Synergism between nifedipine and cyclosporine A on the incorporation of [35S]sulfate into human gingival fibroblast cultures in vitro.

BACKGROUND AND OBJECTIVE: We assessed the effects of cyclosporine A and nifedipine on the in vitro incorporation of [(35)S]sulfate into gingival fibroblast cell cultures derived from responder and nonresponder subjects who had received an organ transplant followed by a therapeutic regimen using a combination of those drugs. MATERIAL AND METHODS: Gingival fibroblasts were isolated from responder and nonresponder subjects and maintained in vitro. Prior to cell harvest, gingival interleukin-1beta concentrations were determined by enzyme-linked immunosorbent assay (ELISA). Cells were untreated or exposed to either 10(-7)-10(-10) m nifedipine or 100-500 ng/ml cyclosporine A. Incorporation of [(3)H]proline or [(35)S]sulfate into the cell cultures was determined by liquid scintillation analysis. In addition, the effects of 400 ng/ml cyclosporine A + 10(-7) m nifedipine and 400 ng/ml cyclosporine A + 10(-10) m nifedipine on incorporation of [(35)S]sulfate into the cell cultures was determined. Data were compared by factorial analysis of variance (anova) and a posthoc Tukey's test. RESULTS: Gingiva from responders contained significantly more interleukin-1beta than gingiva from nonresponders (p < 0.01). The cell cultures derived from responders incorporated significantly more [(35)S]sulfate than those derived from nonresponders following exposure to either cyclosporine A or 10(-7) m nifedipine. In addition, the exposure of fibroblasts derived from gingival overgrowth to either 400 ng/ml cyclosporine A + 10(-7) m nifedipine or 400 ng/ml cyclosporine A + 10(-10) m nifedipine significantly increased or decreased, respectively, the incorporation of [(35)S]sulfate into the cultures. CONCLUSION: The therapeutic combination of cyclosporine A and nifedipine could be a significant risk factor for gingival overgrowth in subjects susceptible to either agent. The mechanism for overgrowth could include edema secondary to increased sulfated-glycosaminoglycan (sGAG) synthesis by fibroblasts, but further investigation is required.

Effects of doxorubicin on human dental pulp cells in vitro.

There is substantial information concerning the effects of continuous exposure to supratherapeutic or therapeutic concentrations of doxorubicin on human molar pulpal cells; the effects of continuous exposure to subtherapeutic concentrations of this agent are undetermined. To this end, we studied the proliferation of human fibroblasts and pulpal cells and their pattern of mineralized nodule deposition in vitro. Cell proliferation was assessed at 1, 3, 5, and 7 days from populations with either no exposure (control) or exposure to 10(-6)-10(-9) mol/L doxorubicin. Mineralized nodule deposition and calcium-45 incorporation were assessed at 7 and 21 days of culture. Data were compared by factorial ANOVA and a post-hoc Tukey test. 10(-6) and 10(-7) mol/L doxorubicin significantly reduced the total number of viable pulpal cells in cultures from days 1 to 3 (p < 0.05); doxorubicin 10(-6)-10(-9) mol/L significantly inhibited cell proliferation (p < 0.05) and DNA synthesis 5 days after plating (p < 0.001). After 21 days, doxorubicin 10(-6)-10(-8) mol/L significantly decreased calcium-45 incorporation into pulpal cultures (p < 0.001); all dilutions significantly reduced the number of mineralized nodules within the 21-day pulpal cultures (p < 0.05). In addition, all dilutions of doxorubicin significantly inhibited fibroblast cell proliferation and incorporation of [(3)H]thymidine. In contrast, the fibroblast cultures did not produce mineralized nodules, suggesting that the mineralized nodules within the pulpal cell cultures did not result from dystrophic calcification. Thus, exposure to subtheraputic doxorubicin concentrations has potential adverse effects on mineralized tissue formation within the pulp, which could affect the rates of reparative dentin deposition within the tooth pulps of patients receiving this chemotherapeutic agent.

Development and testing of the Urinary Incontinence Scales.

The Urinary Incontinence Scales were developed to measure RNs' attitude, belief, practice, and knowledge about urinary incontinence in adults. The purpose of this article is to introduce the scales and discuss their development and testing. These scales include the Urinary Incontinence Attitude Scale, the Urinary Incontinence Belief Scale, the Urinary Incontinence Practice Scale, and the Urinary Incontinence Knowledge Scale. Evaluation using confirmatory factor analysis showed the attitude, belief, and practice scales to be reliable and valid. Reliability of the knowledge scale was slightly lower than desired using both Cronbach's alpha coefficient and squared multiple correlation. While the hypothesized measurement model demonstrated acceptable goodness-of-fit, further testing is needed for generalizability of findings.

Enhancement by sodium orthovanadate of the formation and mineralization of bone nodules by chick osteoblasts in vitro.

Orthovanadate is a known inhibitor of phosphotyrosyl protein phosphatase and is reported to stimulate osteogenic cell proliferation and differentiation when administered during the logarithmic growth phase and to potentiate the mitogenic effects of several growth factors. There is little information concerning the effects of orthovanadate on bone matrix deposition and mineralization, although there is some evidence that it increases collagen synthesis by osteogenic cells. To test the effects of orthovanadate on bone nodule formation and mineralization, osteogenic cells were exposed to 5-50 microM orthovanadate or 10(-7) M insulin-like growth factor-1 for 3, 7, and 21 days after plating. Exposure to orthovanadate produced differential effects on cellular proliferation and alkaline phosphatase activity following completion of the logarithmic growth phase, and on resultant bone nodule formation and mineralization by these populations. The effects of orthovanadate on osteogenic cultures were concentration dependent: 5 microM concentrations produced by a relatively large quantity of poorly mineralized matrix, while 30-50 microM concentrations produced a smaller quantity of heavily mineralized matrix. Thus, orthovanadate could possibly be used as a growth factor for bone, if administered at the critical dosage at the proper stage of the life cycle of the osteoblast.

Site-specific variations in metabolism by human fibroblasts exposed to nifedipine in vitro.

Nifedipine induces overgrowth of gingival tissues in some patients. However, other collagenous tissues in their body do not overgrow. The purpose of this study was to compare effects of serial dilutions of nifedipine on the in vitro metabolism of fibroblasts derived from normal gingiva, nifedipine-induced hyperplastic gingiva, knee capsular ligament, and dermis. The data suggested that nifedipine affects the metabolism of fibroblasts derived not only from gingiva, but also from other collective connective tissues. Thus, nifedipine-responder cells are present in tissues other than gingiva. There was an inverse relationship between in vivo tissue levels of IL-1-beta and in vitro responsiveness to nifedipine of fibroblasts derived from that tissue. Nifedipine-induced overgrowth of connective tissues, other than gingiva, probably does not occur either because of the relatively slow rate of collagenous protein synthesis by resident fibroblasts or because of alterations in collagen deposition/resorption within susceptible tissues produced by nifedipine on collagenase synthesis.

The effects of smokeless tobacco extract on bone nodule formation and mineralization by chick osteoblasts in vitro.

Short-term exposure to smokeless tobacco extracts (STE) reportedly inhibits osteoblast metabolism. The objective of this study was to determine the effects of serial dilutions of a water-soluble extract of smokeless tobacco on osteoblast proliferation and their potential to form and mineralize bone nodules. STE significantly stimulated cell proliferation when diluted 10(2)-10(4) times; 10(3) and 10(4) dilutions produced the greatest effect. 10(2)-10(4) STE dilutions significantly increased alkaline phosphatase activity at day 7 but 10(6) STE dilutions significantly decreased it. 10(3) and 10(4) dilutions significantly increased bone nodule formation, but inhibited their mineralization. In contrast, 10(5) and 10(6) dilutions significantly decreased bone nodule formation, but increased their mineralization. Stimulation of in vitro bone nodule formation by STE was similar to that produced by 10(-7) M insulin-like growth factor 1 (IGF-1) in vivo. Heat and acid treatment of STE significantly reduced its beneficial effect on cell proliferation, suggesting that a peptide within STE may be responsible for enhancement of osteogenic cell proliferation. Thus, STE may contain a peptide capable of significantly stimulating osteoblast proliferation, differentiation and metabolism, similar to the effects of IGF-1. This peptide could have potential therapeutic benefits.

Intermittent clean self-catheterization in clients with neurogenic bladder resulting from multiple sclerosis.

For the client with multiple sclerosis (MS), urinary retention is a symptom that must be dealt with effectively because of the risk of life-threatening complications. Estimates of bladder involvement in these clients range as high as 73-90%. Intermittent clean self-catheterization (ICSC) is now being used by some clients with neurogenic bladder resulting from MS. This article addresses the success of, and problems presented by this technique to the person with MS as recorded in the literature and as related in three actual case studies.

A pubococcygeal exercise program for simple urinary stress incontinence: applicability to the female client with multiple sclerosis.

Five women with multiple sclerosis (MS) and simple urinary stress incontinence were informally evaluated and taught a program of pubococcygeal exercises to determine if inadvertent loss of urine during activities which increase intraabdominal pressure could be improved or eliminated. A correlation seemed to exist between the client's ability to experience vaginal contraction during sexual climax and elimination or improvement of simple urinary stress incontinence using a pubococcygeal exercise program. The purpose of this article is to encourage nurses to consider a pubococcygeal teaching program for treatment of simple urinary stress incontinence in female clients with MS who do experience vaginal contraction during sexual climax.

Age as a variable in an exercise program for the treatment of simple urinary stress incontinence.

This study compared the effectiveness of biofeedback on pubococcygeal muscle strengthening and simple urinary stress incontinence in older and younger women. Women aged 55 years and older and women younger than 55 years of age were taught Kegel exercises, using biofeedback, for the treatment of simple urinary stress incontinence. Eighty percent of the younger group and 67% of the older group eliminated stress incontinence. Also, the younger women required less pubococcygeal strength than the older women to control incontinence.

Rapid calcium release from cardiac sarcoplasmic reticulum vesicles is dependent on Ca2+ and is modulated by Mg2+, adenine nucleotide, and calmodulin.

A subpopulation of canine cardiac sarcoplasmic reticulum vesicles has been found to contain a "Ca2+ release channel" which mediates the release of intravesicular Ca2+ stores with rates sufficiently rapid to contribute to excitation-contraction coupling in cardiac muscle. 45Ca2+ release behavior of passively and actively loaded vesicles was determined by Millipore filtration and with the use of a rapid quench apparatus using the two Ca2+ channel inhibitors, Mg2+ and ruthenium red. At pH 7.0 and 5-20 microM external Ca2+, cardiac vesicles released half of their 45Ca2+ stores within 20 ms. Ca2+-induced Ca2+ release was inhibited by raising and lowering external Ca2+ concentration, by the addition of Mg2+, and by decreasing the pH. Calmodulin reduced the Ca2+-induced Ca2+ release rate 3-6-fold in a reaction that did not appear to involve a calmodulin-dependent protein kinase. Under various experimental conditions, ATP or the nonhydrolyzable ATP analog, adenosine 5'-(beta, gamma-methylene)triphosphate (AMP-PCP), and caffeine stimulated 45Ca2+ release 2-500-fold. Maximal release rates (t1/2 = 10 ms) were observed in media containing 10 microM Ca2+ and 5 mM AMP-PCP or 10 mM caffeine. An increased external Ca2+ concentration (greater than or equal to 1 mM) was required to optimize the 45Ca2+ efflux rate in the presence of 8 mM Mg2+ and 5 mM AMP-PCP. These results suggest that cardiac sarcoplasmic reticulum contains a ligand-gated Ca2+ channel which is activated by Ca2+, adenine nucleotide, and caffeine, and inhibited by Mg2+, H+, and calmodulin.

Single channel and 45Ca2+ flux measurements of the cardiac sarcoplasmic reticulum calcium channel.

Purified canine cardiac sarcoplasmic reticulum vesicles were passively loaded with 45CaCl2 and assayed for Ca2+ releasing activity according to a rapid quench protocol. Ca2+ release from a subpopulation of vesicles was found to be activated by micromolar Ca2+ and millimolar adenine nucleotides, and inhibited by millimolar Mg2+ and micromolar ruthenium red. 45Ca2+ release in the presence of 10 microM free Ca2+ gave a half-time for efflux of 20 ms. Addition of 5 mM ATP to 10 microM free Ca2+ increased efflux twofold (t1/2 = 10 ms). A high-conductance calcium-conducting channel was incorporated into planar lipid bilayers from the purified cardiac sarcoplasmic reticulum fractions. The channel displayed a unitary conductance of 75 +/- 3 pS in 53 mM trans Ca2+ and was selective for Ca2+ vs. Tris+ by a ratio of 8.74. The channel was dependent on cis Ca2+ for activity and was also stimulated by millimolar ATP. Micromolar ruthenium red and millimolar Mg2+ were inhibitory, and reduced open probability in single-channel recordings. These studies suggest that cardiac sarcoplasmic reticulum contains a high-conductance Ca2+ channel that releases Ca2+ with rates significant to excitation-contraction coupling.

Effects of a prenatal teaching program on postpartum regeneration of the pubococcygeal muscle.

A quasi-experimental design was used to compare postpartum perineometer readings of an experimental group of 32 women receiving a prenatal teaching program for the use of Kegel exercises with a control group of 30 women for whom the instructional program was omitted. All subjects received postpartum instruction from a pre-existing program presented by hospital staff. Significantly higher mean postpartum perineometer readings (t = 4.07; P less than .01) were found for the experimental group than for the control group, which supported the hypothesis that women who are offered the prenatal program for the use of Kegel exercises during the third trimester of pregnancy and are instructed on Kegel exercises by the staff during the hospitalized postpartum period will have significantly higher readings on the perineometer at the postpartum visit than those women for whom the prenatal program is omitted.

Interference by cortisone with endotoxin's adjuvator action on transplantation of a mouse tumour.

Observations on the in vivo plating of mouse mammary tumour are extended by making counts of tumours at a significantly earlier phase of development than in previously reported work. In the experiments now described, most of the growth of the tumours has been without benefit of stroma. The noteworthy economy of the experimental method is discussed. The persistence of endotoxin's adjuvator effect on such tumour counts is tested in the face of gamma irradiation and cortisone. Cortisone, it is found, offsets endotoxin's adjuvator action; irradiation does not. Antagonism between endotoxin and cortisone, in this system with tumour cells plated in vivo, seems to indicate that endotoxin's enhancing effect depends more on inflammatory than on immunological factors.