Anti-VEGF agents were found to have clinical implications for the successful treatment of vascular-driven diseases in humans. In this study, a detailed biological characterization of bevacizumab in a ...variety of in vitro assays was carried out to determine the effect of bevacizumab on equine umbilical vein endothelial cells (EqUVEC). EqUVECs were harvested from umbilical cords of clinically healthy horses and exposed to different concentrations (1, 2, 4, 6, 8 mg/mL) of bevacizumab (Avastin®). Assays concerning the drug’s safety (cell viability and proliferation assay) and efficacy (cell tube formation assay, cell migration assay, and Vascular endothelial growth factor (VEGF) expression) were carried out reflecting multiple cellular processes. Bevacizumab significantly decreased VEGF expression at all concentrations over a 72 h period. No cytotoxic effect of bevacizumab on EqUVECs was observed at concentrations of 4 mg/mL bevacizumab or lower. Incubated endothelial cells showed delayed tube formation and bevacizumab efficiently inhibited cell migration in a dose-dependent manner. Bevacizumab potently inhibits VEGF-induced cellular processes and could be a promising therapeutic approach in vascular-driven diseases in horses.
Squamous cell carcinoma (SCC) seriously compromises the health and welfare of affected horses. Although robust evidence points to equine papillomavirus type 2 (EcPV2) causing genital lesions, the ...etiopathogenesis of equine SCC is still poorly understood. We screened a series of SCCs from the head-and-neck (HN), (peri-)ocular and genital region, and site-matched controls for the presence of EcPV2-5 and herpesvirus DNA using type-specific EcPV PCR, and consensus nested herpesvirus PCR followed by sequencing. EcPV2 DNA was detected in 45.5% of HN lesions, 8.3% of (peri-)ocular SCCs, and 100% of genital tumors, whilst control samples from tumor-free horses except one tested EcPV-negative. Two HNSCCs harbored EcPV5, and an ocular lesion EcPV4 DNA. Herpesvirus DNA was detected in 63.6%, 66.6%, 47.2%, and 14.2% of horses with HN, ocular, penile, and vulvar SCCs, respectively, and mainly identified as equine herpesvirus 2 (EHV2), 5 (EHV5) or asinine herpesvirus 5 (AsHV5) DNA. In the tumor-free control group, 9.6% of oral secretions, 46.6% of ocular swabs, 47% of penile samples, and 14.2% of vaginal swabs scored positive for these herpesvirus types. This work further highlights the role of EcPV2 as an oncovirus and is the first to provide information on the prevalence of (gamma-)herpesviruses in equine SCCs.
To date the influence of herpesviruses on the development of equine ocular diseases has not been clearly determined.
The purpose of this study was to illustrate the course of equine ocular findings ...over a period of 18 months at 6 month intervals, in correlation with the results of herpesvirus detection.
266 Lipizzaners in 3 federal states of Austria underwent complete ophthalmologic examination 4 times. Blood samples, nasal- and conjunctival swabs were obtained at the same time and used for the detection of the equid gammaherpesviruses EHV-2 and EHV-5 using consensus herpesvirus PCR and type-specific qPCRs. Ophthalmic findings and results of herpesvirus PCRs were recorded and statistically analysed using one-way ANOVA, and multiple logistic regression analysis to determine the influence of herpesvirus infections and other contributing factors on the presence of ophthalmic findings.
In the first, second, third and fourth examination period 266, 261, 249 and 230 horses were included, respectively. Ophthalmic findings consistent with herpesvirus infections included conjunctival- and corneal pathologies. Statistical analysis revealed that the probability of positive herpesvirus PCR results decreased with progressing age; however the presence of corneal findings increased over time. At the time of each examination 45.1%, 41.8%, 43.0%, and 57.0% of horses with conjunctival or corneal findings, respectively, were positive for EHV-2 and/or EHV-5. However, 31.6%, 17.6%, 20.1%, and 13.0% of clinically sound horses were positive for these herpesviruses at each examination period, too.
Based on the results of our study there is a significant influence of young age on EHV-2 and/or EHV-5 infection. Corneal pathologies increased over time and with progressing age. Whether the identified findings were caused by herpesviruses could not be unequivocally determined.
Infections with
Encephalitozoon cuniculi in rabbits are observed at increasing frequency and are known as opportunistic infections in immunocompromised humans. 191 pet rabbits with suspected ...encephalitozoonosis, presented at the Animal Hospital of the Veterinary University of Vienna (Austria), were included in this study. Rabbits were serologically examined for antibodies against
E. cuniculi (144 positive out of 184 rabbits with suspected encephalitozoonosis compared to 14 positive out of 40 clinically healthy rabbits tested as part of a standard health check) and
Toxoplasma gondii (8 positive out of 157). Of the 144 seropositive rabbits with clinical signs, 75% showed neurological symptoms, 14.6% demonstrated phacoclastic uveitis and 3.5% suffered from renal failure. 6.9% of the animals had combined symptoms. Vestibular disease dominated within the rabbits that showed neurological symptoms. Polymerase chain reaction (PCR) could not detect parasite DNA in urine or cerebrospinal fluid (CSF), but did so in 4 out of 5 samples of liquefied lens material in cases with phacoclastic uveitis due to lens capsule rupture. Additionally further diagnostic procedures, such as inspection of the external ear canal (
N
=
69), radiography of the tympanic bullae (
N
=
65) were performed to rule out differential diagnosis. 54.2% of the patients exhibiting neurological symptoms recovered within a few days, while 87.5% of the rabbits suffering from renal failure died or had to be euthanized.
A 2-year-old female mixed-breed canine patient from Namibia presented originally with chronic uveitis. A serum antibody titer and a PCR test performed on the aqueous humor were positive for ...encephalitozoon cuniculi. The left eye showed an immature anterior focal cortical cataract in the periphery with suspected lens capsule rupture and signs of chronic uveitis. An incipient anterior focal cortical cataract was also perceivable in the patient's right eye. Despite local treatment as well as systemic administration of carprofen, prednisolone, and fenbendazol recurrent uveitis occurred. The patient then underwent bilateral extracapsular lensextraction via phacoemulsification. A PCR test of the lens material was positive for encephalitozoon cuniculi strain III. Recurring uveitis and secondary glaucoma 10 months post-op resulted in permanent blindness of the left eye. The patient then continued to receive local anti-inflammatory treatment. The last recheck examination of both eyes, 31 month post-op, revealed no signs of uveitis. This is the first case reported of a cataract in a canine patient caused by encephalitozoon cuniculi strain III.
Objective To determine the effect of various mydriatics (1% atropine, 1% cyclopentolate, 0.5% tropicamide, 10% phenylephrine) on intraocular pressure (IOP) and pupil size (PS) in normal cats.
...Animals studied The mydriatics were tested in 10 adult ophthalmoscopically normal European Domestic Short‐haired cats.
Procedure Single‐dose drug studies were divided into placebo (vehicle of phenylephrine), 10% phenylephrine, 0.5% tropicamide, 1% cyclopentolate and 1% atropine. After measurement of IOP and pupil size (PS) at 8 a.m. on the first day, one drop of the tested drug was applied to one randomly selected eye. The IOP and PS were measured for a minimum of 36 h until the pupil returned to pretest size.
Results Ten per cent phenylephrine had no significant effect on IOP, and the effect on the pupil size was minimal (≤ 1 mm difference). One per cent atropine showed a maximal mean difference of 4.3 mmHg ± 4.1 mmHg in IOP compared to the untreated eye. These differences were statistically significant at 1, 3, 6, 12 and 16 h post‐treatment. At 1, 1.5 and 2 h after unilateral application of 1% cyclopentolate, the IOP in the treated eye was significantly higher than the IOP in the untreated eye with a maximal mean difference of 4.1 mmHg ± 4.5 mmHg. With 0.5% tropicamide, IOP of the treated eye was significantly higher than IOP of the untreated eye at 1 and 1.5 h after treatment with a maximal mean difference of 3.5 mmHg ± 3.3 mmHg. One per cent atropine caused the greatest increase in IOP, followed by 1% cyclopentolate and 0.5% tropicamide. The dilating effect of atropine, cyclopentolate and tropicamide lasted longer than the effect on intraocular pressure.
Conclusions The parasympatholytics 1% atropine, 1% cyclopentolate and 0.5% tropicamide caused a statistically significant elevation of IOP and produced mydriasis in the treated eye in normal cats, while the sympathomimetic 10% phenylephrine had no significant effect on IOP or pupil size.
Objective Guinea pigs have a very low threshold of corneal sensitivity and at the same time nearly no reflex tearing compared to dogs, cats, and horses. The question arose whether there is a general ...correlation between corneal sensitivity and the quantity of reflex tearing.
Animals studied Totally 160 animals of 8 different species (20 animals per species) were investigated.
Procedures The corneal touch threshold (CTT) was measured with a Cochet–Bonnet esthesiometer. The palpebral fissure length (PFL) was measured with a calliper ruler. The Schirmer tear test (STT) was modified by adapting the width of the STT strip to the PFL of every species. For the STT II, 0.4% oxybuprocaine was applied.
Results Corneal touch threshold: Cows (1.67 g/mm2), horses (1.23 g/mm2), sheep (1.13 g/mm2), goats (1.44 g/mm2), dogs (2.16 g/mm2), and cats (1.33 g/mm2) show similar CTT values. In contrast, rabbits (6.21 g/mm2) and guinea pigs (7.75 g/mm2) show a significantly lower CTT. Tear Production Difference STT I − STT II: Rabbits have the greatest decline in tear production with 38.4%, followed by sheep (33.3%), dogs (31.1%), cats (24.7%), cows (23.7%), horses (18.0%), and goats (14.0%). Guinea pigs have no decline, but a slight increase of −16.0%. Correlation CTT and STT II − STT I Difference: Pearson’s correlation coefficient shows a small, but significant correlation. The coefficient of determination can only forecast a value with 7.1% certainty.
Conclusions The high variance and low reproducibility of results suggest that the measuring devices are inappropriate to assess the evaluated parameters. Therefore, no assured correlation between the corneal sensitivity and the quantity of reflex tearing could be found.
A 17-year-old Appaloosa mare was referred for evaluation of presumed refractory keratitis of the left eye. Gross examination revealed ocular discomfort and corneal neovascularization with a nasal ...focal opacification affecting approximately 40% of the corneal surface. On ophthalmic examination, extensive subepithelial to mid-stromal vascular branching accompanied by a homogeneous white, dense opacification, which affected up to 80% of the total corneal thickness, were apparent. Signs of concurrent uveitis were absent. Deep-stromal lamellar keratectomy with a conjunctival pedicle graft was performed under general anesthesia. Histopathology confirmed a poorly differentiated corneal stromal invasive squamous cell carcinoma (SI-SCC) with neoplastic cell extension to the surgical margins. Postoperatively, 4 topical mitomycin C 0.04% chemotherapy cycles combined with oral firocoxib therapy were initiated. Seven months after surgery, regrowth of the SI-SCC was clinically suspected. A total volume of 1 ml bevacizumab 2.5% was administered in the standing sedated horse via 3 mid-stromal corneal injections. Four weeks later, intrastromal bevacizumab injections (ISBIs) were repeated, however, this time the solution was injected directly into the main corneal vessel branches.Seven weeks after the second ISBIs, the left eye was comfortable and significant remission of corneal vascularization and opacity was recognized. No recurrence has been noted for a follow-up period of more than 53 months.Equine SI-SCC usually has a very poor prognosis for globe maintenance. To the authors' knowledge this is the first report of well-tolerated intrastromal antivascular endothelial growth factor adjunctive therapy with bevazicumab 2.5% and SI-SCC resolution after a multimodal treatment approach.
Objective
Establishing an immunohistochemical approach for semi‐quantitative assessment of the presence of immunoglobulin G (IgG) in equine, canine, and feline corneas.
Procedures
Healthy corneas of ...horses, dogs, and cats, euthanized because of a fatal disease or an unrecoverable trauma unrelated to and without a history of ophthalmic disease were formalin‐fixed, paraffin‐embedded, and determined to be pathomorphologically healthy by light microscopy. Automated immunohistochemistry was performed using primary antibodies against IgG, biotin‐conjugated secondary antibodies, and streptavidin‐peroxidase, as well as diaminobenzidine for visualization. After counterstaining with hematoxylin, epithelium, stroma, Descemet´s membrane (DM), and endothelium were semi‐quantitatively scored for the presence of IgG on a 4‐grade scale (0 = no, 1 = faint, 2 = medium, 3 = strong staining) by light microscopy.
Results
Corneal specimens of 20 horses (40 eyes) with a median age of 15.5 years (range 2–31 years), 12 dogs (21 eyes) with a median age of 10.0 years (range 4–16), and 13 cats (24 eyes) with a median age of 10.0 years (range 2–18) were included in the study. Different sexes and breeds were represented. In all corneas (100%), significant medium signal intensity in the stroma was observed. Variable immunosignal was obtained in epithelium, DM, and endothelium.
Conclusion
This method reproducibly allows for the detection of IgG in healthy equine, canine, and feline corneas, particularly stroma. Semi‐quantitative results evidence medium presence of IgG in the corneal stroma. Further research is needed to evaluate IgG presence in diseased corneas.
This case study focused on the impact to DACA participants in a mid-western city enrolled at an urban school setting in a region where 30% of all residing immigrants are unauthorized (Capps & Ruiz ...Soto, 2016). The investigation aimed to understand if the language used during the 2016 election cycle altered trauma-related behaviors in the DACA population. The data collected during the study suggested that students who identified with the DACA group exhibited trauma-related behaviors different from behaviors previously observed, and the new behaviors were a result of election cycle rhetoric. Implications for counselor training were significant.
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