Topical applications of allogeneic adipose-derived mesenchymal stem cells ameliorate the canine keratoconjunctivitis sicca.

BACKGROUND: Canine keratoconjunctivitis sicca (KCS) is predominantly an immune-mediated disease. Current therapy of canine KCS is mainly by immunosuppressant, but the effectiveness was limited in some patients. In the past few years, some studies showed the results of the use of mesenchymal stem cells in treating canine KCS via periocular injections. However, the periocular injection procedure requires sedation or general anesthesia, and may lead to iatrogenic or incidental injury during the injection process. The aim of this study was to investigate the efficacy of topical allogenic canine adipose-derived mesenchymal stem cells (cAD-MSCs) in clinical patients of canine KCS. RESULTS: The cAD-MSCs used in this study were characterized for their capability of tri-lineage differentiation and immunomodulatory properties. In addition, preparation methods for eye drops of cAD-MSCs was developed and its optimal preservation was tested. The canine KCS patients were recruited for clinical trial and divided into two groups based on their history of previous treatment. All patients received topical cAD-MSCs treatment once per week for 6 consecutive weeks and complete ophthalmic examinations were performed 1 week before treatment (week 0) and at 3rd, 6th, 9th weeks, respectively. The results showed that the quantity and quality of tears have improved significantly following topical cAD-MSCs treatment based on Schirmers tear test-1 and tear break-up time. More than half of all patients were found improved in the tear quantity. In particular, 56.5% of the patients that were unresponsive to prior immunosuppressant therapy had an effective increase in tear volume. The severity of clinical signs was also ameliorated according to the numeric rating scale score from both patient owners and the clinician. CONCLUSION: To sum up, topical cAD-MSCs may be beneficial especially in KCS patients with poor owner compliance for frequent daily use of eye drops or those who are unresponsive to immunosuppressant therapy.

In Vivo Confocal Microscopic Study of Hard Contact Lens-Induced Lipid Keratopathy Secondary to Corneal Neovascularization in a Rabbit Hypercholesterolemic Model.

OBJECTIVES: In vivo confocal microscopy was used to observe the morphological presentations and anatomical correlations between corneal neovascularization (NV) and intracorneal lipid deposition in a rabbit model of contact lens (CL)-induced lipid keratopathy secondary to corneal NV. METHODS: Rabbits were divided into 3 groups: (1) 8-week normal diet, (2) 8-week high-cholesterol diet, and (3) 4-week normal diet followed by 4-week high-cholesterol diet. Corneal NV was induced by closed-eye CL. The formation and maturation of corneal NV were shown by immunohistochemical staining against CD31 and high-molecular-weight melanoma-associated antigen. In vivo confocal microscopy identified corneal NV and lipid deposition. Acquired images for each eye were arranged and mapped into subconfluent montages. RESULTS: In group 1, corneal NV sprouting formed from the peripheral to the central cornea by the end of week 4. Pericytes around vessels were shown after 2 weeks of CL wear. In group 2, lipid deposition started from the peripheral cornea and progressively covered the whole cornea. In group 3, lipid deposition was found first in the central cornea after 2 weeks of high-cholesterol diet and progressed to cover the peripheral cornea. In vivo confocal microscopy demonstrated four different patterns of intracorneal lipid deposition: spindle shapes arranged randomly or in parallel, amorphous shapes, multiangular shapes, and mixed types. Intracorneal lipid deposition was distributed from basal corneal epithelium to deep stroma. CONCLUSIONS: Intracorneal lipids tend to accumulate around newly formed corneal NV but can extend to the area covered with mature NV. In vivo confocal microscopy can demonstrate various shapes and depths of intracorneal lipid deposition.

Glucose Transporter Type 4 Redistribution on the Membrane Induced by Insulin through Akt in Hydrocortisone Treatment in Rat Skeletal Muscles.

Hydrocortisone is a growth hormone frequently used in the treatment of low back pain. Hydrocortisone treatment has an anti-inflammation effect, which also inactivates glucose transporter type 4 (GLUT4) by p38 mitogen-activated protein kinase (MAPK) inhibition. Translocation of GLUT4 regulates body glucose homeostasis and muscle repair and is induced by insulin. In this study, 56 SD rats were divided into seven groups, and were treated with insulin or hydrocortisone in sedentary or exercise training groups. The muscle proteins and biochemical blood parameters were analyzed after 7 days of treatments. The results showed that the serum glucose increased in hydrocortisone treatment accompanied by GLUT4 inactivation in both the sedentary and exercise training rats. In the exercise training groups, GLUT4 was redistributed on the plasma membrane on co-treatment with insulin and hydrocortisone through Akt phosphorylation. Insulin treatment exerted a compensatory feedback effect on the GLUT4 translocation on hydrocortisone co-treatment, which was the cause of GLUT4 inactivation.

Characterization of protein marker expression, tumorigenicity, and doxorubicin chemoresistance in two new canine mammary tumor cell lines.

BACKGROUND: Canine mammary tumors (CMTs) are the most common type of cancer found in female dogs. Establishment and evaluation of tumor cell lines can facilitate investigations of the biological mechanisms of cancer. Different cell models are used to investigate genetic, epigenetic, and cellular pathways, cancer progression, and cancer therapeutics. Establishment of new cell models will greatly facilitate research in this field. In the present study, we established and characterized two new CMT cell lines derived from a single CMT. RESULTS: We established two cell lines from a single malignant CMT specimen: DTK-E and DTK-SME. Morphologically, the DTK-E cells were large, flat, and epithelial-like, whereas DTK-SME cells were round and epithelial-like. Doubling times were 24 h for DTK-E and 18 h for DTK-SME. On western blots, both cell lines expressed cytokeratin AE1, vimentin, cytokeratin 7 (CK7), and heat shock protein 27 (HSP27). Moreover, investigation of chemoresistance revealed that DTK-SME was more resistant to doxorubicin-induced apoptosis than DTK-E was. After xenotransplantation, both DTK-E and DTK-SME tumors appeared within 14 days, but the average size of DTK-SME tumors was greater than that of DTK-E tumors after 56 days. CONCLUSION: We established two new cell lines from a single CMT, which exhibit significant diversity in cell morphology, protein marker expression, tumorigenicity, and chemoresistance. The results of this study revealed that the DTK-SME cell line was more resistant to doxorubicin-induced apoptosis and exhibited higher tumorigenicity in vivo than the DTK-E cell line. We anticipate that the two novel CMT cell lines established in this study will be useful for investigating the tumorigenesis of mammary carcinomas and for screening anticancer drugs.

Evaluation of potential topical and systemic neuroprotective agents for ocular hypertension-induced retinal ischemia-reperfusion injury.

OBJECTIVE: To evaluate for drugs with superior neuroprotective efficacy and investigate their underlying mechanisms related to antioxidation. PROCEDURES: Brinzolamide (1%), timolol (0.5%), minocycline (22 mg/kg), lidocaine (1.5 mg/kg), and methylprednisolone (30 mg/kg) were administered to Sprague-Dawley (SD) rats. The retina was evaluated by electroretinography and histological analysis. The antioxidative capacity of drugs was evaluated to clarify the underlying mechanism. The oxidant/antioxidant profiles of plasma, red blood cells, and retina were analyzed by lipid peroxidation (malondialdehyde) and by measuring the activities of antioxidants. Proteomic analysis was used to investigate the possible protective mechanisms of the drug against ischemia-reperfusion injury. RESULTS: The results suggested that timolol, methylprednisolone, and minocycline protected retinal function. Methylprednisolone and minocycline possessed good antioxidative activity. Brinzolamide and lidocaine preserved the structural integrity of the retina, but not retinal function. CONCLUSION: Methylprednisolone, minocycline, and timolol have potential acute or delayed benefit in retinal ischemia-reperfusion injury. Their neuroprotective actions depend at least partially on the ability to alleviate oxidative stress.

Investigation of topical amniotic membrane suspension and ReGeneraTing Agent on early corneal stromal healing in rats.

Innovative topical bioregenerative materials promoting corneal stromal healing provide valuable alternatives for treating patients with deep corneal ulcers, and particularly beneficial for those with a higher anaesthetic risk. This study aimed to investigate the effects of topical amniotic membrane suspension (AMS) and ReGeneraTing Agent (RGTA) on surgically induced deep stromal ulcers in rats. Eighteen Sprague-Dawley rats were divided into 3 treatment groups: control group (topical normal saline, TID); AMS group (topical AMS, TID); RGTA group (topical RGTA, Q2D). Corneal microsurgery was used to create deep stromal ulcer. Evaluations were performed by corneal opacity grading, spectral domain optical coherence tomography (SD-OCT), histopathology, and immunohistochemistry. One-way ANOVA and Dunnett's test were used for statistical analysis. By the seventh day of treatment, both the AMS and RGTA groups showed significantly greater thickness in corneal stroma (both p-value < 0.05) than the control group. Additionally, the RGTA group exhibited a significantly higher degree of myofibroblast infiltration in the stroma and a greater level of corneal opacity (p < 0.05). No significant differences in the count of inflammatory cells were noted. In conclusion, both AMS and RGTA have demonstrated effectiveness in promoting the early stages of stromal wound healing and wound defect recovery in our research. Both AMS and RGTA have good potential for treating deep corneal ulcers in small animals practice. Further research is necessary to investigate the long-term effects and mechanism of using topical AMS and RGTA on treating deep corneal ulcer in clinical practice.

Characterizing the Robustness of Distinct Clinical Assessments in Identifying Dry Eye Condition of Animal Models.

PURPOSE: The study aims to characterize the robustness of distinct clinical assessments in identifying the underlying conditions of dry eye disease (DED), with a specific emphasis on the involvement of conjunctival goblet cells. METHODS: Seven rabbits receiving surgical removal of the lacrimal and Harderian glands were divided into two groups, one with ablation of conjunctival goblet cells by topical soaking of trichloroacetic acid (TCA) to the bulbar conjunctiva (n = 3) and one without (n = 4), and the conditions of DED were assessed weekly using Schirmer test, tear breakup time (TBUT), tear osmolarity, and National Eye Institute (NEI) fluorescein staining grading. After 8 weeks, the rabbits were sacrificed, and the eyes were enucleated for histopathological examination. RESULTS: Histopathological analysis revealed corneal epithelial thinning in both groups. While TCA soaking significantly decreased the density of conjunctival goblet cells, DED rabbits without TCA also showed a partial reduction in goblet cell density, potentially attributable to dacryoadenectomy. Both groups showed significant decreases in Schirmer test and TBUT, as well as an increase in tear osmolarity. In DED rabbits with TCA soaking, tear osmolarity increased markedly, suggesting that tear osmolarity is highly sensitive to loss and/or dysfunction of conjunctival goblet cells. Fluorescein staining was gradually and similarly increased in both groups, suggesting that fluorescein staining may not reveal an early disruption of the tear film until the prolonged progression of DED. CONCLUSION: The Schirmer test, TBUT, tear osmolarity, and NEI fluorescein grading are distinct, yet complementary, clinical assessments for the evaluation of DED. By performing these assessments in definitive DED rabbit models, both with and without ablation of conjunctival goblet cells, the role of these cells in the homeostasis of tear osmolarity is highlighted. Characterizing the robustness of these assessments in identifying the underlying conditions of DED will guide a more appropriate management for patients with DED.

Amygdala opioid receptors mediate the electroacupuncture-induced deterioration of sleep disruptions in epilepsy rats.

BACKGROUND: Clinical and experimental evidence demonstrates that sleep and epilepsy reciprocally affect each other. Previous studies indicated that epilepsy alters sleep homeostasis; in contrast, sleep disturbance deteriorates epilepsy. If a therapy possesses both epilepsy suppression and sleep improvement, it would be the priority choice for seizure control. Effects of acupuncture of Feng-Chi (GB20) acupoints on epilepsy suppression and insomnia treatment have been documented in the ancient Chinese literature, Lingshu Jing (Classic of the Miraculous Pivot). Therefore, this study was designed to investigate the effect of electroacupuncture (EA) stimulation of bilateral Feng-Chi acupoints on sleep disruptions in rats with focal epilepsy. RESULTS: Our result indicates that administration of pilocarpine into the left central nucleus of amygdala (CeA) induced focal epilepsy and decreased both rapid eye movement (REM) sleep and non-REM (NREM) sleep. High-frequency (100 Hz) EA stimulation of bilateral Feng-Chi acupoints, in which a 30-min EA stimulation was performed before the dark period of the light:dark cycle in three consecutive days, further deteriorated pilocarpine-induced sleep disruptions. The EA-induced exacerbation of sleep disruption was blocked by microinjection of naloxone, µ- (naloxonazine), κ- (nor-binaltorphimine) or δ-receptor antagonists (natrindole) into the CeA, suggesting the involvement of amygdaloid opioid receptors. CONCLUSION: The present study suggests that high-frequency (100 Hz) EA stimulation of bilateral Feng-Chi acupoints exhibits no benefit in improving pilocarpine-induced sleep disruptions; in contrast, EA further deteriorated sleep disturbances. Opioid receptors in the CeA mediated EA-induced exacerbation of sleep disruptions in epileptic rats.

Activation of amygdala opioid receptors by electroacupuncture of Feng-Chi (GB20) acupoints exacerbates focal epilepsy.

BACKGROUND: The effect of seizure suppression by acupuncture of Feng-Chi (GB20) acupoints has been documented in the ancient Chinese literature, Lingshu Jing (Classic of the Miraculous Pivot), however, there is a lack of scientific evidence to prove it. This current study was designed to elucidate the effect of electroacupuncture (EA) stimulation of bilateral Feng-Chi (GB20) acupoints on the epileptic activity by employing an animal model of focal epilepsy. METHODS: Administration of pilocarpine into the left central nucleus of amygdala (CeA) induced the focal epilepsy in rats. Rats received a 30-min 100 Hz EA stimulation of bilateral Feng-Chi acupoints per day, beginning at 30 minutes before the dark period and performing in three consecutive days. The broad-spectrum opioid receptor antagonist (naloxone), µ-receptor antagonist (naloxonazine), δ-receptor antagonist (naltrindole) and κ-receptor antagonist (nor-binaltorphimine) were administered directly into the CeA to elucidate the involvement of CeA opioid receptors in the EA effect. RESULTS: High-frequency (100 Hz) EA stimulation of bilateral Feng-Chi acupoints did not suppress the pilocarpine-induced epileptiform electroencephalograms (EEGs), whereas it further increased the duration of epileptiform EEGs. We also observed that epilepsy occurred while 100 Hz EA stimulation of Feng-Chi acupoints was delivered into naïve rats. EA-induced augmentation of epileptic activity was blocked by microinjection of naloxone, µ- (naloxonazine), κ- (nor-binaltorphimine) or δ-receptor antagonists (natrindole) into the CeA, suggesting that activation of opioid receptors in the CeA mediates EA-exacerbated epilepsy. CONCLUSIONS: The present study suggests that high-frequency (100 Hz) EA stimulation of bilateral Feng-Chi acupoints has no effect to protect against pilocarpine-induced focal epilepsy; in contrast, EA further exacerbated focal epilepsy induced by pilocarpine. Opioid receptors in the CeA mediated EA-induced exacerbation of focal epilepsy.

The protective effects of Lycium barbarum and Chrysanthemum morifolum on diabetic retinopathies in rats.

The effects of Lycium barbarum and Chrysanthemum morifolum extracts on diabetic retinopathy were evaluated. The diabetes model was induced by streptozotocin. Animals were divided into six groups: the control group received only vehicle; diabetic animal models received no treatment, insulin treatment, Lycium extract, Chrysanthemum extract, or a combination of Lycium and Chrysanthemum extracts, respectively. Retinal function was evaluated by electroretinography, and the diabetic progression was monitored by blood test for hyperglycemia. In addition, retinal histopathology and retinal glial reactivity were also investigated. The electroretinographic amplitudes of the a- and b-waves were significantly decreased in the diabetic animals, and the implicit time of the b-wave was also delayed, compared to the control group. However, reductions in the a- and b-wave amplitudes were not observed in the Lycium-treated group. Histopathological studies showed no significant differences between the Lycium-treated, Chrysanthemum-treated, Lycium/Chrysanthemum-treated groups, and the control group. The results of this study suggest that L. barbarum may have protective effects in diabetic retinopathy.

Features of ophthalmic, magnetic resonance imaging, and histopathology of a feline case of idiopathic sclerosing orbital pseudotumor.

In the present report, we describe a case of sclerosing orbital pseudotumor in an 11-year-old castrated male American Shorthair cat. Ophthalmic exam showed lagophthalmos, retracted right upper eyelid, and resistant to retropulsion in his right eye. Under magnetic resonance imaging (MRI) scans, increased volume of the extraocular muscles (EOMs) of the right eye was prominent. Immunosuppressive dosage of prednisolone partially ameliorated the clinical signs, but some clinical signs were still gradually progressive or persistent. In the second MRI scan, decreased diameter of the thickened right extraocular muscles was found. After the third MRI scan, enucleation of the right eye was performed due to substantial adverse effects of systemic steroid therapy. Histopathological examination revealed no evidence of neoplastic transformation nor infection. Feline restrictive orbital myofibroblastic sarcoma (FROMS) was therefore excluded, suggesting unknown causes of extensive fibrotic changes in the right orbit of the affected cat.

Animal models of dry eye: Their strengths and limitations for studying human dry eye disease.

Dry eye disease (DED), also called the keratoconjunctivitis sicca, is one of the most common diseases in the ophthalmology clinics. While DED is not a life-threatening disease, life quality may be substantially affected by the discomfort and the complications of poor vision. As such, a large number of studies have made contributions to the investigation of the DED pathogenesis and novel treatments. DED is a multifactorial disease featured with various phenotypic consequences; therefore, animal models are valuable tools suitable for the related studies. Accordingly, selection of the animal model to recapitulate the clinical presentation of interest is important for appropriately addressing the research objective. To this end, we systemically reviewed different murine and rabbit models of DED, which are categorized into the quantitative (aqueous-deficient) type and the qualitative (evaporative) type, based on the schemes to establish. The clinical manifestations of dry eye on animal models can be induced by mechanical or surgical approaches, iatrogenic immune response, topical eye drops, blockage of neural pathway, or others. Although these models have shown promising results, each has its own limitation and cannot fully reproduce the pathophysiological mechanisms that occur in patients. Nonetheless, the animal models remain the best approximation of human DED and represent the valuable tool for the DED studies.

Feline hypertrophic osteopathy: a collection of seven cases in Taiwan.

Between October 2003 and May 2004, seven cats were diagnosed with severe and extensive hypertrophic osteopathy of the appendicular skeleton without detectable underlying causes. All cats showed similar clinical signs of pain with progressive lameness of the limbs. One cat died shortly after presentation, whereas conditions of the others resolved after medical treatment and a change in diet. Regression of the bone lesions was observed radiographically in all surviving six cases.

The neuroprotective effect of submicron and blended Lycium barbarum for experiment retinal ischemia and reperfusion injury in rats.

The purpose of this study was to investigate the neuroprotective potential of submicron (milled) and blended Lycium barbarum (LB) in glaucomatous retinal neuropathy using a rat model of high intraocular pressure (HIOP) induced retinal ischemia. The rats were treated with 500, 250, 100 mg/kg LB (submicron or blended form) orally once daily for 56 days respectively after 1 week of retinal ischemia induction. We conducted electroretinography (ERG), histopathological analysis in retina and antioxidative level assays, such as total glutathione (GSH (glutathione) + reduced glutathione) + GSSH (glutathione disulfide), catalase activity, SOD (superoxide dismutase) activity, and lipid peroxidant malondialdehyde (MDA) in the retina and plasma of test rats. The results indicated that the amplitudes of a and b wave of ERG were preserved in rats treated with submicron and blended LB groups, the best protective effect on ERG b wave amplitudes was observed at the dosage of 250 mg/kg of both forms of LB. Retinal thickness was best preserved, particularly significant in the retinal inner nuclear layer in submicron 250 mg/kg LB group. The levels of antioxidant GSSH+GSH, SOD and catalase activity in the retina were higher in blended 500 mg/kg and submicron 250 mg/kg groups than other groups, while the MDA level was lower in submicron LB groups than that in blended LB and non-LB IR group. In the plasma, there was no significant difference in the levels of GSSH+GSH and catalase activity between treated groups, but higher levels of SOD and lower levels of MDA were observed in 250 mg/kg submicron and 500 mg/kg submicron LB groups than the blended LB and non-LB IR groups. Generally better antioxidative effects were observed in the submicron LB than blended LB among treated groups, especially the 250 mg/kg submicron LB, providing good retinal neuroprotection by preserving retinal structure and function with improved antioxidative capacity. The submicron LB may have clinical implication as an adjuvant therapy of oxidative stress and retinal damage caused by HIOP induced retinal ischemia and reperfusion injury.

In vivo confocal microscopic findings of corneal wound healing after corneal epithelial debridement in diabetic vitrectomy.

PURPOSE: To study healing of corneal wounds using in vivo confocal microscopy in patients who received corneal epithelial debridement during pars plana vitrectomy for proliferative diabetic retinopathy and to investigate risk factors for delayed healing. DESIGN: Prospective, observational case series. PARTICIPANTS: Forty-four eyes of 40 patients were enrolled. METHODS: In vivo confocal microscopy was used to evaluate selected images of the corneal basal and apical surface epithelial cells and subbasal nerves before surgery, weekly for the first month, and at 3 and 6 months after surgery. Slit-lamp biomicroscopy was carried out at the same time. Multiple linear regression analysis of selected potential risk factors was performed to investigate the main determinants of delayed corneal healing. MAIN OUTCOME MEASURES: Healing rate of corneal epithelial cells and subbasal nerves and factors influencing the healing. RESULTS: By slit-lamp biomicroscopy, corneal epithelial defects were found in 22.8% of eyes at 2 weeks and in 5.4% at 1 month after surgery. In vivo confocal microscopy demonstrated incomplete healing of basal epithelial cells in 72.1%, 15.2%, and 0% of eyes and incomplete healing of surface apical epithelial cells in 81.1%, 9.1%, and 0% of eyes at 1, 3, and 6 months after surgery. The percentage of subbasal nerves regaining preoperative appearance was 0%, 6.8%, and 89.3% at 1, 3, and 6 months after surgery. Regression analysis revealed infusion of silicone oil (P = 0.020) and C(3)F(8) (P = 0.017) resulted in delayed healing by slit-lamp biomicroscopy; age (P = 0.028), diabetic treatment regimen (P = 0.014), and scleral buckling (P = 0.001) correlated with delayed recovery of basal cells by in vivo confocal microscopy. The latter 2 factors also were related to delayed reconformation of apical cells (P = 0.011 and 0.004, respectively). Neither healing of apical and basal cells showed a significant correlation to findings by slit-lamp biomicroscopy (r = 0.19 and 0.09). CONCLUSIONS: Healing of corneal epithelial wounds in diabetic eyes is slow. Both the basal and apical epithelial layers were involved in the slow healing process. Age, diabetic treatment regimen, and several intraoperative factors may alter healing rates. FINANCIAL DISCLOSURE(S): The author(s) have no proprietary or commercial interest in any materials discussed in this article.

A 12-year retrospective study of canine testicular tumors.

To provide up-to-date information on the incidence and risk factors of canine testicular tumors, 476 pathological reports of male canine tumors from two institutes of veterinary medicine in Taiwan over a period of 12 years were reviewed. The detection rate of testicular tumors was 16.8% (80/476) in male canine tumors, and 94.1% (80/85) in male genital tumors. Ninety-six testicular tumors from 80 dogs were identified in this study, including 33 (34.4%) seminomas (SEMs), 25 (26%) interstitial cell tumors (ICTs), 22 (22.9%) mixed germ cell-stromal cell tumors (MGCSCTs), and 16 (16.6%) Sertoli cell tumors (SCTs). Of the 96 testicular tumors, 52 (54.2%) tumors developed from cryptorchid testes in 45 dogs (56.3%), and 44 (45.8%) tumors developed from scrotal testes in 35 dogs (44.7%). Cryptorchidism was significantly associated with development of MGCSCTs, SCTs, and SEMs, but not ICTs (p<0.01). The detection rate of testicular tumors in the dog younger than 10-years-old was significantly associated with cryptorchidism (p<0.01). Except for mixed breed dogs, the Maltese breed had high detection rate of testicular tumors, and may have high risk in cryptorchidism in this study. In conclusion, our results indicate that cryptorchidism alone or in addition to age will significantly affect the incidence and type of canine testicular tumors, and a high detection rate of testicular tumors in Maltese dogs is documented for the first time.

The investigation of histopathology and locations of excised eyelid masses in dogs.

BACKGROUND: Eyelid masses are the most common ocular diseases in dogs. However, there are no studies specifically investigating the location of eyelid masses. METHODS: 118 dogs with 119 eyelid masses were included. Medical records and pathology reports were retrospectively reviewed at National Taiwan University veterinary hospital and Vision Eyecare Centre for Animals between 2012 and 2017. RESULTS: Mean age of dogs was 9.4±2.5 years. Female spayed dogs had significantly higher prevalence of eyelid mass than female intact dogs (p<0.01). Prevalence of upper eyelid mass was significantly higher than lower eyelid mass (p<0.01). The upper lateral eyelid was the significantly more common location compared with the upper medial eyelid. Mean mass volume was 258.2±661.0 mm3. The most common eyelid mass type was meibomian epithelioma (34.5%), followed by meibomian adenoma (29.4%) and meibomian hyperplasia (18.5%). Non-tumours comprised 25.2%, benign tumours comprised 67.2% and malignant tumours comprised 7.6% of all eyelid masses. Malignant tumours were significantly larger than benign lesions (p<0.01). All dogs underwent surgical excision without any complications. CONCLUSION: The majority of eyelid masses were benign. Surgical intervention can prevent further ocular irritation and provide good prognosis.

Modulation of immediate early gene expression by tristetraprolin in the differentiation of 3T3-L1 cells.

Tristetraprolin (TTP) is a zinc-finger-containing AU-rich elements (ARE)-binding protein. AREs presented in the 3'untranslated region (UTR) of mRNAs from many proto-oncogenes, cytokines, and growth factors may be targets for regulation of messenger RNA stability. In this study, we observed that many immediate early genes (IEGs) were induced during the early differentiation of 3T3-L1 preadipocytes and their ARE-containing transcripts were degraded rapidly. Immunoprecipitation followed by RT-PCR analysis showed that two of IEG mRNAs, COX-2 (cyclooxygenase-2) and MKP-1 (mitogen-activated protein kinase phosphatase), were the target of TTP. Biotinylated MKP-1 AREs also could bring down TTP and the other ARE-binding protein HuR. RNA EMSA and competition assays showed that each of three AREs located in 3'UTR of MKP-1 mRNA has differential binding affinity to TTP. Sequence analysis of 3'UTR of IEG mRNAs suggested that TTP may prefer binding to UUAUUUAUU sequence. Taken together, our results implied that TTP may target specific ARE-containing IEGs' mRNAs such as COX-2 and MKP-1 mRNAs to modulate their expression post-transcriptionally.

Maxillary bone epithelial cyst in an adult miniature schnauzer.

Maxillary bone epithelial cyst is rare in dogs. A 5-year-old, spayed female miniature schnauzer developed a swelling below the nasal canthus of left eye. Plain radiograph demonstrated a 1.5 cm diameter of radiolucent lesion on the maxillary bone anteroventral to the eye, and contrast dacryocystorhinography confirmed an obstructed nasolarcrimal duct. The swelling showed poor response to antibiotic treatment but responded well to oral prednisolone. Exploratory surgery revealed a cyst-like structure filled with brown serous fluid. Histopathological examination of the removed cyst revealed a double cuboidal epithelial cyst. The dog recovered rapidly after surgery, and the swelling had not recurred for a 36-month follow-up. It is the first case of periorbital bone epithelial cyst reported in an adult miniature schnauzer.

The Neuroprotective and antioxidative effects of submicron and blended Lycium barbarum in experimental retinal degeneration in rats.

The object is to determine the neuroprotective and antioxidative effects of submicron and blended Lycium barbarum (LB) on retinal degeneration as evaluated by ERG, retinal histopathology and assays of antioxidant (total GSH) and peroxidant (MDA) in the retina. A rat model of light-induced retinal degeneration was used to assess the protective effect of different forms of Lycium barbarum (LB) on retinal degeneration. Rats were divided into four experimental groups, normal control, light-induced untreated, submicron LB and blended LB treated. The rats of submicron and blended groups were treated with 250 mg/kg LB orally once daily for 54 days, followed by induction of retinal degeneration. Retinal function was assessed by electroretinography (ERG). Enzyme-linked immunosorbent assay of the retina lysates was measured for the levels of antioxidants, reduced glutathione and glutathione disulfide, and peroxidants, malondialdehyde, in the retina. The ERG results showed a protective effect in LB treated groups with a greater effect observed in submicron LB treated group than the blended LB treated group. There were higher levels of GSH plus GSSG and lower MDA in submicron LB treated group than other groups. In conclusion, LB provided protective and antioxidative effects on the rat retina with light-induced retinal degeneration. Submicron LB protected degenerative retina better than blended LB. LB is effective against oxidative stress in the degenerative retina.