Hunter, Larry W.

In postmenopausal women, decreases in estrogen disrupt the normal balance of bone turnover, increasing bone resorption relative to bone formation. This study investigated the effects of menopausal hormone treatment (MHT) on volumetric bone mineral density (vBMD) and markers of bone turnover in plasma of healthy, recently‐menopausal women. Women (age 52.3±2.3 yrs; n=115) enrolled in the Kronos Early Estrogen Prevention Study (KEEPS) were randomized to either: oral conjugated equine estrogen (oCEE), transdermal 17β estradiol (tE2) each with intermittent oral micronized progesterone or placebo (Pl), for 48 months. vBMD was assessed by quantitative computed tomography of the wrist, and plasma levels of osteocalcin (OCN; a bone turnover marker), and of Fetuin‐A, a hepatocyte‐derived modulator of osteogenesis, were quantified by ELISA. At 48 months, cortical vBMD at the distal radius had decreased by 2.9% in the Pl group, correspondent with an increase above initial plasma OCN (15.0 ng/ml) of 38.5%. MHT attenuated the vBMD decrease (0.9% in both treatment groups combined; P=0.015), and the OCN increase (8.8 and 4.3% in oCEE and tE2, respectively). Plasma Fetuin‐A levels did not differ significantly among groups at 48 mo. Unexpectedly, Fetuin‐A was negatively associated (r=‐0.50 to ‐0.62; P <0.05) with changes in trabecular bone microarchitectural parameters at the distal radius in the Pl group, whereas both oCEE and tE2 attenuated this effect (all P‐values >0.05). As expected, MHT attenuated bone turnover in postmenopausal women. However, the relationship between processes regulated by OCN and Fetuin‐A require further evaluation. Grant Funding Source : Supported by Aurora Foundation, HL90639 and AG44170

Experiments were designed to investigate isolatable factors that could either promote or inhibit formation of hydroxyapatite (HA) crystals associated with mineralization of the fibrous capsule surrounding silicone breast implants. Explanted capsular tissue was minced, filtered (0.2μm), inoculated into culture plates containing DMEM/10% FBS and kept at 37°C, 95%O 2 . Within 14 days, biofilm‐like and suspending (planktonic) aggregates containing HA formed to a greater extent in plates inoculated with sample from calcified (CC) than non‐calcified (NC) capsules (158 vs 4 nephelometric turbidity units, respectively). Confocal microscopy of formalin‐fixed sections of NC showed the major cell types to be macrophages and fibroblasts. These cells were immunopositive for fetuin, a blood protein. In contrast, CC were largely fibrous; fetuin labeling was sparse except for the calcific plaque itself on the surface of the capsule adjacent to the implant. Thus, a factor which affects HA formation, possibly a HA nucleator, or a modifier of nucleator activity, can be isolated from mineralized fibrous breast capsules. Localization of fetuin within NC but not CC cells is consistent with its known role in inhibiting HA crystal growth. Supported by grants from the Fetzer Foundation, NIH HL88988 and Division of Experimental Surgery, Mayo Clinic.

Neuropeptide Y-like immunoreactivity (NPY-LI), norepinephrine (NE), and 3,4-dihydroxyphenylglycol (DOPEG), the intraneuronal metabolite of NE, were measured in superfusate before, during, and after electrical stimulation (ES) of nerves and in the tissue extract of dog pulmonary artery after in vitro superfusion. Vessels were stimulated at 12, 6, or 1 Hz. NE and DOPEG were quantified by high-performance liquid chromatography with electrochemical detection, NPY-LI by radioimmunoassay. The Krebs-Ringer superfusate was maintained at a partial pressure of oxygen of either 472, 100, 50, or 20 mmHg. Mean DOPEG efflux was 0.17, 0.15, 0.13, and 0.07 pmol/min during basal conditions and 0.73, 0.39, 0.30, and 0.15 pmol/min, respectively, during 12-Hz continuous stimulation using these four oxygen pressures. Stimulation-evoked efflux of NPY-LI was 1.5, 1.1, 0.3, and 0.2 fmol/min during 12-Hz stimulation. These studies provide evidence that the production and subsequent efflux of DOPEG into superfusate under resting conditions, during ES, and following ES are oxygen sensitive. Additionally, the efflux of NPY-LI from dog pulmonary artery resulting from high frequencies of ES is oxygen sensitive.

Dihydroxyphenylglycol (DOPEG), the metabolite of norepinephrine (NE) that arises intraneuronally, was measured together with NE in superfusates collected before, during, and after nerve stimulation and in extracts of dog saphenous vein after superfusion and electrical stimulation (ES). Different concentrations of NE in the synaptic clefts were achieved by treating tissues with corticosterone, corticosterone and yohimbine, corticosterone and cocaine, or by omitting drugs from the superfusate. NE and DOPEG were quantitated by liquid chromatography with electrochemical detection. The time courses of NE overflow and DOPEG efflux into superfusate were followed. The amounts of DOPEG in superfusates under basal conditions were two to four times higher than the amounts of NE and progressively increased during ES except in tissues with neuronal uptake inhibited. NE overflow reached a steady state within the first 6 min of ES. Increased NE concentrations in synaptic clefts resulted in increased DOPEG production except where neuronal uptake was inhibited. The increased DOPEG production during ES appears to reflect the increased rate of neuronal uptake, which results in more NE being available for intraneuronal metabolism. No evidence was found that newly formed DOPEG was delayed in leaving the tissue. Thus the increase in DOPEG production that occurs during ES may be useful as an index of neuronal uptake of NE in dog saphenous vein.

Previously, the biological factors were weighed by the expert opinions. In this paper, we evaluate the risk factors proposed by the expert opinions. We use fuzzy logic techniques for predicting risk for development of carotid intima–medial thickness (CIMT) using expert opinion and various factors associated with CIMT in recently menopausal women. Study participants were enrolled in the Kronos Early Estrogen Prevention Study (KEEPS), a four-year study where participants were assigned to either oral or transdermal hormone treatments or to placebo. We use preference modeling techniques to determine the consensus winner of the causal factors. We use a measure to determine the degree to which one factor is preferred to another. We also determine the extent to which a factor is preferred to another by "most" experts. We determine the degree to which the experts agree. We consider a model concerning social networks and the effects of these networks on persons' opinions.

We hypothesized that 1) hypothyroidism (Hyp) decreases myosin heavy chain (MHC) content per half-sarcomere in diaphragm muscle (Diam) fibers, 2) Hyp decreases the maximum specific force (Fmax) of Diamfibers because of the reduction in MHC content per half-sarcomere, and 3) Hyp affects MHC content per half-sarcomere and Fmaxto a greater extent in fibers expressing MHC type 2X (MHC2X) and/or MHC type 2B (MHC2B). Studies were performed on single Triton X-permeabilized fibers activated at pCa 4.0. MHC content per half-sarcomere was determined by densitometric analysis of SDS-polyacrylamide gels and comparison with a standard curve of known MHC concentrations. After 3 wk of Hyp, MHC content per half-sarcomere was reduced in fibers expressing MHC2Xand/or MHC2B. On the basis of electron-microscopic analysis, this reduction in MHC content was also reflected by a decrease in myofibrillar volume density and thick filament density. Hyp decreased Fmaxacross all MHC isoforms; however, the greatest decrease occurred in fibers expressing fast MHC isoforms (∼40 vs. ∼20% for fibers expressing slow MHC isoforms). When normalized for MHC content per half-sarcomere, force generated by Hyp fibers expressing MHC2Awas reduced compared with control fibers, whereas force per half-sarcomere MHC content was higher for fibers expressing MHC2Xand/or MHC2Bin the Hyp Diamthan for controls. These results indicate that the effect of Hyp is more pronounced on fibers expressing MHC2Xand/or MHC2Band that the reduction of Fmaxwith Hyp may be at least partially attributed to a decrease in MHC content per half-sarcomere but not to changes in force per cross bridge.

Carotid intima-medial thickness (CIMT) allows for early detection of carotid atherosclerotic vascular disease. Using fuzzy logic techniques, we propose a model for calculating the risk for predicting CIMT using expert opinion and various factors associated with CIMT in recently menopausal women. Study participants were enrolled in the Kronos Early Estrogen Prevention Study (KEEPS), a four year study where participants were assigned to either oral or transdermal hormone therapy or to the placebo group. The fuzzy mathematical model uses five fuzzy logic methods to calculate this risk factor for CIMT, including the analytic hierarchy process (AHP) method, Guiasu method, Yen method, Dempster–Shafer (DS) method, and the set valued statistical method (SVSM). The four methods, AHP, Guiasu, Yen, DS, and SVSM, were correlated to each other according to the Pearson ranking method. In addition, results showed that the estrogen treatment groups showed a statically significant decrease in calculated CIMT risk compared to placebo when all the methods were combined together (p = 0.0017). Our fuzzy mathematical model provides the first fuzzy mathematical model to predict the risk of CIMT in recently menopausal women and menopausal hormone therapy may be beneficial in reducing this risk.