Extensive intraocular melanoma with secondary glaucoma in a 15-month-old Thoroughbred filly.

A 15-month-old, grey, Thoroughbred filly presented for investigation of a 6-week history of corneal oedema and blepharospasm on the right eye (OD). The filly was otherwise healthy. Following ophthalmic examination, glaucoma on the OD was diagnosed. A space occupying mass within the anterior chamber was documented on transpalpebral ultrasonographic examination. This mass obliterated most of the anterior intraocular structures on the peripheral nasal side (corneal endothelium and drainage angle), leading to secondary glaucoma. After systemic and topical treatment addressing secondary glaucoma, the corneal oedema reduced. The mass was visualised as an irregularly rounded brown structure associated with the iris on the peripheral nasal side of the anterior chamber. Given the filly's signalment, location and appearance of the mass, a tentative diagnosis of intraocular melanoma was made and enucleation was performed. Histopathological evaluation of the globe revealed solid sheets of heavily pigmented melanocytic cells, disrupting the normal ciliary body architecture and extending into the iris and subretinal. The cells were pleomorphic, polyhedral to round with occasional spindle-shaped cells, and contained moderate to large amounts of granular black-brown pigment (melanin). The iridal component expanded into the anterior chamber, with cells directly opposed to Descemet's membrane, with loss of the endothelium and expanding and occluding the filtration angle in this area. The lesion infiltrated locally into the edge of the sclera, but did not extend through the sclera, though occasional perivascular clusters of melanophages were observed within the scleral stroma adjacent to the optic nerve. Diagnosis of a uveal melanocytic neoplasm was confirmed, with characteristics similar to only one reported case . This is a unique case of a rapidly growing, invasive, uveal melanoma in a young horse. Intraocular melanoma should be considered as a differential diagnoses for glaucoma in grey horses, regardless of the age and absence of melanocytic skin lesions.

Breastfeeding Following Spinal Cord Injury: Consumer Guide for Mothers.

The World Health Organization (WHO) recommends that infants be breastfed exclusively for the first 6 months of age. However, there are few resources available on the effects a spinal cord injury (SCI) can have for breastfeeding mothers. It is difficult to find information to address the unique challenges women with SCI experience when planning or trying to breastfeed. Our international team, including women with SCI, health care providers, and SCI researchers, aims to address the information gap through the creation of this consumer guide. The purpose of this consumer guide is to share the most common issues women with SCI experience during breastfeeding and provide information, practical suggestions, recommendations, and key resources in lay language. General information about breastfeeding is available on the internet, in books, or from friends and health care providers. We do not intend to repeat nor replace general breastfeeding information or medical advice. Breastfeeding for mothers with SCI is complex and requires a team of health care providers with complementary expertise. Such a team may include family physician, obstetrician, physiatrist, neurologist, occupational and physical therapist, lactation consultant, midwife, and psychologist. We hope this consumer guide can serve as a quick reference guide for mothers with SCI planning of trying to breastfeed. This guide will also be helpful to health care providers as an educational tool.

Motherhood after Spinal Cord Injury: Breastfeeding, Autonomic Dysreflexia, and Psychosocial Health: Clinical Practice Guidelines.

The World Health Organization (WHO) recommends that children be breastfed exclusively for the first 6 months of age. This recommendation may prove challenging for women with spinal cord injury (SCI) who face unique challenges and barriers to breastfeeding due to the impact of SCI on mobility and physiology. Tailored provision of care from health care professionals (HCPs) is important in helping women navigate these potential barriers. Yet, HCPs often lack the confidence and SCI-specific knowledge to meet the needs of mothers with SCI. An international panel of clinicians, researchers, consultants, and women with lived experience was formed to create an accessible resource that can address this gap. A comprehensive survey on breastfeeding complications, challenges, resources, and quality of life of mothers with SCI was conducted, along with an environmental scan to evaluate existing postpartum guidelines and assess their relevance and usability as recommendations for breastfeeding after SCI. Building on this work, this article provides evidence-based recommendations for HCPs, including but not limited to general practitioners, obstetricians, pediatricians, physiatrists, lactation consultants, nurses, midwives, occupational therapists, and physiotherapists who work with prospective and current mothers with SCI.

Ex Vivo Corneal Organ Culture Model for Wound Healing Studies.

The cornea has been used extensively as a model system to study wound healing. The ability to generate and utilize primary mammalian cells in two dimensional (2D) and three dimensional (3D) culture has generated a wealth of information not only about corneal biology but also about wound healing, myofibroblast biology, and scarring in general. The goal of the protocol is an assay system for quantifying myofibroblast development, which characterizes scarring. We demonstrate a corneal organ culture ex vivo model using pig eyes. In this anterior keratectomy wound, corneas still in the globe are wounded with a circular blade called a trephine. A plug of approximately 1/3 of the anterior cornea is removed including the epithelium, the basement membrane, and the anterior part of the stroma. After wounding, corneas are cut from the globe, mounted on a collagen/agar base, and cultured for two weeks in supplemented-serum free medium with stabilized vitamin C to augment cell proliferation and extracellular matrix secretion by resident fibroblasts. Activation of myofibroblasts in the anterior stroma is evident in the healed cornea. This model can be used to assay wound closure, the development of myofibroblasts and fibrotic markers, and for toxicology studies. In addition, the effects of small molecule inhibitors as well as lipid-mediated siRNA transfection for gene knockdown can be tested in this system.

Isolation and Characterization of Cutaneous Epithelial Stem Cells.

The outer layer of mammalian skin is a multilayered epithelium that perpetually renews multiple differentiated lineages. During homeostasis, the maintenance of skin epithelial turnover is ensured by regionalized populations of stem cells that largely remain dedicated to distinct epithelial lineages including squamous, follicular, sebaceous, Merkel, and sweat glands. Cutting edge developments in this field have focused on: (1) stem cell activation cues derived from a number of extrinsic sources including neurons, dermal fibroblasts and adipocyte, and immune cells; and (2) characterization of epithelial stem cell homeostasis via hierarchical versus stochastic paradigms. The techniques outlined in this chapter are designed to facilitate such studies and describe basic procedures for cutaneous stem cell isolation and purification, which are based on leveraging their unique expression of surface proteins for simultaneous targeting and purifying of multiple subpopulations in adult skin. In addition, protocols for assessment of in vitro and ex vivo progenitor capacity as well as techniques to visualize progenitor populations in whole skin are discussed.

Identification of Affective State Change in Adults With Aphasia Using Speech Acoustics.

Purpose: The current study aimed to identify objective acoustic measures related to affective state change in the speech of adults with post-stroke aphasia. Method: The speech of 20 post-stroke adults with aphasia was recorded during picture description and administration of the Western Aphasia Battery-Revised (Kertesz, 2006). In addition, participants completed the Self-Assessment Manikin (Bradley & Lang, 1994) and the Stress Scale (Tobii Dynavox, 1981-2016) before and after the language tasks. Speech from each participant was used to detect a change in affective state test scores between the beginning and ending speech. Results: Machine learning revealed moderate success in classifying depression, minimal success in predicting depression and stress numeric scores, and minimal success in classifying changes in affective state class between the beginning and ending speech. Conclusions: The results suggest the existence of objectively measurable aspects of speech that may be used to identify changes in acute affect from adults with aphasia. This work is exploratory and hypothesis-generating; more work will be needed to make conclusive claims. Further work in this area could lead to automated tools to assist clinicians with their diagnoses of stress, depression, and other forms of affect in adults with aphasia.

The deubiquitylase USP10 regulates integrin ß1 and ß5 and fibrotic wound healing.

Scarring and fibrotic disease result from the persistence of myofibroblasts characterized by high surface expression of αv integrins and subsequent activation of the transforming growth factor ß (TGFß) proteins; however, the mechanism controlling their surface abundance is unknown. Genetic screening revealed that human primary stromal corneal myofibroblasts overexpress a subset of deubiquitylating enzymes (DUBs), which remove ubiquitin from proteins, preventing degradation. Silencing of the DUB USP10 induces a buildup of ubiquitin on integrins ß1 and ß5 in cell lysates, whereas recombinant USP10 removes ubiquitin from these integrin subunits. Correspondingly, the loss and gain of USP10 decreases and increases, respectively, αv/ß1/ß5 protein levels, without altering gene expression. Consequently, endogenous TGFß is activated and the fibrotic markers alpha-smooth muscle actin (α-SMA) and cellular fibronectin (FN-EDA) are induced. Blocking either TGFß signaling or cell-surface αv integrins after USP10 overexpression prevents or reduces fibrotic marker expression. Finally, silencing of USP10 in an ex vivo cornea organ culture model prevents the induction of fibrotic markers and promotes regenerative healing. This novel mechanism puts DUB expression at the head of a cascade regulating integrin abundance and suggests USP10 as a novel antifibrotic target.

Autophagy and Mitochondrial Dysfunction in Tenon Fibroblasts from Exfoliation Glaucoma Patients.

PURPOSE: To test the hypothesis that autophagy dysfunction is involved in exfoliation syndrome (XFS), a systemic disorder of extracellular elastic matrices that causes a distinct form of human glaucoma. METHODS: Fibroblasts derived from tenon tissue discards (TFs) from filtration surgery to relieve intraocular pressure in XFS patients were compared against age-matched TFs derived from surgery in primary open-angle glaucoma (POAG) patients or from strabismus surgery. Differential interference contrast light, and electron microscopy were used to examine structural cell features. Immunocytochemistry was used to visualize LOXL1 and Fibulin-5, lysosomes, endosomes, Golgi, and microtubules. Light scatter, Cyto-IDTM and JC1 flow cytometry were used to measure relative cell size, autophagic flux rate and mitochondrial membrane potential (MMPT), respectively. Enhanced autophagy was induced by serum withdrawal. RESULTS: In culture, XFS-TFs were 1.38-fold larger (by light scatter ratio, p = 0.05), proliferated 42% slower (p = 0.026), and were morphologically distinct in 2D and 3D culture compared to their POAG counterparts. In extended 3D cultures, XFS-TFs accumulated 8-10 times more Fibulin-5 than the POAG-TFs, and upon serum withdrawal, there were marked deficiencies in relocation of endosomes and lysosomes to the perinuclear area. Correspondingly, the XFS-TFs displayed significant accumulation of the autophagasome marker LC3 II (3.9 fold increase compared to POAG levels, p = 0.0001) and autophagic flux rate as measured by Cyto-ID dye was 53% lower in XFS-TFs than in POAG-TFs (p = 0.01), indicating reduced clearance of autophagasomes. Finally the percent of cells with diminished MMPT was 3-8 times larger in the XFS-TFs than in POAG-TFs (p = 0.02). CONCLUSIONS: Our results provide for the first time a link between XFS pathology to autophagy dysfunction, a major contributor to multiple age related diseases systemically throughout the body, in the brain and in the retina. A diminished capacity for degradation of denatured protein and aging cellular organelles may underpin the development of extracellular protein aggregates in XFS.

Expression of insulin-like growth factor 2 receptor in corneal keratocytes during differentiation and in response to wound healing.

PURPOSE: Insulin-like growth factor 2 receptor (IGF2R) associates with ligands that influence wound healing outcomes. However, the expression pattern of IGF2R and its role in the cornea is unknown. METHODS: Human keratocytes were isolated from donor corneas. Fibroblasts (fibroblast growth factor 2 [FGF2]-treated) or myofibroblasts (TGF-ß1-treated) were analyzed for IGF2R and α-smooth muscle actin (α-SMA) expression by Western blotting and immunolocalization. Mouse corneas were wounded in vivo and porcine corneas ex vivo. The IGF2R and α-SMA protein expression were visualized and quantified by immunohistochemistry. The IGF2R gene expression in human corneal fibroblasts was knocked-down with targeted lentiviral shRNA. RESULTS: The IGF2R is expressed in epithelial and stromal cells of normal human, mouse, and porcine corneas. The IGF2R increases (11.2 ± 0.4-fold) in the epithelial and (11.7 ± 0.9-fold) stromal layers of in vivo wounded mouse corneas. Double-staining with α-SMA- and IGF2R-specific antibodies reveals that IGF2R protein expression is increased in stromal myofibroblasts in the wounded cornea relative to keratocytes in the normal cornea (11.2 ± 0.8-fold). Human primary stromal keratocytes incubated with FGF2 or TGF-ß1 in vitro demonstrate increased expression (2.0 ± 0.4-fold) of IGF2R in myofibroblasts relative to fibroblasts. Conversion of IGF2R shRNA-lentiviral particle transduced corneal fibroblasts to myofibroblasts reveals a dependence on IGF2R expression, as only 40% ± 10% of cells transduced converted to myofibroblasts compared to 86% ± 3% in control cells. CONCLUSIONS: The IGF2R protein expression is increased during corneal wound healing and IGF2R regulates human corneal fibroblast to myofibroblast differentiation.

Sexual selection is influenced by both developmental and adult environments.

Sexual selection is often assumed to be strong and consistent, yet increasing research shows it can fluctuate over space and time. Few experimental studies have examined changes in sexual selection in response to natural environmental variation. Here, we use a difference in resource quality to test for the influence of past environmental conditions and current environmental conditions on male and female mate choice and resulting selection gradients for leaf-footed cactus bugs, Narnia femorata. We raised juveniles on natural high- and low-quality diets, cactus pads with and without ripe cactus fruits. New adults were again assigned a cactus pad with or without fruit, paired with a potential mate, and observed for mating behaviors. We found developmental and adult encounter environments affected mating decisions and the resulting patterns of sexual selection for both males and females. Males were not choosy in the low-quality encounter environment, cactus without fruit, but they avoided mating with small females in the high-quality encounter environment. Females were choosy in both encounter environments, avoiding mating with small males. However, they were the choosiest when they were in the low-quality encounter environment. Female mate choice was also context dependent by male developmental environment. Females were more likely to mate with males that had developed on cactus with fruit when they were currently in the cactus with fruit environment. This pattern disappeared when females were in the cactus without fruit environment. Altogether, these results experimentally demonstrate context-dependent mate choice by both males and females. Furthermore, we demonstrate that simple, seasonal changes in resources can lead to fluctuations in sexual selection.

Assessing the accuracy of pregnant women in recalling pre-pregnancy weight and gestational weight gain.

OBJECTIVE: Maternity care providers can use pre-pregnancy weight (PPW) and gestational weight gain (GWG) as markers for difficult delivery, and frequently obtain this information directly from the patient. The goal of this study was to determine whether women report their PPW and GWG correctly at the end of pregnancy. METHODS: We performed a prospective cohort study of 189 women delivering between June 1, 2011, and July 31, 2011, at the Saint John Regional Hospital or the Moncton Hospital in New Brunswick. Self- reported PPW and GWG were compared with measured weights obtained from the antenatal chart and upon presentation for delivery. Patient characteristics, BMI classification, and accuracy and degree of error in recall were assessed. RESULTS: The majority of respondents were under 30 years of age (63.4%) and were delivering at term (96.3%). Ninety women (47.6%) were having their first baby. A record of weight measured in the first trimester was available for 98 respondents (51.9%); using this information, 44 women (44.9%) were determined to be overweight or obese at delivery. Approximately one third of women with a normal BMI were not able to recall their PPW or GWG accurately (± 1 kg). Among all BMI classes, there was a consistent pattern of under-reporting of PPW (by a mean of 1.52 kg) and over-reporting of GWG (by a mean of 1.61 kg), but several extreme outliers were identified. CONCLUSION: At the time of delivery, under-reporting of PPW and over-reporting of GWG are common and difficult to predict. Maternity care providers should be aware of this discrepant reporting of PPW and GWG and recognize the implications for intrapartum management and postpartum weight loss.

Objectif : Les fournisseurs de soins de maternité peuvent utiliser le poids prégrossesse (PPG) et le gain pondéral gestationnel (GPG) à titre de marqueurs pour ce qui est de l'accouchement difficile; de plus, ils peuvent fréquemment obtenir cette information directement de la patiente. Cette étude avait pour objectif de déterminer si les femmes signalent leur PPG et leur GPG correctement à la fin de la grossesse. Méthodes : Nous avons mené une étude de cohorte prospective auprès de 189 femmes ayant accouché entre le 1er juin 2011 et le 31 juillet 2011, au Saint John Regional Hospital ou au Moncton Hospital au Nouveau-Brunswick. Le PPG et le GPG autosignalés ont été comparés aux mesures du poids issues du dossier prénatal et effectuées à l'arrivée de la patiente à l'hôpital pour l'accouchement. Les caractéristiques de la patiente, la classification IMC et la précision et le degré d'erreur en ce qui concerne le signalement des poids ont été évalués. Résultats : La majorité des répondantes avaient moins de 30 ans (63,4 %) et accouchaient à terme (96,3 %). Quatre-vingt-dix femmes (47,6 %) en étaient à leur premier accouchement. Le poids mesuré au cours du premier trimestre avait été consigné en ce qui concerne 98 répondantes (51,9 %); en fonction de ces données, nous avons déterminé que 44 femmes (44,9 %) présentaient une surcharge pondérale ou étaient obèses au moment de l'accouchement. Près du tiers des femmes présentant un IMC normal n'étaient pas en mesure de se rappeler de leur PPG ou de leur GPG avec précision (± 1 kg). Dans toutes les classes d'IMC, nous avons constaté une tendance stable quant au sous-signalement du PPG (de 1,52 kg, en moyenne) et au sursignalement du GPG (de 1,61 kg, en moyenne); cependant, plusieurs valeurs extrêmement aberrantes ont été identifiées. Conclusion : Au moment de l'accouchement, le sous-signalement du PPG et le sursignalement du GPG sont courants et difficiles à prédire. Les fournisseurs de soins de maternité devraient prendre conscience de ces divergences quant signalement du PPG et du GPG, et en reconnaître les conséquences en matière de prise en charge intrapartum et de perte de poids postpartum.

TRPV1 potentiates TGFß-induction of corneal myofibroblast development through an oxidative stress-mediated p38-SMAD2 signaling loop.

Injuring mouse corneas with alkali causes myofibroblast expression leading to tissue opacification. However, in transient receptor potential vanilloid 1 channel (TRPV1-/-) knockout mice healing results in transparency restoration. Since TGFß is the primary inducer of the myofibroblast phenotype, we examined the mechanism by which TRPV1 affects TGFß-induced myofibroblast development. Experiments were performed in pig corneas and human corneal fibroblasts (HCFs). Immunohistochemical staining of α-smooth muscle actin (α-SMA) stress fibers was used to visualize myofibroblasts. Protein and phosphoprotein were determined by Western blotting. siRNA transfection silenced TRPV1 gene expression. Flow cytometry with a reactive oxygen species (ROS) reporting dye analyzed intracellular ROS. [Ca2+]I was measured by loading HCF with fura2. In organ cultured corneas, the TRPV1 antagonist capsazepine drastically reduced by 75% wound-induced myofibroblast development. In HCF cell culture, TGF-ß1 elicited rapid increases in Ca2+ influx, phosphorylation of SMAD2 and MAPKs (ERK1/2, JNK1/2 and p38), ROS generation and, after 72 hrs myofibroblast development. SMAD2 and p38 activation continued for more than 16 h, whereas p-ERK1/2 and p-JNK1/2 waned within 90 min. The long-lived SMAD2 activation was dependent on activated p38 and vice versa, and it was essential to generate a > 13-fold increase in α-SMA protein and a fully developed myofibroblast phenotype. These later changes were markedly reduced by inhibition of TRPV1 or reduction of the ROS generation rate. Taken together our results indicate that in corneal derived fibroblasts, TGFß- induced myofibroblast development is highly dependent on a positive feedback loop where p-SMAD2-induced ROS activates TRPV1, TRPV1 causes activation of p38, the latter in turn further enhances the activation of SMAD2 to establish a recurrent loop that greatly extends the residency of the activated state of SMAD2 that drives myofibroblast development.

Conspecific and heterospecific cues override resource quality to influence offspring production.

Animals live in an uncertain world. To reduce uncertainty, animals use cues that can encode diverse information regarding habitat quality, including both non-social and social cues. While it is increasingly appreciated that the sources of potential information are vast, our understanding of how individuals integrate different types of cues to guide decision-making remains limited. We experimentally manipulated both resource quality (presence/absence of cactus fruit) and social cues (conspecific juveniles, heterospecific juveniles, no juveniles) for a cactus-feeding insect, Narniafemorata (Hemiptera: Coreidae), to ask how individuals responded to resource quality in the presence or absence of social cues. Cactus with fruit is a high-quality environment for juvenile development, and indeed we found that females laid 56% more eggs when cactus fruit was present versus when it was absent. However, when conspecific or heterospecific juveniles were present, the effects of resource quality on egg numbers vanished. Overall, N. femorata laid approximately twice as many eggs in the presence of heterospecifics than alone or in the presence of conspecifics. Our results suggest that the presence of both conspecific and heterospecific social cues can disrupt responses of individuals to environmental gradients in resource quality.

The development and impact of insomnia on cancer survivors: a qualitative analysis.

OBJECTIVES: To conduct the first qualitative analysis of the development and impact of insomnia on a cohort of cancer survivors. METHODS: Twenty-one cancer survivors with a history of chronic insomnia contributed to four focus groups held at the University of Glasgow Sleep Research Centre. Participants' perceptions of the onset, evolution and effects of insomnia were elicited and qualitatively explored using content analysis. RESULTS: Most participants reported insomnia onset following cancer diagnosis. Participants who had a pre-existing insomnia reported that cancer diagnosis significantly aggravated their sleep complaint. Active cancer treatment was a major contributor to poor sleep quality due to the disruption of normal daily routines. This poor sleep pattern became persistent once active treatment had ceased and participants reported becoming particularly concerned about their sleep when they were discharged into follow-up cancer care. The impact of insomnia was significant for all participants in the study and six major areas emerged as being particularly affected; mood, physical health, relationships, sleep quality, sleep-related behaviour and cognition. CONCLUSIONS: The majority of cancer survivors in this study developed disturbed sleep as a result of cancer diagnosis and their sleep disruption was exacerbated by active cancer treatment. Insomnia also had a significant impact upon quality of life and these effects persisted long beyond the cessation of active anti-cancer therapy. Early identification of insomnia symptoms in cancer care settings must be a priority to ensure that sleep disturbance is not overlooked or poorly managed.