inactivate misdirected granzyme B following cytotoxic cell degranulation. This may explain why cytotoxic cells are not damaged by their own granzyme B during destruction of abnormal cells.     

Cognitive distortions and adolescent affective disorder. Validity of the CNCEQ in an inpatient sample. Children's Negative Cognitive Error Questionnaire

Despite a proliferation of recent research examining childhood and adolescent depression, the area still lags behind the adult depression field, particularly in the investigation of cognitive correlates of affective psychopathology. To advance cognitive research with youth, the Children's Negative Cognitive Error Questionnaire (CNCEQ) was developed to provide a measure of cognitive distortions or errors in children and adolescents. Yet, few studies have employed the CNCEQ and no evidence exists supporting the validity of its four component cognitive error scales. The purpose of the present study was to examine the construct validity of the CNCEQ and its constituent scales through the use of factor analysis and criterion-group comparisons. Groups of adolescent psychiatric inpatients, diagnosed as affective or disruptive disordered, completed the CNCEQ following admission. Results failed to support the implicit four-factor structure of the CNCEQ, instead suggesting the appropriateness of a single-factor solution labeled "negative thinking." Despite no diagnostic group differences on the CNCEQ total or other scale scores, affective disordered patients evinced more cognitive errors on the Overgeneralizing scale. Findings suggest that the CNCEQ in its current stage of development holds promise, yet requires refinement to produce a valid measure of cognitive functioning in youth.     

Preliminary investigation of the efficacy of sublingual verapamil in the management of acute atrial fibrillation and flutter

The antiarrhythmic properties of sublingual verapamil were investigated in seven patients with acute fast atrial flutter (n = 2) or fibrillation (n = 5). A rapid and significant (P < 0.05) reduction in the ventricular rate was achieved in all seven patients. The ventricular rate at peak plasma verapamil concentration (+/- s.d.) was significantly slower than on admission (101.6 +/- 11.3 and 159 +/- 5.3 beats min-1 respectively, P < 0.01). The ventricular rate remained controlled for over 4 h. Sublingual verapamil was rapidly absorbed with the maximum peak plasma concentration (153.3 +/- 15.5 ng ml-1) being achieved after 1.21 +/- 0.18 h. Side-effects of sublingual verapamil were limited to one report of a bitter taste. The sublingual administration of verapamil may provide an alternative method for the control of acute fast atrial fibrillation and flutter in selected patients.     

MR angiography in the preoperative evaluation of abdominal aortic aneurysms: a preliminary study

PURPOSE: The ability of magnetic resonance (MR) angiography to depict visceral and renal vessels was evaluated in patients with abdominal aortic aneurysms (AAAs). PATIENTS AND METHODS: MR sequences (sagittal T1-weighted, two-dimensional coronal, and three-dimensional axial time-of-flight) were compared in a prospective blinded fashion with conventional angiograms obtained preoperatively in 23 patients with AAAs. Results were correlated with surgical findings when available. RESULTS: Operative aortic clamp site was correctly predicted with conventional angiography in 95% of patients and with MR angiography in 86% (P > .1). Aneurysm neck measurements obtained with the two modalities were within 1 cm in 91% of cases. With conventional angiography as the standard of reference, 96% of all renal arteries were identified on MR angiograms but 36% of accessory arteries were missed. MR angiography enabled identification of patients who had at least one renal artery stenosis greater than 50% with a sensitivity of 100% and specificity of 89%. For identifying individual renal artery, celiac artery, and superior mesenteric artery stenoses of similar severity, the sensitivity and specificity were 67% and in excess of 96%, respectively. The celiac artery could not be evaluated in one case. CONCLUSION: The results of this small study suggest that the role of MR angiography in the preoperative evaluation of AAA warrants further investigation.     

Coordinate and noncoordinate colony stimulating factor formation by human monocytes

The regulation of macrophage colony-stimulating factor (M-CSF) formation by elutriation-purified human monocytes was studied in vitro and compared with that for granulocyte-macrophage CSF (GM-CSF) and granulocyte CSF (G-CSF). The levels of the three CSFs were measured by immunoassay. Lipopolysaccharide (LPS, 100 ng/ml) was able to enhance CSF formation but the levels were significantly influenced by the presence of a cyclooxygenase product(s) in the cultures. In the presence of LPS, both M-CSF and GM-CSF were up-regulated by cyclooxygenase inhibition (indomethacin, 10(-5) M), while G-CSF was down-regulated. Exogenous prostaglandin E2 (PGE2, 10(-7) M) reversed the actions of indomethacin. In LPS-treated cells, in contrast to the different regulation by endogenous eicosanoid of M-CSF and GM-CSF formation and G-CSF formation, both interleukin-4 (IL-4, 250 pM) and the glucocorticoid dexamethasone (10(-7) M) lowered the amounts of all CSFs. When the actions of CSFs were examined for their effects on CSF formation, M-CSF could not stimulate either GM-CSF or G-CSF synthesis, in contrast to literature findings, while GM-CSF enhanced M-CSF formation but not that of G-CSF. These studies indicate that there can be both coordinate and noncoordinate control of CSF expression by human monocytes.     

Single myosin molecule mechanics: piconewton forces and nanometre steps

A new in vitro assay using a feedback enhanced laser trap system allows direct measurement of force and displacement that results from the interaction of a single myosin molecule with a single suspended actin filament. Discrete stepwise movements averaging 11 nm were seen under conditions of low load, and single force transients averaging 3-4 pN were measured under isometric conditions. The magnitudes of the single forces and displacements are consistent with predictions of the conventional swinging-crossbridge model of muscle contraction.