Two functionally different Na/K pumps in cardiac ventricular myocytes

The whole-cell patch-clamp technique was used to voltage clamp acutely isolated myocytes at -60 mV and study effects of ionic environment on Na/K pump activity. In quiescent guinea pig myocytes, normal intracellular Na+ is approximately 6 mM, which gives a total pump current of 0.25 +/- 0.09 pA/pF, and an inward background sodium current of 0.75 +/- 0.26 pA/pF. The average capacitance of a cell is 189 +/- 61 pF. Our main conclusion is the total Na/K pump current comprises currents from two different types of pumps, whose functional responses to the extracellular environment are different. Pump current was reversibly blocked with two affinities by extracellular dihydro-ouabain (DHO). We determined dissociation constants of 72 microM for low affinity (type-1) pumps and 0.75 microM for high affinity (type-h) pumps. These dissociation constants did not detectably change with two intracellular Na+ concentrations, one saturating and one near half-saturating, and with two extracellular K+ concentrations of 4.6 and 1.0 mM. Ion effects on type-h pumps were therefore measured using 5 microM DHO and on total pump current using 1 mM DHO. Extracellular K+ half-maximally activated the type-h pumps at 0.4 mM and the type-1 at 3.7 mM. Extracellular H+ blocked the type-1 pumps with half-maximal blockade at a pH of 7.71 whereas the type-h pumps were insensitive to extracellular pH. Both types of pumps responded similarly to changes in intracellular-Na+, with 9.6 mM causing half-maximal activation. Neither changes in intracellular pH between 6.0 and 7.2, nor concentrations of intracellular K+ of 140 mM or below, had any effect on either type of pump. The lack of any effect of intracellular K+ suggests the dissociation constants are in the molar range so this step in the pump cycle is not rate limiting under normal physiological conditions. Changes in intracellular-Na+ did not affect the half-maximal activation by extracellular K+, and vice versa. We found DHO-blockade of Na/K pump current in canine ventricular myocytes also occurred with two affinities, which are very similar to those from guinea pig myocytes or rat ventricular myocytes. In contrast, isolated canine Purkinje myocytes have predominantly the type-h pumps, insofar as DHO-blockade and extracellular K+ activation are much closer to our type-h results than type-1. These observations suggest for mammalian ventricular myocytes: (a) the presence of two types of Na/K pumps may be a general property. (b) Normal physiological variations in extracellular pH and K+ are important determinants of Na/K pump current. (c) Normal physiological variations in the intracellular environment affect Na/K pump current primarily via the Na+ concentration. Lastly, Na/K pump current appears to be specifically tailored for a tissue by expression of a mix of functionally different types of pumps.     

State-coordinated services for traumatic brain injury survivors: toward a model delivery system

Since 1985, the state of Minnesota, through a variety of advocacy, legislative, and interagency efforts, has made incremental gains in public policy and service development for persons with traumatic brain injury (TBI). This article reviews the roles and functions of select state programs and departments in coordinating TBI services. Key initiatives, as well as the current model of public policy and services, are outlined. Current and future service development and initiatives are discussed. Finally, specific implementation recommendations are proposed.     

Elevated inosine monophosphate levels in resting muscle of patients with stable chronic obstructive pulmonary disease

From January 1981 through October 1995, a total of 28 patients underwent surgery for metastatic lung tumors in our institute. A retrospective review and survival analysis of these patients are reported. There were 12 males and 16 females, with the mean age of 57 years. Of them, 20 patients had solitary lesions. Pulmonary metastasis was from the colon in 8 patients, from the breast in 5, from the stomach and the uterus in each 3, from the bladder, the rectum and the soft tissue in each 2, and from the kidney, the ovarium and the thyroid in each 1. A lobectomy was performed in 22 patients, 17 of whom were accompanied with mediastinal lymph node dissection (R2a). Partial resection without lymph node dissection was performed in 6 patients (R0). Overall 5-year survival was 24.7%. Four-year or over survivor were 4 patients who underwent a lobectomy with R2a. The patients undergoing lobectomy had a significantly better prognosis compared with patients with partial resection (p<0. 05). The patients with R2a had significantly less local recurrence than patients with R1 or R0 (p<0.05). We conclude that lobectomy with R2a is suitable treatment for metastatic lung cancer.     

Failure to induce organ-specific autoimmunity by breaking of tolerance: importance of the microenvironment

Peripheral tolerance is considered to be a safeguard against autoimmunity. Using a TCR-transgenic mouse system displaying peripheral tolerance against a liver-specific MHC class I Kb antigen, we investigated whether the breaking of tolerance would result in autoimmunity. Reversal of tolerance was achieved by simultaneous challenge with cells expressing the Kb autoantigen and IL-2. Tolerance could not be broken with IL-2 alone or when Kb- and IL-2-expressing cells were applied to different sites of the mice. However, despite the presence of activated autoreactive T cells that were able to reject Kb-positive grafts no autoaggression against the Kb-positive liver was observed. These results indicate that breaking of tolerance per se is not sufficient to cause liver-specific autoimmunity. However, when in addition to breaking tolerance the mice were infected with a liver-specific pathogen, autoaggression occurred. Thus, in this system at least two independent steps seem to be required for organ-specific autoimmunity: reversal of peripheral tolerance resulting in functional activation of autoreactive T cells and conditioning of the liver microenvironment which enables the activated T cells to cause tissue damage.