Sprouted Innervation into Uterine Transplants Contributes to the Development of Hyperalgesia in a Rat Model of Endometriosis

Endometriosis is an enigmatic painful disorder whose pain symptoms remain difficult to alleviate in large part because the disorder is defined by extrauteral endometrial growths whose contribution to pain is poorly understood. A rat model (ENDO) involves autotransplanting on abdominal arteries uterine segments that grow into vascularized cysts that become innervated with sensory and sympathetic fibers. ENDO rats exhibit vaginal hyperalgesia. We used behavioral, physiological, and immunohistochemical methods to test the hypothesis that cyst innervation contributes to the development of this hyperalgesia after transplant. Rudimentary sensory and sympathetic innervation appeared in the cysts at two weeks, sprouted further and more densely into the cyst wall by four weeks, and matured by six weeks post-transplant. Sensory fibers became abnormally functionally active between two and three weeks post-transplant, remaining active thereafter. Vaginal hyperalgesia became significant between four and five weeks post-transplant, and stabilized after six to eight weeks. Removing cysts before they acquired functional innervation prevented vaginal hyperalgesia from developing, whereas sham cyst removal did not. Thus, abnormally-active innervation of ectopic growths occurs before hyperalgesia develops, supporting the hypothesis. These findings suggest that painful endometriosis can be classified as a mixed inflammatory/neuropathic pain condition, which opens new avenues for pain relief. The findings also have implications beyond endometriosis by suggesting that functionality of any transplanted tissue can be influenced by the innervation it acquires

The Prevalence and Influence of the Combination of Humor and Violence in Super Bowl Commercials

The growing concern over violence in the media has led to vast amounts of research examining the effects of violent media on viewers. An important subset of this research looks at how humor affects this relationship. While research has considered this subset in television programming, almost no research has explored this in the context of advertising. This paper builds on the little research that exists by examining the effects of combining humor and violence, as well as the theoretical approaches that underlie these effects. A content analysis is conducted to identify the prevalence of violence, humor, and the combination of these elements in a longitudinal sample of Super Bowl commercials (2005, 2007, and 2009). Further, we investigate the relationship between the joint occurrence of humor and violence in ads and ad popularity. We conclude that violent acts are rampant in these commercials and that many acts are camouflaged by the simultaneous presence of humor, especially in the most popular ads

A large genome-wide association study of age-related macular degeneration highlights contributions of rare and common variants.

This is the author accepted manuscript. The final version is available from Nature Publishing Group via http://dx.doi.org/10.1038/ng.3448Advanced age-related macular degeneration (AMD) is the leading cause of blindness in the elderly, with limited therapeutic options. Here we report on a study of >12 million variants, including 163,714 directly genotyped, mostly rare, protein-altering variants. Analyzing 16,144 patients and 17,832 controls, we identify 52 independently associated common and rare variants (P < 5 × 10(-8)) distributed across 34 loci. Although wet and dry AMD subtypes exhibit predominantly shared genetics, we identify the first genetic association signal specific to wet AMD, near MMP9 (difference P value = 4.1 × 10(-10)). Very rare coding variants (frequency <0.1%) in CFH, CFI and TIMP3 suggest causal roles for these genes, as does a splice variant in SLC16A8. Our results support the hypothesis that rare coding variants can pinpoint causal genes within known genetic loci and illustrate that applying the approach systematically to detect new loci requires extremely large sample sizes.We thank all participants of all the studies included for enabling this research by their participation in these studies. Computer resources for this project have been provided by the high-performance computing centers of the University of Michigan and the University of Regensburg. Group-specific acknowledgments can be found in the Supplementary Note. The Center for Inherited Diseases Research (CIDR) Program contract number is HHSN268201200008I. This and the main consortium work were predominantly funded by 1X01HG006934-01 to G.R.A. and R01 EY022310 to J.L.H

Peripheral neural sprouting contributes to endo-induced vaginal hyperalgesia in a rat model of endometriosis

Endometriosis, defined by ectopic growths of uterine tissue, is considered an enigma because it is unknown how or even if these abnormal growths contribute to the painful conditions including dysmenorrhea, dyspareunia, and chronic pelvic pain that often accompany the disease. Many clinicians and biomedical scientists assume that the amount of ectopic growth (cysts) predicts the presence or severity of pain symptoms, even though considerable evidence suggests that this assumption is unwarranted. Studies from our laboratory using a rat model of surgically-induced endometriosis (ENDO) demonstrated for the first time that the cysts develop a sensory and sympathetic nerve supply. This discovery gave rise to the hypothesis that this newly-sprouted innervation of the cysts is a significant contributor to the development (i.e., generation) and maintenance of painful symptoms. One of these common symptoms, studied here, is vaginal hyperalgesia (often called dyspareunia in women). The purpose of this dissertation was to use a combination of immunohistochemical, physiological, and behavioral methods to test various aspects of this hypothesis. In the first study, the developmental time course of cyst innervation (sensory and sympathetic) and ENDO-induced vaginal hyperalgesia was examined over a 10 week period post-ENDO. It was found that rudimentary innervation appears within the cysts at 2 weeks post-ENDO, and becomes active at 3 weeks post-ENDO. Between 4 and 5 weeks post-ENDO, vaginal hyperalgesia becomes significant, but is highly variable as the innervation increases and approaches maturity. By 8 to 10 weeks post-ENDO the cyst innervation and hyperalgesia have both matured completely, plateaued and stabilized. Based on these findings, the developmental timeline was divided into three phases: INITIAL (1-2 weeks post-ENDO), TRANSITIONAL (4-6 weeks post-ENDO), and ESTABLISHED (8-10 weeks post-ENDO). In each phase, characteristics of the cyst innervation and vaginal hyperalgesia were found to be as follows: INITIAL, no innervation and no vaginal hyperalgesia; TRANSITIONAL, immature but active innervation and significant but highly variable hyperalgesia; ESTABLISHED, mature innervation and stabilized hyperalgesia both of which varied with the estrous cycle. Then, in each of the three phases, the contribution of the cysts (and their innervation) to ENDO-induced vaginal hyperalgesia was tested, by removing the cysts and assessing the effect on the development and maintenance of the vaginal hyperalgesia. In the TRANSITIONAL phase, the relationship between the severity of ENDO-induced vaginal hyperalgesia and the innervation of the cysts, eutopic uterus, and vaginal canal was also assessed. The effect of cyst removal on ENDO-induced vaginal hyperalgesia in the INITIAL phase prevented the development of vaginal hyperalgesia. In the TRANSITIONAL phase, cyst removal did not significantly alleviate the vaginal hyperalgesia developed prior to cyst-removal, but, prevented its future development. In the ESTABLISHED phase, cyst removal completely alleviated the vaginal hyperalgesia. Further, in the TRANSITIONAL phase, innervation of the cysts (sensory and sympathetic) and innervation of the vaginal canal (sympathetic only) significantly correlated with severity of ENDO-induced vaginal hyperalgesia. Overall, results from these studies strongly support the general hypothesis that the innervation of the cysts contributes to ENDO-induced vaginal hyperalgesia. Specifically, the cyst innervation likely contributes to the development , severity, and maintenance of ENDO-vaginal hyperalgesia. Importantly however, the varying effects of cyst removal suggest that mechanisms by which the innervation operates to contribute to the vaginal hyperalgesia change during its progression through the three phases from peripheral sensitization to peripherally-independent then peripherally-dependent, hormonally-modulated central sensitization. Thus changes, which emerge most clearly in the TRANSITIONAL phase, could help explain the poorly-understood, clinically-challenging issue on how pain transitions from an acute to a chronic problem, not only in endometriosis but also in other chronic pain conditions

A behavioral study of the development of vaginal hyperalgesia and an immunohistochemical study of the development of cyst innervation after transplant surgery.

<p>Note that sensory and sympathetic fibers appear within the cysts two weeks before significant hyperalgesia develops. <i>A</i>. Hyperalgesic severity (post-transplant AUC minus baseline AUC) at different times after transplant surgery in the same rats. Error bars are ± SEM (<i>n</i> = 18). *, differs from two weeks; #, differs from six weeks, <i>p</i><0.05. For the 4 week time point, <i>*t</i> = 0.058. <i>B</i>. Photomicrographs of sensory (CGRP-positive and sympathetic (VMAT2-positive) fibers in the cysts labeled at two, four, six, and ten weeks after transplant surgery (n = 3–5 rats/survival time). Calibration bar is 50 µm for all images.</p

Evans Blue dye extravasation in intact and denervated cysts harvested at different times after transplant surgery.

<p>Note that the cysts' sensory innervation becomes functional (i.e., contributes to EB dye extravasation) three weeks after transplant surgery, which is one week before the hyperalgesia becomes significant (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0031758#pone-0031758-g001" target="_blank">Fig. 1A</a>). *, differs from intact at the same time period, <i>p</i>≤0.05. Within the Intact group: #, differs from the one-week time period, (<i>p</i>≤0.05). Error bars are ± SEM (<i>n</i> = 8–12/group).</p

Effect of cyst removal or sham-cyst removal performed before the cysts acquire mature innervation on the development of vaginal hyperalgesia.

<p><b><i>A</i></b>. This diagram depicts the time-course of experimental testing (baseline, post-transplant) and surgical procedures (1, 2 or 4 weeks after transplant). <b>B</b>–<b>D.</b> The graphs show percent escape response as a function of vaginal distention volume before (baseline, solid line) and 5–10 weeks after transplant surgery (dashed line). Error bars are ± SEM. The inset bar graphs depict the severity of hyperalgesia individually for each rat in that group (as in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0031758#pone-0031758-g001" target="_blank">Fig. 1</a>). Solid bars indicate cyst removal or sham-cyst removal at 1 or 2 weeks post-transplant; the hatched bars indicate cyst removal or sham-cyst removal at 4 weeks post-transplant. <b>B.</b> Pre-innervation cyst removal (1 or 2 weeks post-transplant) prevents the development of hyperalgesia (n = 5). <b>C.</b> When cyst removal is performed just after innervation has become active, but before innervation is mature; i.e., 4 weeks post-transplant, the development of hyperalgesia is prevented in two of three rats. <b>D.</b> In contrast, after sham-cyst removal, regardless of when the surgery is performed relative to the innervation (either 2 or 4 weeks after transplant), hyperalgesia develops in all rats (n = 4).</p

Endocannabinoid involvement in endometriosis

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