Recovery of heart tissue following focal injury induced by dietary restriction of potassium.

Sixty-four Sprague-Dawley rats (initially weighing 200-225 grams) were divided into three groups. Group 1, the experimental group, was fed a potassium depleted diet for 42 days, followed by a potassium repleted diet for up to an additional 14 days. Group 2, the dietary control group, received a potassium deficient diet, but was continuously supplemented by drinking water containing potassium chloride 150 meq/L. Group 3, the control group remained on normal rat chow and tap water during the entire investigation. Quantitative morphometric analysis was used to assess the percent of myocardium occupied by lesion. These data were analyzed by an analysis of variance (ANOVA) for repeated measures, comparing the three groups with one another; a second analysis compared the myocardial lesions of the dietary experimental group during the potassium depletion and repletion periods. At the end of the dietary depletion period (day 42) focal areas of cardiac myocyte necrosis and mononuclear infiltrate were found in the experimental group. Morphometric assessment on day 42 revealed a volume fraction (Vv) of 8.61 (+/- 4.41)%, which was significantly greater (p = 0.0018), as compared with both control groups. Lesion area significantly regressed in two and one half days after potassium was supplemented in the dietary experimental group to 0.58 (+/- 0.34)% Vv (p = 0.0005). Six days after potassium was replaced in the diet, there was no significant difference between the experimental and control groups, and only a limited connective tissue scar was noted in the experimental group. The mechanism of the rapid regression of lesions and the production of only limited connective tissue scar is suggested but requires further elucidation.

Effects of electrical stimulation on lymphatic flow and limb volume in the rat.

BACKGROUND AND PURPOSE: The mechanism by which electrical stimulation affects edema has not been elucidated. The purpose of this study was to determine whether subcontraction high-voltage stimulation (SC-HVS) (ie, electrical stimulation that did not elicit a visible contraction) applied to the right hind limbs of rats would (1) alter the rate of lymphatic uptake of injected albumin labeled with Evans blue dye (AL-EBD) and (2) affect experimentally induced edema. SUBJECTS AND METHODS: The paws of 28 anesthetized Sprague-Dawley rats (mean weight = 263 g, SD = 48 g) were injected with AL-EBD. The experimental group (n = 13) received 1 hour of SC-HVS, and the control group (n = 15) received sham treatment consisting of the same treatment administered to the experimental group but without the SC-HVS. Blood samples and volume measurements were obtained at intervals over a 7-hour period. RESULTS: Analysis of variance and post hoc testing indicated that higher amounts of AL-EBD were taken up by the lymph of the experimental group animals as compared with the control group animals at each time period following the treatment. The experimental group's AL-EBD reached significance immediately after treatment, whereas the control group required an additional 4 hours. There was no significant reduction in limb volume in either group. CONCLUSION AND DISCUSSION: The SC-HVS significantly increased the uptake of AL-EBD by lymphatic vessels, but it did not cause a significant decrease in the induced edema. The results of this study indicate that SC-HVS has the potential to reduce edema by increasing lymphatic uptake of proteins.

The role of the lymphatics in aiding regression of hypokalemic lesions in rat cardiac muscle.

This study describes the role of lymphatics in the removal of macrophages from inflammatory lesions in the heart of hypokalemic rats and rats recovering from hypokalemia. The inflammatory lesions are characterized by focal cardiomyocyte necrosis, edema, and mononuclear infiltrate. The vascular and lymphatic capillaries are maintained along with the basement membrane of the necrotic cardiomyocyte. Through prior investigation, it was revealed that refeeding potassium led to a rapid reduction in lesion area. The purpose of the current investigation was to establish the role of the lymphatics as a means of reducing the lesion area by removal of the cellular infiltrate and edema. Using a limited potassium diet, hypokalemic rats were sacrificed via perfusion fixation during the hypokalemic and the potassium re-supplementation periods. Heart tissue was examined by light and electron microscopy. During the hypokalemic period, phagocytic mononuclear cells were found engulfing necrotic cardiac muscle cells. With refeeding of potassium, these phagocytic cells appeared to be diminished in number, a reduction that coincided with a decrease in the lesion size. Lymphatic channels were dilated and full of mononuclear cells. These channels were differentiated from the vascular capillaries by standard morphological criteria. In conclusion, the lymphatics play an important role in the healing process by reducing the lesion size through the removal of phagocytic cells and the uptake of proteinaceous material.

The C-terminal domain of SIN1 in yeast interacts with a protein that binds the URS1 region of the yeast HO gene.

A protein or protein complex has previously been identified in Saccharomyces cerevisiae which both binds a short DNA sequence in URS1 of HO and interacts with SIN1. SIN1, which has some sequence similarity to mammalian HMG1, is an abundant chromatin protein in yeast and is thought to participate in the transcriptional repression of a specific family of genes. SIN1 binds DNA weakly, though it has no DNA binding specificity. Here we address the nature of the interaction between SIN1 and the specific DNA binding protein(s) to HO DNA. We show that the isolated C-terminal region of SIN1 can interact in vitro with the DNA binding protein, causing a supershift in a gel mobility shift assay. Interestingly, inclusion of the region in SIN1 which contains two acidic sequences, precludes the binding of recombinant protein to the DNA/protein complex.

Opposing effects of tyrosine kinase inhibitors on mineralization of normal and tumor bone cells.

Induction of matrix maturation and mineralization in calcified tissues is important for patients with primary bone tumors and other bone deficiencies, e.g., osteoporosis. For the former it signifies a better prognosis in osteosarcoma, and for the latter it might improve bone remodeling. In the present study we exposed osteosarcoma cells (Saos2), normal bone cells, and marrow stroma to two different tyrosine kinase (TK) inhibitors: AG-555 and AG-1478. These tyrphostins differ in their effect on signal transduction downstream to the TK receptor (RTK): AG-1478 inhibits src family TKs whereas AG-555 inhibits nuclear TKs. We found that both tyrphostins at 50 microM increased specific alkaline phosphatase (ALP) activity in Saos2 cells. AG-555 abrogated mineralization whereas AG-1478 increased it. Similarly, in human bone-derived cell cultures the same dose of tyrphostins had an opposing effect on mineralization but, in contrast to AG-555, AG-1478 positively selected cells with ALP activity. These tyrphostins also differed in their effect on rat marrow stromal cells. AG-555 decreased cell counts unselectively, whereas the decreased cell counts by AG-1478 resulted in selection of osteoprogenitor cells as indicated by a concordant increase in specific ALP activity. The effect of a lower dose of AG-1478, 5 microM, on the increase in mineralization exceeded its own efficiency in selecting cells with specific ALP activity. Our results indicate that AG-1478 selects and preserves the osteoblastic phenotype, at doses moderately higher than those required to induce mineralization, and substantially higher than the doses required for RTK inhibition. Identification of downstream molecular targets for AG-1478, in marrow stromal cells, might prove useful in designing more selective drugs, capable of separating proliferative from differentiation-inducing activities.