Paulo Sérgio de Moraes Barros

CORNEAL ENDOTHELIUM OF THE MAGELLANIC PENGUIN (SPHENISCUS MAGELLANICUS) BY SCANNING ELECTRON MICROSCOPY

Abstract The corneal endothelium is essential for the maintenance of the corneal transparency. The aim of this study was to examine the morphology of the endothelial surface and perform morphometric analysis of the normal corneal endothelial cells of the Magellanic penguin (Spheniscus magellanicus) using scanning electron microscopy. The present work demonstrates that the corneal endothelium of the Magellanic penguin is similar to those described in other vertebrates.

Morphological analysis of the corneal endothelium in eyes of dogs using specular microscopy

Both healthy eyes of 10 six-year-old male and female mongrel dogs were studied. With a contact specular microscope the corneal endothelium was examined. Endothelial cells were analyzed in the central and peripheral cornea. Morphological analysis with regard to polymegathism and pleomorphism was performed. Three images of each region with at least 100 cells were obtained. The analysis showed that polygonal cells formed a mosaic-like pattern uniform in size and shape. The predominant number of cells was hexagonal. The polymegathism index was 0.22. The study demonstrates that the morphology of the normal corneal endothelial cells of dogs is similar to that found in the human cornea.

Ultrasonographic aspects and biometry of Striped owl’s eyes (Rhinoptynx clamator)

OBJECTIVE To report the biometric values and ultrasonographic aspects of the normal eye of the Striped owl (Rhinoptynx clamator). Sample population  Twenty-seven healthy, free-living, adult Striped owls from the Ecological Park of Tiete Veterinary Ambulatory (Sao Paulo, Brazil). PROCEDURES Both eyes of all owls underwent B-mode ultrasonographic examination and biometry was performed for lens axial length (WL), depth of the anterior (AC) and vitreous (VC) chambers, axial length of the globe (LB) and the pecten oculi (LP) of both eyes, using a 12 MHz probe. The owls were manually restrained without sedation and the eyes were topically anesthetized. RESULTS Biometric and statistical findings were as follows: in the left eye, the means and standard deviations were: LB = 23.76 ± 0.92 mm, WL = 7.79 ± 0.27 mm, AC = 4.27 ± 0.47 mm, VC = 11.36 ± 0.29 mm and LP = 5.69 ± 0.50 mm; in the right eye, the values were: LB = 24.25 ± 0.79 mm, WL = 8.03 ± 0.40 mm, AC = 4.56 ± 0.52 mm, VC = 11.40 ± 0.25 mm, and LP = 5.68 ± 0.41 mm. No significant differences were found between left and right eyes measurements of LB, WL, AC, VC, and LP dimensions. CONCLUSIONS Ocular ultrasound aspects and biometric values of the Striped owl are reported. The studys results provide means for various ocular measurements. The ultrasound is an easy and safe exam to be performed in the Striped owls eyes.

Dose-dependent in vitro inhibition of rabbit corneal matrix metalloproteinases by an extract of Pothomorphe umbellata after alkali injury

The in vitro ability of Pothomorphe umbellata ethanolic crude extract to inhibit matrix metalloproteinase (MMP) in normal cornea and in cornea after alkali injury was demonstrated. Corneas of albino rabbits were injured with 1 N NaOH for 20 s. After 48 h the corneas were excised, homogenized and analyzed for MMP-9 (92 kDa), pro-MMP-2 (72 kDa) and MMP-2 (67 kDa) activity by gelatin zymography. The activity was also measured in untreated corneas. After electrophoresis of 20 microg protein, gels were incubated with 50, 100, or 250 microg/mL lyophilized hydroethanolic (1:1) root crude extract of P. umbellata standardized for 4-nerolidylcatechol (7.09%). The activity of the enzymes was compared with that of untreated gel. At 48 h after injury, the activity of all MMPs was increased compared with untreated eyes. When the gels were incubated with P. umbellata extract the activity of MMP-2, pro-MMP-2 and MMP-9 decreased in a dose-dependent manner. MMP-9 activity decreased by approximately 50% after incubation with 50 microg/mL and was completely abolished at 100 and 250 microg/mL of the extract. After incubation with 50 microg/mL the activity of pro-MMP-2 and MMP-2 also decreased by 50%. The activity of pro-MMP-2 was almost completely abolished after incubation with 250 microg/mL of the extract. For MMP-2 the incubation with 100 or 250 microg/mL of the extract of P. umbellata promoted a 10-fold decrease in activity. In conclusion, P. umbellata root crude extract can be useful as an alternative therapy to control MMP activity after corneal injury.

Criptococose canina: relato de caso

A case of cryptococcosis with ocular and cutaneous involvement is reported by the first time in Brazil in a dog. A two-year-old female German Shepherd living in a hold house with other five healthy dogs was infected through the contact with pigeon (Columba livia) feces. The illness started 90 days previously with cutaneous and bone involvement resulting in characteristic clinical signs and lesions, in addition to a initially unilateral asymptomatic chorioretinitis. The diagnosis was established based on anamnesis, physical and dermatologic examinations and complementary exams (radiographs, skin biopsy and histopathology) and the isolation of a Cryptococcus neoformans var. neoformans strain was accomplished. There was a complete resolution of the disease after nine months of therapy using itraconazole (9mg/kg, q24h, PO) and no side effect was observed.

Scanning electron microscopy of the corneal endothelium of ostrich

The aim of this study was to examine the endothelial surface morphology and perform a morphometric analysis of the corneal endothelial cells of ostrich (Struthio camelus) using scanning electron microscopy. Polygonality, mean cell area, cell density and coefficient of variation of mean cell area were analyzed. The normal corneal endothelium consisted of polygonal cells of uniform size and shape with few interdigitations of the cell borders. Microvilli appeared as protusions on the cellular surface. The average cell area was 269±18μm2 and the endothelial cell density was 3717±240cells mm -2 . The coefficient of variation of the cell area was 0.06, and the percentage of hexagonal cells was 75%. The parameters evaluated did not differ significantly between the right and the left eye from the same ostrich. The results of this study showed that the ostrich corneal endothelial cells appear quite similar to those of the other vertebrates.

Retrospective study of ocular disorders in Amazon parrots

A retrospective study was conducted to identify the occurrence and types of ocular disorders in 57 Amazon parrots admitted to the Ophthalmology Service, Veterinary Teaching Hospital, School of Veterinary Medicine, University of Sao Paulo, Brazil from 1997 to 2006. The most frequent observed disorder was cataracts, present in 24 of the 114 examined eyes (57 parrots). Uveitis, ulcerative keratitis and keratoconjunctivitis were frequently diagnosed as well. The cornea was the most affected ocular structure, with 28 reported disorders. Uveal disorders also were commonly observed. Conjunctiva and eyelid disorders were diagnosed in lower frequency. Results suggest that cataracts are common and that cornea, lens and uvea are the most affected ocular structures in Amazon parrots.

X-ray imaging in advanced studies of ophthalmic diseases

Microscopic characterization of pathological tissues has one major intrinsic limitation, the small sampling areas with respect to the extension of the tissues. Mapping possible changes on vast tissues and correlating them with large ensembles of clinical cases is not a feasible procedure for studying most diseases, as for instance vision loss related diseases and, in particular, the cataract. Although intraocular lens implants are successful treatments, cataract still is a leading public-health issue that grows in importance as the population increases and life expectancy is extended worldwide. In this work we have exploited the radiation-tissue interaction properties of hard x-rays--very low absorption and scattering--to map distinct lesions on entire eye lenses. At the used synchrotron x-ray photon energy of 20 keV (wavelength lambda=0.062 nm), scattering and refraction are angular resolved effects. It allows the employed x-ray image technique to efficiently characterize two types of lesions in eye lenses under cataractogenesis: distributions of tiny scattering centers and extended areas of fiber cell compaction. The data collection procedure is relatively fast; allowing dozens of samples to be totally imaged (scattering, refraction, and mass absorption images) in a single day of synchrotron beam time. More than 60 cases of canine cataract, not correlated to specific causes, were investigated in this first application of x-rays to image entire lenses. Cortical opacity cases, or partial opacity, could be related to the presence of calcificated tissues at the cortical areas, clearly visible in the images, whose elemental contents were verified by micro x-ray fluorescence as very rich in calcium. Calcificated tissues were also observed at nuclear areas in some cases of hypermature cataract. Total opacity cases without distinguishable amount of scattering centers consist in 70% of the analyzed cases, where remarkable fissure marks owing to extended areas of fiber cell compaction are diagnosed.

Antioxidant status of dog aqueous humor after extracapsular lens extraction

We determined the antioxidant status of the aqueous humor after extracapsular lens extraction in 14 mongrel dogs weighing about 10 kg. The animals were examined by slit lamp biomicroscopy, applanation tonometry and indirect ophthalmoscopy. One eye was submitted to conventional extracapsular lens extraction and the other was used as control. Samples of aqueous humor were obtained by anterior chamber paracentesis before and at days 1, 2, 3, 7 and 15 after surgery. Total antioxidant status was determined as the capacity of aqueous humor to inhibit free radical generation by 2,2-azobis(2-amidopropane) chlorine. Ascorbic acid concentration was measured by HPLC with UV detection. Protein content was determined with the biuret reagent. Statistical analysis was performed by ANOVA followed by the Tukey-Kramer test. Protein concentration increased from 0.61 to 22 mg/ml 24 h after surgery. These levels were maintained and returned to normal at day 7. Total antioxidant capacity was reduced from 50 to about 30 min until day 3 and at day 7 it was equal to control. Ascorbic acid levels were reduced from 252 to about 110 microM and then returned to control values at day 15. Considering the importance of ascorbic acid concentration in aqueous humor for the maintenance of the antioxidant status of the anterior segment of the eye, the decrease of antioxidant defenses suggests that the surgical procedures promote an oxidative stress condition in the eye.

Ceratoplastia lamelar em cães utilizando membrana fetal eqüina como enxerto: estudo experimental

Purpose: To evaluate the applicability of the equine fetal membrane (amniotic membrane, chorion and alantoid) as graft in lamelar keratoplasty in dogs. Methods: 9 mixed breed dogs were used, according to the ARVO statement for use of animals in ophthalmic and vision research. Superficial keratectomy was performed with a 5 mm trephine and a 6 mm diameter fragment of the fetal membrane that was sutured in place with 8-0 nylon simple interrupted stitches. Clinical examination were performed during 2, 7, 15 and 60 days and, afterwards, the animals were euthanized; the eyes were enucleated for histological study. Results: Clinically, one could observe slight corneal edema near the implant since the early phases, until the middle stage of the investigation. Neovascularization appeared progressively; its higher intensity was observed in the intermediate phase, disappearing gradually. At 60 days, one could notice a macula at the surgery site. The histological findings showed epithelialization and perfect integration of the graft to the receptor tissue at early stages; the cellular and vascular reactions were more intense in this phase. On day 15, the vascular elements were reduced in relation to the matrix and cellular elements. At any stage, inflammatory infiltrate in the graft, as well at the site of the suture were not observed. Conclusions: The equine fetal membrane (amniotic membrane, chorion and alantoid) can be useful as graft in lamellar keratoplasty in dogs.