Transcription factor phosphorylation by pp90(rsk2). Identification of Fos kinase and NGFI-B kinase I as pp90(rsk2).

The in vitro phosphorylation of transcription factors by growth factor-activated protein kinases has resulted in the discovery of a number of activities whose identities and relationships to one another are unclear. Fos kinase is a growth factor-stimulated serine/threonine protein kinase that phosphorylates c-Fos at serine 362 within the carboxyl-terminal regulatory domain. Fos kinase activation is dependent on p21(ras) and mitogen-activated protein kinase/ERK kinase kinase (MEK) activity and is independent of phosphatidylinositol 3-kinase activity. We have purified Fos kinase by affinity chromatography using the Sepharose-linked protein kinase inhibitor, bisindolylmaleimide (BIM). Fos kinase has an apparent molecular mass of 88 kDa, and mass spectrophotometric analysis of the isolated protein showed that it produced tryptic fragments identical to those predicted for pp90(rsk2). Fos kinase isolated from nerve growth factor-stimulated PC12 cells is indistinguishable from NGFI-B kinase I, based on their chromatographic behavior, substrate specificities, and relative sensitivity to BIM. Furthermore, we have distinguished Fos kinase from calcium/cAMP response element-binding protein (CREB) kinase. Therefore, Fos kinase and NGFI-B kinase I and pp90(rsk2) represent the same protein kinase species. Moreover, we report that pp90(rsk2) exists within nerve growth factor-stimulated PC12 cells as two chromatographically and immunologically distinct species. Finally, we demonstrate that CREB kinase is distinct from pp90(rsk2).

The role of cell-associated interleukin-1 in bleomycin-induced pulmonary fibrosis.

The activities of cell-associated IL-1 (IL-1 alpha) and extracellular IL-1 (IL-1 beta) in alveolar macrophages (AM) from rats with bleomycin-induced pulmonary fibrolosis were measured to determine their role in fibroblast growth. AM were obtained on Days 1, 3, 6, 9 and 12 after a intratracheal injection of bleomycin, and the IL-1 activity and fibroblast growth-stimulating activity in fixed AM and AM supernatants were measured. Higher cell-associated IL-1 activity was detected in AM from bleomycin-treated rats than in those of control on Day 1 through 9. But extracellular IL-1 activity in the supernatant of AM from bleomycin-treated rats significantly higher only on Day 1. Expression of IL-1 alpha mRNA in AM from bleomycin-treated rats was significantly higher than that in AM of control, but there was no significant difference in the mRNA levels of IL-1 beta in AM of these two groups. Fixed AM from bleomycin-treated rats caused growth-inhibition of fibroblasts in a density-dependent manner. The inhibitory activity was decreased by pretreatment of fixed AM with anti IL-1 alpha antibody, but not anti IL-1 beta antibody. These results suggest that cell-associated IL-1 (IL-1 alpha) is produced continuously in AM from rats with bleomycin-induced pulmonary fibrosis and may be important in regulation of this disorder by inhibiting fibroblast growth.

Coronary vascularization during development in the rat and its relationship to basic fibroblast growth factor.

OBJECTIVE: Our overall aims were to elucidate the temporal and spatial sequence of coronary vascularization during development in the rat, and to determine whether basic fibroblast growth factor expression corresponds to any phase of the vascularization process. METHODS: Immunohistochemical, histochemical, morphometric and in situ hybridization analyses were performed on prenatal and postnatal hearts of various ages. RESULTS: Coronary vascularization, which begins at embryonic day 13 (E13) with blood island-like structures in the epicardium, progresses from this layer toward the endocardium as indicated by a transmural gradient of vascular volume throughout the ventricles. Vascular smooth muscle first appears in E17 hearts at the time a capillary-like plexus coalesces and penetrates the aorta to form the main coronary arteries. These vessels maintain an anastomatic morphology and must undergo subsequent remodeling in order to assume adult branching characteristics. The early postnatal period is characterized by development of the arterial tree and the enzymatic differentiation of the arteriolar and venular ends of the capillary bed. Although bFGF is expressed both prenatally and postnatally, the highest mRNA expression was noted during the early period of vascularization (E14 and E15), and the early neonatal period (1-6 days) which corresponds to a period of substantial microvascular growth. CONCLUSIONS: Coronary vascularization follows a temporal sequence which includes transmural expansion of the capillary bed, arteriolar formation subsequent to vascular penetration of the aorta, and postnatal growth, differentiation, and remodeling. Since high levels of bFGF expression are correlated with key time points in coronary vascular growth, bFGF may play an important role in this process.

The effect of single bolus dose of esmolol for controlling the tachycardia and hypertension during laryngoscopy and tracheal intubation.

Tachycardia and hypertension usually accompany laryngoscopy and tracheal intubation. This response is undesirable, especially in patients with cardiovascular or intracranial diseases. Esmolol is a cardioselective, ultrashort-acting beta adrenergic blocking agent with a very short half-life. The efficacy of bolus dose of esmolol in blunting hemodynamic responses during laryngoscopy and tracheal intubation was evaluated. 45 patients (15 in each group) of ASA physical status I and II scheduled for elective non-cardiac surgery were included in this randomized, placebo-controlled study. At time zero, the study preparation (placebo, 100 or 200 mg of esmolol) was administered intravenously, followed by thiopentone 5 mg/kg and succinylcholine 1.5 mg/kg for induction. Tracheal intubation was performed 2 minutes after time zero. Anesthesia was maintained with 50% nitrous oxide and 1.0 MAC halothane in oxygen, and vecuronium 0.08 mg/kg. Heart rate (HR) and systolic blood pressure (SBP) were recorded every minute for 10 minutes. To compare with the placebo group, there was a significant decrease in either HR or SBP in 200 mg group in the 8 minutes course after intubation (p < 0.05). There was a significant decrease in HR in the 100 mg group at the 3rd, 4th, and 5th minutes when compared with the placebo group (p < 0.05). The differences in SBP between the 100 mg group and placebo group were significant at the 3rd and 4th minutes (p < 0.05). Both bolus dosages of esmolol could effectively attenuate the tachycardia and hypertension produced by laryngoscopy and tracheal intubation. Furthermore, esmolol 200 mg presented a better hemodynamic stability than esmolol 100 mg during induction of anesthesia.

Sacroepidural analgesia for post-operative pain relief in children.

For evaluation of the practicality of epidural analgesia for alleviation of post-operative pain, 66 class I or II patients with age ranging from 6 months to 12 years undergoing elective surgical procedures below the lower abdomen were enrolled for study during the period from January to April 1989. Before termination of general anesthesia a single dose of 0.25% bupivacaine, respectively at 0.25 mL/kg (Ideal body weight), 0.5 mL/kg, 0.75 mL/kg, 1.0 mL/kg and 1.25 mL/kg was given sacroepidurally to 5 groups of patients. Our results showed that for surgery below the lower abdomen a dose at 1.0 mL/kg was sufficient to suffice the need for relief of post-operative pain. At this dosage it achieved a neural block up to T8-T6 and provided with an analgesia that could last 6.0 +/- 2.1 hours. During the entire course there were no untoward effects such as hypotension, bradycardia, vomiting and shivering to come about. Therefore, sacroepidural analgesia with 0.25% bupivacaine at fitting single dose is safe and feasible in children as far as relief of post-operative pain for procedures below the lower abdomen is concerned.