Long-term success rate of entropion surgery is equivalent in juvenile and adult dogs.

OBJECTIVE: To compare long-term outcomes of lower lid entropion surgery performed in juvenile dogs versus adult dogs and evaluate the success rate of temporary tacking procedures in dogs < 1 year of age. ANIMALS: 116 client-owned dogs. METHODS: A retrospective study was performed evaluating dogs younger than 3 years old diagnosed with primary lower lid entropion between 2010 and 2020. Recurrence of entropion following temporary tacking sutures was evaluated. Surgical outcomes were evaluated of entropion surgery in dogs < and > 1 year of age. RESULTS: 44 dogs with entropion (71 eyes) had a temporary tacking procedure. The entropion resolved in 36.6% of eyes, requiring no further therapy. The median age of dogs successfully treated with a temporary tacking procedure was younger than those that failed. Forty-seven dogs (75 eyes) had entropion surgery at maturity, and 52 dogs (79 eyes) were juvenile. Twenty-seven dogs had temporary tacking procedure prior to surgery, accounting for the difference in number. There was no statistically significant difference in the recurrence rate of entropion between eyes of adult (6/75 [8%]) and juvenile dogs (10/79 [12.7%]) following surgery. CLINICAL RELEVANCE: Entropion surgery in juvenile dogs is not associated with a higher risk of recurrence and need not be delayed until dogs are older than 1 year of age.

Phacoemulsification and intraocular lens implantation in a Canada lynx with phacoclastic uveitis.

A 4-year-old male Canada lynx (Lynx canadensis) was referred to the ophthalmology service at the University of Saskatchewan with a 7-month history of cataract and chronic phacoclastic uveitis secondary to penetrating trauma from a lynx claw. Ophthalmic examination of the right eye revealed a corneal scar, marked aqueous flare, extensive fibrovascular membranes extending from the iris to the lens, anterior and posterior synechiae, immature cataract, and anterior vitritis; the fundus was not visible. Phacoemulsification surgery and intraocular lens implantation using a custom lens of D+46 and 14 mm (An-vision, West Jordan, Utah, USA) was performed. Post-operative medications included sub-conjunctival injections of atropine, cefazolin, and triamcinolone, and oral doxycycline and prednisolone. At the 5-month follow-up, the uveitis was controlled, and a normal fundus was visualized; at 21 mo, the eye remained comfortable and visual. This is the first case report to describe phacoemulsification in a wild felid as a treatment for a traumatic cataract and severe phacoclastic uveitis. Key clinical message: Despite chronic phacoclastic uveitis, phacoemulsification surgery can provide a positive outcome for mature wild felids with traumatic lens rupture, even when topical treatment cannot be administered.

Phacoémulsification et implantation de lentilles intraoculaires chez un lynx du Canada atteint d'uvéite phacoclastique. Un lynx du Canada mâle de 4 ans (Lynx canadensis) a été référé au service d'ophtalmologie de l'University of Saskatchewan avec une histoire de 7 mois de cataracte et d'uvéite phacoclastique chronique secondaire à un traumatisme pénétrant d'une griffe de lynx. L'examen ophtalmique de l'oeil droit a révélé une cicatrice cornéenne, un phénomène de Tyndall marqué, des membranes fibrovasculaires étendues s'étendant de l'iris au cristallin, des synéchies antérieure et postérieure, une cataracte immature et une vitreite antérieure; le fond d'oeil n'était pas visible. Une chirurgie de phacoémulsification et une implantation de lentille intraoculaire à l'aide d'une lentille personnalisée de D+46 et 14 mm (An-vision, West Jordan, Utah, USA) ont été réalisées. Les médicaments postopératoires comprenaient des injections sous-conjonctivales d'atropine, de céfazoline et de triamcinolone, ainsi que de la doxycycline et de la prednisolone par voie orale. Au suivi à 5 mois, l'uvéite était contrôlée et un fond d'oeil normal était visualisé; à 21 mois, l'oeil restait confortable et visuel. Il s'agit du premier rapport de cas décrivant la phacoémulsification chez un félin sauvage comme traitement d'une cataracte traumatique et d'une uvéite phacoclastique sévère.Message clinique clé:Malgré l'uvéite phacoclastique chronique, la chirurgie de phacoémulsification peut donner un résultat positif pour les félins sauvages matures présentant une rupture traumatique du cristallin, même lorsqu'un traitement topique ne peut pas être administré.(Traduit par Dr Serge Messier).

Functional screen of inflammatory bowel disease genes reveals key epithelial functions.

BACKGROUND: Genetic studies have been tremendously successful in identifying genomic regions associated with a wide variety of phenotypes, although the success of these studies in identifying causal genes, their variants, and their functional impacts has been more limited. METHODS: We identified 145 genes from IBD-associated genomic loci having endogenous expression within the intestinal epithelial cell compartment. We evaluated the impact of lentiviral transfer of the open reading frame (ORF) of these IBD genes into the HT-29 intestinal epithelial cell line via transcriptomic analyses. By comparing the genes in which expression was modulated by each ORF, as well as the functions enriched within these gene lists, we identified ORFs with shared impacts and their putative disease-relevant biological functions. RESULTS: Analysis of the transcriptomic data for cell lines expressing the ORFs for known causal genes such as HNF4a, IFIH1, and SMAD3 identified functions consistent with what is already known for these genes. These analyses also identified two major clusters of genes: Cluster 1 contained the known IBD causal genes IFIH1, SBNO2, NFKB1, and NOD2, as well as genes from other IBD loci (ZFP36L1, IRF1, GIGYF1, OTUD3, AIRE and PITX1), whereas Cluster 2 contained the known causal gene KSR1 and implicated DUSP16 from another IBD locus. Our analyses highlight how multiple IBD gene candidates can impact on epithelial structure and function, including the protection of the mucosa from intestinal microbiota, and demonstrate that DUSP16 acts a regulator of MAPK activity and contributes to mucosal defense, in part via its regulation of the polymeric immunoglobulin receptor, involved in the protection of the intestinal mucosa from enteric microbiota. CONCLUSIONS: This functional screen, based on expressing IBD genes within an appropriate cellular context, in this instance intestinal epithelial cells, resulted in changes to the cell's transcriptome that are relevant to their endogenous biological function(s). This not only helped in identifying likely causal genes within genetic loci but also provided insight into their biological functions. Furthermore, this work has highlighted the central role of intestinal epithelial cells in IBD pathophysiology, providing a scientific rationale for a drug development strategy that targets epithelial functions in addition to the current therapies targeting immune functions.

Effect of intracameral epinephrine on heart rate, post-operative ocular hypertension, and long-term outcome following canine phacoemulsification.

PURPOSE: To determine the effect of intrsacameral epinephrine on heart rate, blood pressure, post-operative ocular hypertension, and complications following canine phacoemulsification. PROCEDURES: A prospective, double-blinded, controlled trial was carried out using 30 client-owned dogs undergoing phacoemulsification. Eyes were randomly assigned to a treatment group receiving intracameral (IC) epinephrine (n = 31) or balanced salt solution (n = 25) at the beginning of surgery. Heart rate, post-operative intraocular pressures, and outcomes were compared between treatment groups. RESULTS: No adverse reactions to IC epinephrine or saline were observed. Post-operative ocular hypertension developed at the 2 and/or 4 hours pressure reading in 35% and 46% in the epinephrine and saline groups, respectively (P = .5072). There were 9.7% and 23.1% eyes that developed complications in the IC epinephrine and saline groups, respectively (P = .2373). CONCLUSIONS: Intracameral epinephrine is safe to use, and non-significant decreases in post-operative ocular hypertension and long-term complications were observed.

Chitosan rate of uptake in HEK293 cells is influenced by soluble versus microparticle state and enhanced by serum-induced cell metabolism and lactate-based media acidification.

UNLABELLED: Chitosan is a biocompatible polysaccharide composed of glucosamine and N-acetylglucosamine. The polymer has a unique behavior of fluctuating between soluble chains at pH 6 and insoluble microparticles at pH 7. The purpose of this study was to test the hypothesis that chitosan structure, solubility state, and serum influence the rate of cell uptake. Chitosans with 80% and 95% degree of deacetylation (medium and low viscosity) were tagged with rhodamine and analyzed for particle size, media solubility, and uptake by HEK293 epithelial cells using live confocal microscopy and flow cytometry. In media pH 7.4 with or without 10% serum, chitosans fully precipitated into 0.5 to 1.4 µm diameter microparticles with a slight negative charge. During 24 h of culture in serum-free medium, chitosan particles remained extracellular. In cultures with serum, particles were taken up into intracellular vesicles in a serum dose-dependent manner. Opsonization of chitosan with serum, or replacement of serum by epidermal growth factor (EGF) failed to mediate serum-free chitosan particle uptake. Serum stimulated cells to acidify the media, partly by lactate generation. Media acidified to pH 6.5 by 7 mM lactate maintained 50% of chitosan in the soluble fraction, and led to minor uniform serum-free uptake in small vesicles. CONCLUSION: Media acidification mediates minor in vitro uptake of non-biofouled soluble chitosan chains, while serum-biofouled insoluble chitosan microparticles require sustained serum exposure to generate energy required for macropinocytosis.

The identification of 4,7-disubstituted naphthoic acid derivatives as UDP-competitive antagonists of P2Y14.

A weak, UDP-competitive antagonist of the pyrimidinergic receptor P2RY(14) with a naphthoic acid core was identified through high-throughput screening. Optimization provided compounds with improved potency but poor pharmacokinetics. Acylglucuronidation was determined to be the major route of metabolism. Increasing the electron-withdrawing nature of the substituents markedly reduced glucuronidation and improved the pharmacokinetic profile. Additional optimization led to the identification of compound 38 which is an 8 nM UDP-competitive antagonist of P2Y(14) with a good pharmacokinetic profile.

The C3a anaphylatoxin receptor is a key mediator of insulin resistance and functions by modulating adipose tissue macrophage infiltration and activation.

OBJECTIVE: Significant new data suggest that metabolic disorders such as diabetes, obesity, and atherosclerosis all posses an important inflammatory component. Infiltrating macrophages contribute to both tissue-specific and systemic inflammation, which promotes insulin resistance. The complement cascade is involved in the inflammatory cascade initiated by the innate and adaptive immune response. A mouse genomic F2 cross biology was performed and identified several causal genes linked to type 2 diabetes, including the complement pathway. RESEARCH DESIGN AND METHODS: We therefore sought to investigate the effect of a C3a receptor (C3aR) deletion on insulin resistance, obesity, and macrophage function utilizing both the normal-diet (ND) and a diet-induced obesity mouse model. RESULTS: We demonstrate that high C3aR expression is found in white adipose tissue and increases upon high-fat diet (HFD) feeding. Both adipocytes and macrophages within the white adipose tissue express significant amounts of C3aR. C3aR(-/-) mice on HFD are transiently resistant to diet-induced obesity during an 8-week period. Metabolic profiling suggests that they are also protected from HFD-induced insulin resistance and liver steatosis. C3aR(-/-) mice had improved insulin sensitivity on both ND and HFD as seen by an insulin tolerance test and an oral glucose tolerance test. Adipose tissue analysis revealed a striking decrease in macrophage infiltration with a concomitant reduction in both tissue and plasma proinflammatory cytokine production. Furthermore, C3aR(-/-) macrophages polarized to the M1 phenotype showed a considerable decrease in proinflammatory mediators. CONCLUSIONS: Overall, our results suggest that the C3aR in macrophages, and potentially adipocytes, plays an important role in adipose tissue homeostasis and insulin resistance.

The Kruppel-like transcription factor KLF13 is a novel regulator of heart development.

In humans, congenital heart defects occur in 1-2% of live birth, but the molecular mechanisms and causative genes remain unidentified in the majority of cases. We have uncovered a novel transcription pathway important for heart morphogenesis. We report that KLF13, a member of the Krüppel-like family of zinc-finger proteins, is expressed predominantly in the heart, binds evolutionarily conserved regulatory elements on cardiac promoters and activates cardiac transcription. KLF13 is conserved across species and knockdown of KLF13 in Xenopus embryos leads to atrial septal defects and hypotrabeculation similar to those observed in humans or mice with hypomorphic GATA-4 alleles. Physical and functional interaction with GATA-4, a dosage-sensitive cardiac regulator, provides a mechanistic explanation for KLF13 action in the heart. The data demonstrate that KLF13 is an important component of the transcription network required for heart development and suggest that KLF13 is a GATA-4 modifier; by analogy to other GATA-4 collaborators, mutations in KLF13 may be causative for congenital human heart disease.