In Situ Gelling Electrospun Ocular Films Sustain the Intraocular Pressure-Lowering Effect of Timolol Maleate: In Vitro, Ex Vivo, and Pharmacodynamic Assessment.

Most common intraocular pressure (IOP) reduction regimens for the management of glaucoma include the topical use of eye drops, a dosage form that is associated with short residence time at the site of action, increased dosing frequency, and reduced patient compliance. In situ gelling nanofiber films comprising poly(vinyl alcohol) and Poloxamer 407 were fabricated via electrospinning for the ocular delivery of timolol maleate (TM), aiming to sustain the IOP-lowering effect of the ß-blocker, compared to conventional eye drops. The electrospinning process was optimized, and the physicochemical properties of the developed formulations were thoroughly investigated. The fiber diameters of the drug-loaded films ranged between 123 and 145 nm and the drug content between 5.85 and 7.83% w/w. Total in vitro drug release from the ocular films was attained within 15 min following first-order kinetics, showing higher apparent permeability (Papp) values across porcine corneas compared to the drug's solution. The fabricated films did not induce any ocular irritation as evidenced by both the hen's egg test on chorioallantoic membrane and the in vivo Draize test. In vivo administration of the ocular films in rabbits induced a faster onset of action and a sustained IOP-lowering effect up to 24 h compared to TM solution, suggesting that the proposed ocular films are promising systems for the sustained topical delivery of TM.

Intraocular localization of Onchocerca lupi adult worm in a dog with anterior uveitis: A case report.

OBJECTIVE: Canine ocular onchocercosis may represent an important ocular disease with zoonotic potential and widespread geographical distribution. An uncommon localization of Onchocerca lupi adult worm in the anterior chamber of the eye is described for the first time. ANIMAL STUDIED: A 4-year-old, intact, male, mixed-breed dog was admitted with profuse lacrimation and severe squinting. On the right eye, there were mucopurulent discharge, chemosis, and all typical signs of anterior uveitis which were attributed to the presence of a moving filarial worm within the anterior chamber. PROCEDURE: Removal of the parasite from the anterior chamber by a limbal incision revealed a mature adult male, 7.65-cm-long O. lupi worm. Periocular skin scrapings for the corresponding microfilariae were negative. Postoperatively, apart from systemic prednisolone and doxycycline hyclate and a topical antibiotic/steroid solution, parenteral melarsomine, followed by ivermectin, was given until the complete disappearance of the ocular lesions. CONCLUSIONS: Intraocular onchocercosis should be included in the differential diagnosis of canine anterior uveitis in endemic areas. This case highlights the atypical localization of O. lupi in the canine eye emphasizing on the possibility for a complete cure after the surgical removal of the parasite and the filaricidal medication subsequently applied.

Mesenchymal stem cells improve healing of the cornea after alkali injury.

PURPOSE: To evaluate the efficacy of mesenchymal stem cells (MSCs) to ameliorate the consequences of corneal alkali injuries. METHODS: Corneal alkali injuries were created in 30 rabbit eyes. The MSC group (n = 15) were treated with intrastromal and subconjunctival injections of phosphate-buffered saline (PBS) containing 2 × 10(6) MSCs and topical application. The control group (n = 15) was treated with PBS by the same applications forms. Drops of standard treatment (ascorbate 10 %, citrate 10 %, tobramycin, dexamethasone, Cyclogyl) were instilled for 2 weeks. Rabbits underwent slit-lamp examination, fluorescein staining, photography, and were evaluated for corneal neovascularization, opacification, and epithelial defects. Tear secretion and IOP were also evaluated. Furthermore, the concentration of Serumglutamic-pyruvic transaminase (SGPT) and vascular endothelial factor (VEGF) were measured. Immunohistochemistry was also performed for a-SMA and Ki-67. RESULTS: Eyes treated with MSCs showed better recovery. The mean neovascularized area was significantly smaller in the MSC group (p < 0.05). A significant difference in the degree of corneal opacification and re-epithelialization was also observed, as well as the IOP at 21 and 28 posttraumatic days (p < 0.05). Histology showed that MSCs resulted in almost normal architecture of eye tissues. After the MSCs infusion, SGPT and VEGF levels in cornea were significantly reduced. Immunohistochemistry demonstrated a reduction of a-SMA in the MSC group with higher mitotic-regenerative activity with the presence of Ki67. CONCLUSIONS: Our study represents a first step in understanding the possibilities of the MSC approach to treatment of alkali injuries of the cornea and shows that such an approach improves clinical outcomes and leads to better prognosis.

Estimation of normal tear production in free-living Eurasian black vultures (Aegypius monachus) and griffon vultures (Gyps fulvus) in Dadia National Park, Greece.

The aim of this study was to record the Schirmer tear test I (STT I) measurements in free-living vultures in order to estimate normal values. The Eurasian black vulture (Aegypius monachus), which breeds in the Mediterranean region and Asia, is listed as near threatened; it is also classified as vulnerable at the European level and endangered in Greece. The griffon vulture (Gyps fulvus), once widespread across the continent, has undergone a dramatic decline which has led to its extinction in many regions. Sixty-two animals were examined in total including 54 black vultures and 8 griffon vultures. The birds were classified into five age groups while four age groups were then combined into one large group: free-flying. STT I measurements and complete ophthalmic examinations were performed. Mean STT I value for black vultures was 10.9 +/- 3.3 mm/min (right eye, oculus dexter, OD) and 11.9 +/- 3.3 mm/min (left eye, oculus sinister, OS) and for griffon vultures was 6.4 +/- 1.8 mm/min OD and 6.5 +/- 1.8 mm/min OS. In both eyes, STT I values in black vultures were significantly higher than those recorded in griffon vultures. Intraspecific comparisons yielded a significant difference between eyes of black vultures but not between those of griffon vultures, with OS producing higher STT I readings than did OD. When STT I was compared between OD and OS for each age group separately, a statistically significant difference was detected in the immature and free-flying black vultures. In addition, black vulture hatchlings had a significantly higher tear production than did free-flying juveniles, immatures, subadults, and adults. STT I values in black vultures are similar to those reported in other Accipitriformes but are lower in griffon vultures. This difference is probably related to anatomic, evolutionary, and feeding factors and requires further investigation.

Molecular characterization of a canine coronavirus NA/09 strain detected in a dog's organs.

In the present study, the detection of a pantropic canine coronavirus (CCoV) strain in a dog with lethal diarrhoea is reported. RT-PCR and real-time RT-PCR assays were used for the detection, characterization and quantitation of CCoV. Sequence and phylogenetic analysis of the CCoV NA/09 revealed a high degree of sequence identity with the pantropic strain CB/05, indicating the presence of CB/05-like pantropic strains in Greece. The absence of the 38-nucleotide deletion in ORF3b, which is characteristic of CB/05, indicates the need to identify new genetic markers for pantropic variants of CCoV, probably in the spike-protein gene region.

Effect of major versus minor mastectomy on host immunity in canine mammary cancer.

Surgical procedures can affect host immunity proportionally to the extent of surgical trauma. In cancer cases, surgery-induced immunosuppression can potentially promote tumour metastasis. The aim of this study was to investigate, in bitches with malignant mammary tumours, whether major surgery (total unilateral mastectomy or bilateral regional mastectomy) has a more negative effect than minor surgery (unilateral regional mastectomy) on components of host immunity. Twenty bitches with mammary cancer of clinical stage II or III were allocated to group A (minor surgery) or group B (major surgery) of 10 animals each receiving the same anaesthetic protocol for mastectomy. Immune cell measurements in blood [number of leukocytes, neutrophils, lymphocytes and platelets, and relative percentages of T-lymphocytes (CD3+) and their CD4+, CD8+ and CD5low+ subpopulations] were performed before anaesthesia (day 0) and on days 3 and 10 post-mastectomy. On day 3, leukocytes, neutrophils and platelets numbers were higher (p = 0.016, 0.032 and 0.017, respectively) in group B than in group A. For all 20 bitches, T-lymphocytes and the CD4+, CD5low+ T-cells were significantly decreased on day 3, but no significant differences were noted between groups. Minor mastectomy seemed to preserve innate immunity better than major mastectomy, but cellular immunity was rather equally affected.

Abortion Related to Toxoplasma gondii Infection in a Bactrian Camel (Camelus bactrianus) in Greece: A Case Report.

A female, 5 yr old Bactrian camel was presented to the Exotic and Wildlife Medicine Unit, School of Veterinary Medicine, Aristotle University of Thessaloniki, Thessaloniki, Greece, with severe dehydration, depression, anorexia, mild dyspnea and diarrhea. Supportive treatment immediately initiated with fluids, electrolytes and broad-spectrum antibiotics. The general condition of the animal was stable for the next 3 days, but at 4th day became worse, since the camel remained in sternal recumbency, denied to drink water and abortion of a mummified fetus was noticed. The aborted fetus and fetal membranes were submitted for laboratory examinations (bacterial cultures, MZN, cytology, PCR) that revealed Toxoplasma gondii infection. Treatment with sulfadimidine improved the situation of the animal that returned to its farm 1 week later. This seems to be the first reported case in the literature of confirmed toxoplasmic abortion in camels.

Evaluation of Clinical and Histological Outcomes of Adipose-Derived Mesenchymal Stem Cells in a Rabbit Corneal Alkali Burn Model.

To assess effects of adipose-derived mesenchymal stem cells (AMSCs) in corneal alkali injuries in an experimental animal model. Twenty white New Zealand rabbits were included in the study. The animal models were randomly divided into 2 groups. Rabbits in the AMSC group (n = 10) received an intrastromal, a subconjunctival injection, and topical instillation of 0.5 ml totally of phosphate-buffered saline (PBS) containing 2 × 106 AMSCs. In the control group (n = 10), rabbits received only 0.5 ml of PBS using the same methods. A masked investigator measured the corneal sensation, anterior chamber Inflammation (ACI), and conjunctival congestion. Additionally, a blind histological and immunohistochemical evaluation was made. In the AMSC group, the central corneal sensation was increased whereas ACI and conjunctival congestion were reduced compared to the control group in the 28 days of follow-up (p < 0.05). A statistically significant difference (p < 0.05) was noted between the two groups as recorded in the above parameters. Histological analysis showed that pathological vascularization was markedly reduced in the AMSC group which was consistent with the absence of factor VIII in the immunohistochemistry sections. There is a trend towards improved clinical outcomes including corneal sensation as well as acceleration in the restoration of normal corneal architecture in corneal alkali burns treated with AMSCs, results that support the need for further research in the field.