Effects of spinal non-viral interleukin-10 gene therapy formulated with d-mannose in neuropathic interleukin-10 deficient mice: Behavioral characterization, mRNA and protein analysis in pain relevant tissues.

Studies show that spinal (intrathecal; i.t.) interleukin-10 (IL-10) gene therapy reverses neuropathic pain in animal models, and co-administration with the mannose receptor (MR; CD206) ligand d-mannose (DM) greatly improves therapeutic efficacy. However, the actions of endogenous IL-10 may be required for enduring pain control observed following i.t. IL-10 gene therapy, potentially narrowing the application of this non-viral transgene delivery approach. Here, we show that i.t. application of naked plasmid DNA expressing the IL-10 transgene co-injected with DM (DM/pDNA-IL-10) for the treatment of peripheral neuropathic pain in IL-10 deficient (IL-10 KO) mice results in a profound and prolonged bilateral pain suppression. Neuropathic pain is induced by unilateral sciatic chronic constriction injury (CCI), and while enduring relief of light touch sensitivity (mechanical allodynia) in both wild type (WT) and IL-10 KO mice was observed following DM/pDNA-IL-10 co-therapy, transient reversal from allodynia was observed following i.t. DM alone. In stably pain-relieved IL-10 KO mice given DM/pDNA-IL-10, mRNA for the IL-10 transgene is detected in the cauda equina and ipsilateral dorsal root ganglia (DRG), but not the lumbar spinal cord. Further, DM/pDNA-IL-10 application increases anti-inflammatory TGF-ß1 and decreases pro-inflammatory TNF mRNA in the ipsilateral DRG compared to allodynic controls. Additionally, DM/pDNA-IL-10 treated mice exhibit decreased spinal pro-inflammatory mRNA expression for TNF, CCL2 (MCP-1), and for the microglial-specific marker TMEM119. Similarly, DM/pDNA-IL-10 treatment decreases immunoreactivity for the astrocyte activation marker GFAP in lumbar spinal cord dorsal horn. Despite transient reversal and early return to allodynia in DM-treated mice, lumbar spinal cord revealed elevated TNF, CCL2 and TMEM119 mRNA levels. Both MR (CD206) and IL-10 receptor mRNAs are increased in the DRG following CCI manipulation independent of injection treatment, suggesting that pathological conditions stimulate upregulation and availability of relevant receptors in critical anatomical regions required for the therapeutic actions of the DM/pDNA-IL-10 co-therapy. Taken together, the current report demonstrates that non-viral DM/pDNA-IL-10 gene therapy does not require endogenous IL-10 for enduring relief of peripheral neuropathic pain and does not require direct contact with the spinal cord dorsal horn for robust and enduring relief of neuropathic pain. Spinal non-viral DM/pDNA-IL-10 co-therapy may offer a framework for the development of non-viral gene therapeutic approaches for other diseases of the central nervous system.

Prenatal alcohol exposure potentiates chronic neuropathic pain, spinal glial and immune cell activation and alters sciatic nerve and DRG cytokine levels.

A growing body of evidence indicates that prenatal alcohol exposure (PAE) may predispose individuals to secondary medical disabilities later in life. Animal models of PAE reveal neuroimmune sequelae such as elevated brain astrocyte and microglial activation with corresponding region-specific changes in immune signaling molecules such as cytokines and chemokines. The aim of this study was to evaluate the effects of moderate PAE on the development and maintenance of allodynia induced by chronic constriction injury (CCI) of the sciatic nerve in adult male rat offspring. Because CCI allodynia requires the actions of glial cytokines, we analyzed lumbar spinal cord glial and immune cell surface markers indicative of their activation levels, as well as sciatic nerve and dorsal root ganglia (DRG) cytokines in PAE offspring in adulthood. While PAE did not alter basal sensory thresholds before or after sham manipulations, PAE significantly potentiated adult onset and maintenance of allodynia. Microscopic analysis revealed exaggerated astrocyte and microglial activation, while flow cytometry data demonstrated increased proportions of immune cells with cell surface major histocompatibility complex II (MHCII) and ß-integrin adhesion molecules, which are indicative of PAE-induced immune cell activation. Sciatic nerves from CCI rats revealed that PAE potentiated the proinflammatory cytokines interleukin (IL)-1ß, IL-6 and tumor necrosis factor-alpha (TNFα) protein levels with a simultaneous robust suppression of the anti-inflammatory cytokine, IL-10. A profound reduction in IL-10 expression in the DRG of PAE neuropathic rats was also observed. Taken together, our results provide novel insights into the vulnerability that PAE produces for adult-onset central nervous system (CNS) pathological conditions from peripheral nerve injury.

Chronic Sciatic Neuropathy in Rat Reduces Voluntary Wheel-Running Activity With Concurrent Chronic Mechanical Allodynia.

BACKGROUND: Animal models of peripheral neuropathy produced by a number of manipulations are assessed for the presence of pathologic pain states such as allodynia. Although stimulus-induced behavioral assays are frequently used and important to examine allodynia (ie, sensitivity to light mechanical touch; von Frey fiber test), other measures of behavior that reflect overall function are not only complementary to stimulus-induced responsive measures, but are also critical to gain a complete understanding of the effects of the pain model on quality of life, a clinically relevant aspect of pain on general function. Voluntary wheel-running activity in rodent models of inflammatory and muscle pain is emerging as a reliable index of general function that extends beyond stimulus-induced behavioral assays. Clinically, reports of increased pain intensity occur at night, a period typically characterized with reduced activity during the diurnal cycle. We therefore examined in rats whether alterations in wheel-running activity were more robust during the inactive phase compared with the active phase of their diurnal cycle in a widely used rodent model of chronic peripheral neuropathic pain, the sciatic nerve chronic constriction injury (CCI) model. METHODS: In adult male Sprague Dawley rats, baseline (BL) hindpaw threshold responses to light mechanical touch were assessed using the von Frey test before measuring BL activity levels using freely accessible running wheels (1 hour/day for 7 sequential days) to quantify the distance traveled. Running wheel activity BL values are expressed as total distance traveled (m). The overall experimental design was after BL measures, rats underwent either sham or CCI surgery followed by repeated behavioral reassessment of hindpaw thresholds and wheel-running activity levels for up to 18 days after surgery. Specifically, separate groups of rats were assessed for wheel-running activity levels (1 hour total/trial) during the onset (within first 2 hours) of either the (1) inactive (n = 8/group) or (2) active (n = 8/group) phase of the diurnal cycle. An additional group of CCI-treated rats (n = 8/group) was exposed to a locked running wheel to control for the potential effects of wheel-running exercise on allodynia. The 1-hour running wheel trial period was further examined at discrete 20-minute intervals to identify possible pattern differences in activity during the first, middle, and last portions of the 1-hour trial. The effect of neuropathy on activity levels was assessed by measuring the change from their respective BLs to distance traveled in the running wheels. RESULTS: Although wheel-running distances between groups were not different at BL from rats examined during either the inactive phase of the diurnal cycle or active phase of the diurnal cycle, sciatic nerve CCI reduced running wheel activity levels compared with sham-operated controls during the inactive phase. In addition, compared with sham controls, bilateral low-threshold mechanical allodynia was observed at all time points after surgical induction of neuropathy in rats with free-wheel and locked-wheel access. Allodynia in CCI compared with shams was replicated in rats whose running wheel activity was examined during the active phase of the diurnal cycle. Conversely, no significant reduction in wheel-running activity was observed in CCI-treated rats compared with sham controls at any time point when activity levels were examined during the active diurnal phase. Finally, running wheel activity patterns within the 1-hour trial period during the inactive phase of the diurnal cycle were relatively consistent throughout each 20-minute phase. CONCLUSIONS: Compared with nonneuropathic sham controls, a profound and stable reduction of running wheel activity was observed in CCI rats during the inactive phase of the diurnal cycle. A concurrent robust allodynia persisted in all rats regardless of when wheel-running activity was examined or whether they ran on wheels, suggesting that acute wheel-running activity does not alter chronic low-intensity mechanical allodynia as measured using the von Frey fiber test. Overall, these data support that acute wheel-running exercise with limited repeated exposures does not itself alter allodynia and offers a behavioral assay complementary to stimulus-induced measures of neuropathic pain.

Hypoxia and Dark Adaptation in Diabetic Retinopathy: Interactions, Consequences, and Therapy.

In diabetes, retinal blood flow is compromised, and retinal hypoxia is likely to be further intensified during periods of darkness. During dark adaptation, rod photoreceptors in the outer retina are maximally depolarized and continuously release large amounts of the neurotransmitter glutamate-an energetically demanding process that requires the highest oxygen consumption per unit volume of any tissue of the body. In complete darkness, even more oxygen is consumed by the outer retina, producing a steep fall in the retinal oxygen tension curve which reaches a nadir at the depth of the mitochondrial-rich rod inner segments. In contrast to the normal retina, the diabetic retina cannot meet the added metabolic load imposed by the dark-adapted rod photoreceptors; this exacerbates retinal hypoxia and stimulates the overproduction of vascular endothelial growth factor (VEGF). The use of nocturnal illumination to prevent dark adaptation, specifically reducing the rod photoreceptor dark current, should ameliorate diabetic retinopathy.

The nitrous oxide "dream" of Cora Gray: a dental anesthesia story of 1884.

In the September 1884 issue of Frank Leslie's Popular Monthly magazine, a fictional dramatic short story was published concerning the dental use of nitrous oxide. Entitled, "Cora Gray," it was written by the well-known American journalist and poet John Whittaker Watson (1815-1848), who authored hundreds of sentimental, tragic and dramatic poems, serials and stories concerning the destitute lives and deaths of downtrodden young women of that time. His greatest poetic effort, "Beautiful Snow," (1869) tells of a young prostitute who freezes to death in a snow bank. Watson, born in New York City, was educated at the University of New York, where he studied medicine. He also developed and used his skills as an engraver, journalist and writer. Watson obviously based his imaginative narrative on his medical knowledge of nitrous oxide and its physical and psychological side effects when inhaled. The story centers around the dreamlike romantic experiences of a 19-year-old female dental patient while she is under the effects of this gas. It explicitly depicts the administration of nitrous oxide and the resulting erotic visions and hallucinations that the young patient experiences. We make reference to other cautionary scientific writings from the late 1800s, in order to point out and clarify the potentially negative repercussions of nitrous oxide when administered to female dental patients without the presence of a third party. The ethics and propriety of anesthesia administration remain as perennial questions in dentistry to this day.

Mutations in the human retinal degeneration slow (RDS) gene can cause either retinitis pigmentosa or macular dystrophy.

Mutations in the RDS gene, which encodes the photoreceptor glycoprotein peripherin, have been sought in families with autosomal dominant retinal dystrophies. A cysteine deletion at codon 118/119 is associated with retinitis pigmentosa in one. Three families with similar macular dystrophy have mutations at codon 172, arginine being substituted by tryptophan in two and by glutamine in one. A stop sequence at codon 258 exists in a family with adult vitelliform macular dystrophy. These findings demonstrate that both retinitis pigmentosa and macular dystrophies are caused by mutations in RDS and that the functional significance of certain amino-acids in peripherin-RDS may be different in cones and rods.

Short-term effects of a high-severity summer wildfire on conifer forest moth (Lepidoptera) communities in New Mexico, USA.

Forest fires in North America are becoming larger in area and burning with higher severity as a result of climate change and land management practices. High-severity, stand-replacement fires can inflict major changes to forest insect communities, potentially extirpating many species through altered post-fire habitat resources. We assessed forest-dwelling macrolepidopteran moth communities in mixed conifer and ponderosa pine forests during the first year after the 2011 Las Conchas fire in New Mexico, USA. We deployed blacklight traps in replicated burned and unburned stands during June, July, and August in 2012. We collected 9,478 individuals, representing 211 species and 8 families. Noctuidae (124 species) and Geometridae (53) comprised the majority of the taxa, followed by Erebidae (21), Sphingidae (5), Notodontidae (3), Lasiocampidae (2), Saturniidae (2), and Drepanidae (1). Moth communities (species composition and abundances) in each forest type (mixed conifer vs. ponderosa pine) were statistically distinguishable, but shared 56.4% (119) of observed species. Overall, compared to unburned forests, post-fire moth communities in both forest types had significantly lower numbers of individuals, species richness and diversity, and lower evenness in ponderosa pine forests. As expected, categorizing moth taxa by larval host plant taxa revealed that reductions of moth populations following fire were associated with the elimination or reduction of available larval host plants (particularly conifers, oaks, and junipers). We predict that future moth community succession will likely parallel the overall transformation from a forested landscape to a montane meadow/grassland ecosystem, with continued reduction in tree-feeding species and increasing dominance by forb/grass-feeding species.

The pathogenesis of early retinal changes of diabetic retinopathy.

Recent successful trials of antibodies to vascular endothelial growth factor (VEGF) in diabetic retinopathy implicate this cytokine as a major cause of diabetic retinopathy (DR) and diabetic macular oedema (DME). The mechanisms which cause VEGF to be over-expressed to cause the vasculopathy are not entirely clear. This review explores the earliest changes to the retina in DR and the factors that predispose or prevent DR, including sleep apnoea, receptor degenerations laser treatment and VEGF polymorphism. The review also presents the evidence that retinal hypoxia, existing in the earliest stages, causes DR. This hypoxia is much increased by dark adaptation, indicating a new and possibly superior therapy.

The 1790 desecration of John Milton's bones, hair and teeth in a London church.

John Milton (1608-1674), England's epic poet and a champion of civil and religious liberties, has gone down in history as one of the world's greatest philosophers, scholars and authors. Although by age 44 he had become totally blind, he steadfastly continued to write and eventually composed his poetic masterpieces, "Paradise Lost" and "Paradise Regained." In 1674, John Milton died of kidney failure at age 66. He was buried in the "chancel under the clerk's desk" in the parish church of St. Giles-without-Cripplegate (London). Exhumed 116 years after his burial, Milton's corpse received "obscene and scandalous treatment", as portions of his bones, hair and teeth were removed, examined and indiscriminately dispersed to others.

Retail promotions and perceptions of R.J. Reynolds' novel dissolvable tobacco in a US test market.

BACKGROUND: With declining cigarette sales, tobacco manufacturers have been developing and marketing new smokeless products, such as R. J. Reynolds' dissolvable tobacco, Camel Sticks, Strips and Orbs. This study assessed the availability, price and point-of-purchase promotional strategies for Camel Dissolvables, and investigated consumer awareness, interest and perception of these products in the Indiana test market. METHODS: An exploratory retail audit of point-of-purchase promotions was conducted in a random sample of retailers from 6 store categories (n = 81) in the test market area. Data included: store type, location, product placement, forms/flavors carried, price, types and locations of advertisements and promotions, and ad messages. An Awareness-Attitude-Usage (AAU) survey was used to gauge consumer awareness and knowledge of tobacco products including Camel Dissolvables. Respondents were shown promotional materials from a package onsert and perceptions and interest in the Camel Dissolvables were assessed. An Intended Target Survey (ITS) compared subjects' perceptions of ad targets for several non-tobacco products, as well as Camel Snus, Camel No. 9 and Camel Dissolvables. Respondents were asked to identify each ad's intended target category, perceived targetedness, and purchase intent. RESULTS: The products were carried by 46% of stores, most frequently gas stations (100%) and convenience stores (75%). They were shelved near smokeless tobacco (70%), cigarettes (25%) or candy (5%). Prices ranged from $3.59 -$4.19 per package; most stores carried at least 1 promotional item. Ad messages included: "Dissolvable Tobacco" (60%). "Free Trial" (24%), "Special Price" (24%), "What's Your Style?" (22%). At 14% of stores, free trial packs of Camel Dissolvables were offered with another Camel purchase. Awareness was reported by 42% of respondents (n = 243), and trial by 3%. Consumer interest was very low, but younger respondents (< 40 years) were more familiar with Camel Dissolvables (60% vs. 45% for those > 40 years, p < .01). Males, as well as current and former smokers had higher rates of interest and trial; only 1% of never smokers reported trial. In the ITS, only for the 3 tobacco product ads, was perceived targetedness for smokers significantly higher than for non-smokers. Smokers and nonsmokers perceived that the ads targeted smokers. CONCLUSIONS: Current retail promotional strategies for Camel Sticks, Strips & Orbs appear to be targeting a select audience, primarily current smokers. Overall, consumer awareness, interest and trial were low.

The unicorn and the narwhal: a tale of the tooth.

From as far back as the early Roman era, detailed descriptions and artistic images of the unicorn have been imagined. In Europe, the Mediterranean and the Far East, this creature became accepted as real. Although such an animal had never actually been seen, it was said to have the appearance of a horse, the beard and cloven hooves of a goat, the tail of a lion, and a single, spear-like horn protruding from the center of its forehead. The unicorn myth originated shortly after unidentified horn-like objects, from six to ten feet long, began to appear in ancient European marketplaces. Physicians quickly ascribed outlandish healing properties to these horns, pulverizing them for the treatment of various diseases. In 1638, a Danish zoologist challenged these claims and provided evidence that the so-called "unicorn horn" was actually the tusk (tooth) of the narwhal from the Arctic seas. However, his findings remained unheeded for the next 100 years. This paper explains how the myth of the unicorn and the purported powers contained within the substance of its magical horn evolved from the ages-old spoils of the Arctic fishing trade, where the narwhal was primarily speared for food and secondarily, for the export value of its ivory tusk. This unusually long, pointed and protruding single tooth, which breaks through the upper lip and the left jaw of each male narwhal, became a highly valuable commodity over the following centuries, due to its believed supernatural origins and curative qualities.

"The old family toothbrush": Stotesbury's whimsy.

Edward Townsend Stotesbury (1849-1938), a prominent Philadelphia tycoon, made his millions as an investment banker and financier. In 1927, his fortune was estimated to be $100 million. In 1912, at age 63, Stotesbury, then a widower for over thirty years, married a captivating Chicago widow, Eva Roberts Cromwell. As a "glittering socialite," Cromwell had an extraordinary sense of personal theatre, a keen eye for beauty and a calculating mind. Throughout the 1920s and 30s, the couple built three lavish palatial estates. At these mansions, they entertained on an imposing scale. Stotesbury especially enjoyed entertaining his guests with recollections of the events he experienced as a Civil War drummer boy. To conclude his program, he shared this lively tune: "The Old Family Toothbrush That Hung by the Sink."

John Peter Zenger's fight for freedom of the press: halitosis exposed.

In the fall of 1732, William Cosby, a member of the British aristocracy, became the colonial governor of New York. As an incompetent, tyrannical and self-serving ruler, he quickly alienated the citizens of New York and, as a result, a core group instigated a broad-based popular uprising against him. In November 1733, John Peter Zenger (1697-1746) published his first issue of the New York Weekly Journal, which would soon become a political organ of those who attempted to bring Cosby down. Zenger's Journal contained particularly offensive attacks on Governor Cosby. They included biting personal jabs, which were judged as being quite inappropriate. The governor was described as having "loathsome false teeth and an unclean mouth." Circulating first-hand reports stated that he possessed a chronic case of the most offensive halitosis. In November 1734, Zenger was arrested and jailed for seditious libel. Fortunately for Zenger, he was defended by the colony's most prominent attorney, Andrew Hamilton. One day later, after only ten minutes of jury deliberation, Zenger was found not guilty. This famous first victory, later called the "germ of American freedom", paved the way to First Amendment Rights in America.

Your tooth it ain't so purty but it's gen-u-ine: Cole Porter's comic song "Snagtooth Gertie".

Cole Porter (1891-1964), an American musical icon, has been internationally acknowledged as one of the most cosmopolitan, sophisticated, elegant and influential composer-lyricists of the 20th century. Born in Indiana and raised in wealth and privilege, he attended both Yale and Harvard universities where he honed his song-writing skills. In his 800 tunes, many of which became smash hits, he blended fresh, witty, urbane and colloquial lyrics with catchy, singable melodies. He composed musical scores for both Broadway stage and Hollywood movies, but focused on the former. While Porter enjoyed penning comic songs, he rarely had them published. In 1946, Cole wrote the music and lyrics for a Broadway show entitled Around the World in Eighty Days. Deemed a failure, it closed after only 75 performances. One light-hearted ditty, "Snagtooth Gertie", which he composed for the show, was never used. The lyrics are presented for the first time in this paper.

Mosquitoes, flies and dental cavities: Dr. Howard Riley Raper's public campaign to prevent toothache.

Dr. Howard Riley Raper (1886-1978) was an early oral health pioneer and dental roentgenology faculty member of the Indiana Dental College (IDC) who single-handedly introduced key concepts in radiology to dentistry. Due to his efforts, IDC became in 1910-11 the first dental school to have a regular course in dental radiology. Virtually all American dental schools soon added this subject to their regular curriculum. Raper's text, Elementary and Dental Radiography (1913) became the first comprehensive student textbook of dental X-ray diagnosis. In his 1933 Blue Book entitled, The New Aim in the Care of the Teeth, Raper elaborated upon his mission to prevent caries, by comparing the insidious damages of tooth decay with the threat of insect-borne disease.

Dental engravings created after works by Adriaen Brouwer: 17th century Flemish genre artist.

Adriene Brouwer (1605/6-1638), a Flemish genre and landscape painter, was prototypical Bohemian. An artist who lived and acted with disregard for conventional rules of behavior, he produced works which often shocked the staid citizenry of his times. This article examines two dentally-oriented etchings crafted by Brouwer. "The Operation" portrays separate scenes in which two lowly tradesmen--a chiropodist and a peasant toothdrawer--are simultaneously at work in the opposite ends of the same room. A second etching, "Peasant Doctoring the Foot of Another Peasant", accentuates the edentulous facial features, both of the suffering patient and of the operator.