Birefringence analysis of collagen supraorganization in cat corneas with tropical keratopathy.

OBJECTIVE: To evaluate the birefringent properties of the cornea and examine the supraorganizational aspects of collagen fibers in cats with tropical keratopathy. PROCEDURE: In this study, 10-micrometer-thick sections of corneal tissue from cats with tropical keratopathy were examined, both in the opaque and transparent areas of the anterior stroma. Control samples were obtained from healthy cat corneas. Polarized light microscopy was employed to evaluate the birefringent properties using two distinct methods. The first method involved measuring the optical retardation associated with corneal birefringence, while the second method assessed the alignment/waviness of the birefringent collagen fibers. Differences were significant when p < .05. RESULTS: Tropical keratopathy resulted in a significant rise (p < .05) in optical retardation in both opaque and transparent regions of the cat cornea. In the anterior stroma, both the opaque zones and transparent tissue exhibited a higher degree of collagen fiber packing than the control corneas. However, no significant differences (p > .05) in alignment were observed between the transparent tissue of the diseased cornea and the healthy corneas. CONCLUSION: Supraorganizational changes in collagen fiber packing are not restricted to lesion zones in cat corneas affected by tropical keratopathy. Such alterations also occur in the corneal tissue of the anterior stroma adjoining the lesions. Therefore, it is plausible that the transparent tissue of the anterior stroma in corneas affected by the disease may have functional abnormalities, despite its macroscopic healthy appearance. Additional investigations are required to clarify the implications of these potential defects and their conceivable contribution to tropical keratopathy.

Accuracy of ARFI elastography in the differentiation of cataract stages in dogs

ABSTRACT: Objective was to evaluate the accuracy of elastography in the differentiation between normal and cataract lenses One hundred forty-five eyes of 98 dogs were divided into groups according to cataract stage. Forty-twoeyes were submitted to phacoemulsification. Biometric parameters, echogenicity and echotexture patterns of the anterior, posterior and vitreous chambers, lens and retina-choroid-sclera complexes were evaluated by ocular ultrasound in modes A and B. Deformability, and color (blue color = indicated less rigid structures, color red = more rigid structures) of the lenses were evaluated by the elastogram. The shear wave velocity (SWV; m/s) was calculated in three regions of the lens, both in the cortex and in the nucleus. The SWV of nucleus was statistically different between the normal lenses and with cataracts, and between the stages of cataract (P 0.001). Healthy lenses and incipient cataracts had a more rigid nucleus. Mature cataracts presented lowest nuclear rigidity (P 0.001). On cortical region the SWV was significantly higher (P 0.01) in intumescent and incipient cataracts. SWV less than 2.67m/s indicates cataract with a sensitivity of 72% and specificity of 94%. Values lower than 2.23m/s suggest mature cataract, with sensitivity of 71% and specificity of 76%. SWV greater than 2.66 m/s are associated with normal lenses or incipient cataract, presenting sensitivity of 94% and specificity of 84%. Qualitative method allowed differentiation between healthy and affected lenses and the classification of evolutionary stages. There was a correlation between the degree of stiffness of lens in cortical and nuclear regions (p=00165, r=0.37) and between the balanced saline solution quantitative and surgical time (P 0.01, r=0.73). Degree of stiffness of lens did not correlate with parameters of phacoemulsification. Elastographic proved feasible for evaluating the lens of dogs, characterizing the types of cataracts, and demonstrating increased stiffness of the diseased lenses.

RESUMO: O objetivo foi avaliar a precisão da elastografia na diferenciação entre lentes normais e de catarata. Cento e quarenta e cinco olhos de 98 cães foram divididos em grupos de acordo com o estágio de maturação da catarata. Quarenta e dois olhos foram submetidos à facoemulsificação. Parâmetros biométricos, ecogenicidade e padrões de ecotextura das câmaras anterior, posterior e vítrea, lente e complexos retina-coróide-esclera foram avaliados por ultrassonografia ocular nos modos A e B. A deformabilidade e a coloração (cor azul = indicou estruturas menos rígidas, cor vermelha = estruturas mais rígidas) das lentes foram avaliadas pelo elastograma. A velocidade da onda de cisalhamento (SWV; m/s) foi calculada em três regiões da lente, tanto no córtex quanto no núcleo. A SWV do núcleo foi estatisticamente diferente entre as lentes normais e com catarata e entre os estágios da catarata (P 0,001). Lentes saudáveis e cataratas incipientes tinham um núcleo mais rígido. Cataratas maduras apresentaram menor rigidez nuclear (P 0,001). Na região cortical, a SWV foi significativamente maior (P 0,01) nas cataratas intumescentes e incipientes. Uma SWV menor que 2,67m/s indica catarata com sensibilidade de 72% e especificidade de 94%. Valores inferiores a 2,23m/s sugerem catarata madura, com sensibilidade de 71% e especificidade de 76%. Uma SWV superior a 2,66m/s está associada à catarata normal ou incipiente, apresentando sensibilidade de 94% e especificidade de 84%. O método qualitativo permitiu a diferenciação entre lentes normais de olhos saudáveis e afetadas e a classificação dos estágios evolutivos. A elastografia se mostrara uma ferramenta viável para avaliar as lentes de cães, caracterizando os tipos de catarata e demonstrando maior rigidez das lentes doentes.

Accuracy of ARFI elastography in the differentiation of cataract stages in dogs / Acurácia da elastografia ARFI na diferenciação dos estágios da catarata em cães

Objective was to evaluate the accuracy of elastography in the differentiation between normal and cataract lenses One hundred forty-five eyes of 98 dogs were divided into groups according to cataract stage. Forty-twoeyes were submitted to phacoemulsification. Biometric parameters, echogenicity and echotexture patterns of the anterior, posterior and vitreous chambers, lens and retina-choroid-sclera complexes were evaluated by ocular ultrasound in modes A and B. Deformability, and color (blue color = indicated less rigid structures, color red = more rigid structures) of the lenses were evaluated by the elastogram. The shear wave velocity (SWV; m/s) was calculated in three regions of the lens, both in the cortex and in the nucleus. The SWV of nucleus was statistically different between the normal lenses and with cataracts, and between the stages of cataract (P<0.001). Healthy lenses and incipient cataracts had a more rigid nucleus. Mature cataracts presented lowest nuclear rigidity (P<0.001). On cortical region the SWV was significantly higher (P<0.01) in intumescent and incipient cataracts. SWV less than 2.67m/s indicates cataract with a sensitivity of 72% and specificity of 94%. Values lower than 2.23m/s suggest mature cataract, with sensitivity of 71% and specificity of 76%. SWV greater than 2.66 m/s are associated with normal lenses or incipient cataract, presenting sensitivity of 94% and specificity of 84%. Qualitative method allowed differentiation between healthy and affected lenses and the classification of evolutionary stages. There was a correlation between the degree of stiffness of lens in cortical and nuclear regions (p=00165, r=0.37) and between the balanced saline solution quantitative and surgical time (P<0.01, r=0.73). Degree of stiffness of lens did not correlate with parameters of phacoemulsification. Elastographic proved feasible for evaluating the lens of dogs, characterizing the types of cataracts, and demonstrating increased stiffness of the diseased lenses.(AU)

O objetivo foi avaliar a precisão da elastografia na diferenciação entre lentes normais e de catarata. Cento e quarenta e cinco olhos de 98 cães foram divididos em grupos de acordo com o estágio de maturação da catarata. Quarenta e dois olhos foram submetidos à facoemulsificação. Parâmetros biométricos, ecogenicidade e padrões de ecotextura das câmaras anterior, posterior e vítrea, lente e complexos retina-coróide-esclera foram avaliados por ultrassonografia ocular nos modos A e B. A deformabilidade e a coloração (cor azul = indicou estruturas menos rígidas, cor vermelha = estruturas mais rígidas) das lentes foram avaliadas pelo elastograma. A velocidade da onda de cisalhamento (SWV; m/s) foi calculada em três regiões da lente, tanto no córtex quanto no núcleo. A SWV do núcleo foi estatisticamente diferente entre as lentes normais e com catarata e entre os estágios da catarata (P<0,001). Lentes saudáveis e cataratas incipientes tinham um núcleo mais rígido. Cataratas maduras apresentaram menor rigidez nuclear (P<0,001). Na região cortical, a SWV foi significativamente maior (P<0,01) nas cataratas intumescentes e incipientes. Uma SWV menor que 2,67m/s indica catarata com sensibilidade de 72% e especificidade de 94%. Valores inferiores a 2,23m/s sugerem catarata madura, com sensibilidade de 71% e especificidade de 76%. Uma SWV superior a 2,66m/s está associada à catarata normal ou incipiente, apresentando sensibilidade de 94% e especificidade de 84%. O método qualitativo permitiu a diferenciação entre lentes normais de olhos saudáveis e afetadas e a classificação dos estágios evolutivos. A elastografia se mostrara uma ferramenta viável para avaliar as lentes de cães, caracterizando os tipos de catarata e demonstrando maior rigidez das lentes doentes.(AU)

THE EYE OF CRAB-EATING FOX (CERDOCYON THOUS): ANATOMICAL CHARACTERISTICS AND NORMATIVE VALUES OF SELECTED DIAGNOSTIC TESTS, MORPHOMETRY OF CORNEAL TISSUE, AND ARRANGEMENTS OF CORNEAL STROMAL COLLAGEN FIBERS.

This study aimed to evaluate the ophthalmic parameters, morphometric features of corneal tissue, and arrangements of corneal stromal collagen fibers in crab-eating fox (Cerdocyon thous), a species of neotropical wild canid. We conducted the study on six juvenile crab-eating foxes (12 eyes), whilst 16 eyes were obtained post mortem from eight adult crab-eating foxes. The research was divided into two stages. In the first stage, eye anatomical characteristics, tear production (Schirmer 1 tear test, STT1), intraocular pressure (IOP), ocular echobiometry, and specular microscopy parameters related to morphology of corneal endothelium were studied in juvenile animals. In the second stage, morphometric features of corneal tissue (central corneal thickness [CCT] and corneal epithelium thickness) and arrangements of stromal collagen fibers were studied using eyes from adult animals. The main findings were that crab-eating fox eyes have vertical-slit pupils, holangiotic retina, and reference values (mean ± SD) of 13.37 ± 3.79 mm/min for STT1 and of 10.43 ± 3.84 mmHg for IOP. The ocular echobiometric features observed in crab-eating foxes are different from those reported for domestic dogs (Canis familiaris). Conversely, the corneal endothelial parameters are similar to those of domestic dogs. The CCT measured by tissue morphometry was 0.54 ± 0.06 mm, and the corneal epithelium thickness was 60.13 ± 8.71 µm. Mean coherency related to alignment of collagen fibers was 0.66 ± 0.12. The crab-eating fox cornea had predominantly thick collagen fibers. Crab-eating fox eyes have morphofunctional peculiarities. They resemble the eyes of domestic dogs in some aspects, but diverge in others.

Suture and venous traction test analysis in dogs fixed in alcohol and preserved in saline solution / Análise de sutura e de teste de tração venosa em cães fixados em álcool e conservados em solução salina

The objective of this research was to determine in necropsied dogs the best time for fixation in ethylic alcohol (EA) and preservation in 30% sodium chloride aqueous solution (SCAS 30%), aiming micro-surgical training. Five groups of necropsied dogs (G1 to G5) were fixed with EA, and put in boxes containing EA for 30 (G2), 60 (G3), 90 (G4) or 120 days (G5). After that, each group was preserved in SCAS 30% for 120 days. The control group (G1) was composed by cadavers without fixation/preservation. At the end of each period, two fragments of external jugular vein per cadaver were collected, for traction test. Immediately after the collection, the cadavers femoral veins were evaluated (by 2 people) regarding the suture quality in binocular surgical microscope, and attributed scores from 0 (bad) to 5 (excellent), regarding the fresh samples. The average at the maximum rupture strength of the G3 fixation end (21.51N), such as the average of the G2 preserving end (21.62N) remained closer to the control group (19.98N) and the G2 was the group with the best score for venous suture training. The EA was efficient as a fixative just like SCAS as a dog cadavers' preservative. The small change of the traction test values, together with the best suture score, indicated the group kept for 30 days in EA and SCAS (G2) as the best for venous micro-surgical training.(AU)

O objetivo deste trabalho foi determinar, em cadáveres de cães, o melhor tempo para fixação em álcool etílico (AE) e conservação em solução aquosa de cloreto de sódio (SACS) a 30% visando o treinamento microcirúrgico. Cadáveres de cinco grupos (G1 a G5) foram fixados com AE e colocados em caixas contendo AE por 30 (G2), 60 (G3), 90 (G4) ou 120 days (G5). Depois, cada grupo foi conservado em SACS 30% por 120 dias. O grupo controle (G1) foi composto de cães sem fixação/conservação. Ao final de cada período, 2 fragmentos da veia jugular externa por cadáver foram coletados para o teste de tração. Imediatamente após a coleta, as veias femorais foram avaliadas (por 2 pessoas) em relação à qualidade da sutura em microscópio cirúrgico binocular, e atribuídos escores de 0 (péssimo) à 5 (excelente), em relação às amostras frescas. A média da força máxima de ruptura no G3 no final da fixação (21,51N), assim como a média do G2 no final da conservação (21,62N) foram as que mais se mantiveram próxima do grupo controle (19,98N) e o G2 foi o grupo com o melhor escore para treinamento da sutura venosa. O AE foi eficiente como fixador assim como a SACS foi efetiva na conservação de cadáveres de cães. A pequena alteração nos valores do teste de tração, junto com o melhor escore para sutura, indicaram o grupo mantido por 30 dias em AE e SACS (G2) como o melhor para treinamento venoso microcirúrgico.(AU)

Reduction in Histone H3 Acetylation and Chromatin Remodeling in Corneas of Alloxan-Induced Diabetic Rats.

PURPOSE: To evaluate acetylation of histone H3, chromatin remodeling, nuclear size and shape, DNA ploidy, and distribution of nucleolus organizing regions (NORs) in corneal epithelial and stromal cells of diabetic and nondiabetic rats. METHODS: Diabetes was induced by a single intraperitoneal injection of alloxan. All diabetic rats (n = 20) included in the study had 4 weeks of moderate-to-severe hyperglycemia (plasma glucose levels >400 mg/dL). Acetylated histone H3 levels were quantified in corneal tissue using a colorimetric assay. Chromatin remodeling, nuclear sizes (area/perimeter) and shapes (circularity), and DNA ploidies were evaluated from Feulgen-stained tissue sections using video image analysis. Distributions of NORs were studied in tissue sections impregnated with silver ions. Ophthalmic clinical parameters, including corneal sensitivity, were investigated. Twenty nondiabetic rats were used as controls. RESULTS: Acetylation of histone H3 was reduced in the corneas of the diabetic rats. Nuclei in corneal epithelial cells of diabetic rats compacted chromatin, increased in size, modified their shapes, and elevated DNA ploidy. The only nuclear change observed in the corneal stromal cells of diabetic rats was chromatin decompaction. The size of the silver-stained NOR did not differ between the study samples. The corneal sensitivity in diabetic rats was 51.8% lower than that in nondiabetic rats. CONCLUSIONS: The results of this study show that alloxan-induced diabetes altered the histone H3 acetylation pattern and compromised the chromatin supraorganization in corneal tissue/cells. Continued research is needed to understand the clinical and morphofunctional significance of changes in corneal cell nuclei of diabetic individuals.

Molecular characterization and potential sources of aqueous humor bacterial contamination during phacoemulsification with intraocular lens implantation in dogs.

Bacterial contamination of the anterior chamber during cataract surgery is one of the main responsible for endophthalmitis postoperative. Phacoemulsification is a less invasive technique for cataract treatment, although it does not exclude the possibility of contamination. In this study, bacterial contaminants of aqueous humor collected pre- and post-phacoemulsification with intraocular lens implantation (IOL) of twenty dogs were identified. As the conjunctival microbiota constitute a significant source of anterior chamber contamination, bacterial isolates from aqueous humor were genetically compared with those present in the conjunctival surface of the patients. Three dogs presented bacterial growth in both aqueous humor and conjunctival surface samples. Bacterial isolates from these samples were grouped according to their genetic profiles by repetitive-element PCR (rep-PCR) and their representatives were identified by 16S rRNA sequencing. Isolates from conjunctival surface were identified as Enterobacter spp., Staphylococcus spp. and S. aureus; and from aqueous humor samples as Enterobacter spp., Pantoea spp., Streptococcus spp. and Staphylococcus spp., respectively in decreasing order of prevalence. According to the rep-PCR analysis, 16.6% of Enterobacter spp. isolates from conjunctival surface were genetically similar to those from aqueous humor. The rest of isolates encountered in aqueous humor were genetically distinct from those of conjunctival surface. The significant genetic diversity of bacterial isolates found in the aqueous humor samples after surgery denoted the possibility of anterior chamber contamination during phacoemulsification by bacteria not only from conjunctival surface but also from different sources related to surgical environment.

Effects of intracameral ascorbic acid on the corneal endothelium of dogs undergoing phacoemulsification.

OBJECTIVE: Cataracts are the most common ocular disorder in dogs. Phacoemulsification is the preferred treatment method among ophthalmologists, but the cellularity of the endothelium must be considered for its success, as endothelial lesions may produce permanent corneal decompensation. The objective of this study was to evaluate the effects of intracameral ascorbic acid, a known antioxidant, on the corneal endothelium of dogs undergoing phacoemulsification. ANIMAL STUDIED: In all, 40 eyes from 20 dogs, males and females from 7 to 12 years of age, were assessed for mature cataracts. PROCEDURES: Two groups were formed (n = 20): Group 1 (G1) received a balanced salt solution (BSS), whereas Group 2 (G2) received sterile ascorbic acid diluted in a BSS, at a final concentration of 0.001 m ascorbic acid. The corneal endothelium was assessed via non-contact specular microscopy at multiple time points before and after phacoemulsification. Cell density (cells/mm2 ) and area (mm2 ), corneal thickness (mm), hexagonality, and the coefficient of variation of cell size were all assessed. P values equal to or less than 0.05 were considered significant. RESULTS: With respect to the density of endothelial cells, both groups showed losses, but they were less severe in G2. There were no differences in corneal thickness. Hexagonality decreased significantly in the postoperative period in G1. Also in G1, the coefficient of variation of cell size increased significantly. CONCLUSION: According to the results obtained, ascorbic acid minimizes cellular losses in the corneal endothelium.

Ophthalmic parameters in adult lowland paca (Cuniculus paca) raised in captivity.

OBJECTIVE: To investigate the ophthalmic parameters of lowland pacas, including the anatomic features, tear production, intraocular pressure, central corneal thickness, and morphology of the corneal endothelium. ANIMALS STUDIED: Thirteen adult, anesthetized Cuniculus paca. PROCEDURE: Eyes were evaluated using slit-lamp biomicroscopy, the Schirmer tear test I, digital applanation tonometry, binocular indirect ophthalmoscopy, and noncontact specular microscopy. RESULTS: The biomicroscopy findings showed blue/brown pigmented bulbar conjunctivae, well-developed cilia (only in the upper eyelid margin), superior and inferior lacrimal puncta, brown irides, round pupils, and vestiges of the nictitating membrane. The results of the Schirmer tear test I revealed (mean ± SD) a lacrimation rate of 4.10 ± 0.44 mm/min. The intraocular pressure was 6.34 ± 0.43 mmHg. Central corneal thickness measured by specular microscopy was 0.35 ± 0.01 mm. The mean values of density, hexagonality, and the area of the endothelial cells were 2083.15 ± 42.47 cells/mm2 , 67.07 ± 3.30%, and 486.30 ± 9.56 µm2 , respectively. CONCLUSIONS: The ocular parameters defined in this study may be used for reference in future studies and might also contribute to therapeutic approaches appropriate to this species.

Histological, morphometric, protein and gene expression analyses of rat retinas with ischaemia-reperfusion injury model treated with sildenafil citrate.

The aim of this study was to better understand the role of apoptosis in a retinal ischaemia-reperfusion injury model and to determine whether sildenafil citrate treatment can prevent retinal cell apoptosis. Thirty-six rats were divided into a control group (n = 6) and two experimentally induced ischaemia-reperfusion groups (7 and 21 days; n = 15 per group). The induced ischaemia-reperfusion groups were treated with sildenafil for 7 and 21 days (n = 10 per group), and 10 animals were treated with a placebo for the same period (n = 5 per group). Paracentesis of the anterior chamber was performed with a 30-G needle attached to a saline solution (0.9%) bag positioned at a height of 150 cm above the eye for 60 min. Intraocular pressure was measured by rebound tonometer (TonoVet® ). The eyes were analysed by histology and morphometry, and by immunohistochemistry and qRT-PCR for expression of Caspase-7, Caspase-6, Caspase-9, Tnf-r2, Fas-l, Bcl-2 and Bax. Sildenafil-treated animals showed lower levels of histopathological changes (inflammatory, cellular and tissue) than their placebo-treated counterparts at both 7 and 21 days. The retinal ganglion cell layer (RGC) was preserved in the sildenafil groups (SG), with a cell count closer to control than in the placebo groups (PG). Caspase-7 expression was significantly higher in both treated groups at 7 days compared to controls. Gene expression levels in both treatment groups differed from the controls, but in SG Bax and Caspase-6 expression levels were similar to control animals. These results suggest that the main mechanism of retinal cell death in this model is apoptosis, as there is an increase in pro-apoptotic factors and decrease in the anti-apoptotic ones. Also, sildenafil seems to protect the retinal ganglion cell layer from apoptosis. Cell survival was evident in the histological and morphometric analyses, and sildenafil treatment had a protective effect in the apoptosis pathways, with gene expression levels in SG similar to the controls.

Supra-organization and optical anisotropies of the extracellular matrix in the amniotic membrane and limbal stroma before and after explant culture.

In this research we evaluated the supramolecular organizations and the optical anisotropical properties of the de-epithelialized human amniotic membrane and rabbit limbal stroma, before and after explant culture. Birefringence, monochromatic light spectral absorption and linear dichroism of the main extracellular matrix biopolymers, that is, the fibrillar collagens and proteoglycans, were investigated by polarized light microscopy combined with image analysis. Our results demonstrated that the culture procedure-induced stimuli altered the supra-organizational characteristics (in terms of collagens/proteoglycans spatial orientation and ordered-aggregational state) of the amniotic and limbal extracellular matrix, which led to changes in optical anisotropical properties.

Effects of antiproteolytic agents on corneal epithelial viability and matrix metalloproteinase-2 and metalloproteinase-9 activity in alkali-burned corneas of rats.

OBJECTIVE: To evaluate the effects of agents on corneal re-epithelization and metalloproteinase-2 and metalloproteinase-9 (MMP-2 and MMP-9) activities in corneas of rats submitted to ulceration. ANIMALS STUDIED: Ninety eight healthy rats. PROCEDURES: Corneal ulcers were created using 1N NaOH in their left eye. Eyes were treated every 6 h with 1% ethylenediaminetetraacetic acid (EDTA), 3% chondroitin sulfate (CS), 10% N-acetylcysteine NAc and saline (S) at 6-h intervals. Corneas were stained with fluorescein and photographed at the same time points. Following 20 h and 40-42 h of corneal injury, corneas were processed for scanning electron microscopy (SEM) to quantify microvilli density, and MMPs activities were analyzed using zymography. RESULTS: The percentage of wound area and the time in hours for corneal re-epithelization did not differ significantly among treatment groups (P > 0.05). In first and the second moments, latent MMP-2 was significantly elevated in the eyes treated with NAC and CS (P < 0.001). Active MMP-2 did not change significantly among treatment groups in the first moment (P > 0.05); significantly higher activity was observed in the second moment in the eyes treated with CS (P <0.001). In the second moment, latent MMP-9 decreased significantly in eyes treated with EDTA and S (P < 0.01). Microvilli corneal density did not change significantly between healthy subjects and treatment groups (P > 0.05). CONCLUSION: Any of the studied substances did not accelerate corneal re-epithelization and did not add protection to the corneal microvilli. Significant higher levels of active form of MMP-2 in 3% chondroitin sulfate-treated group may indicate that the agent acts as substrate for such enzyme. At the end of the experiment, 1% EDTA was the most efficient agent to inhibit significantly the latent form of MMP-9. However, any of the substances add benefit over saline on reducing the proteolytic activity in the cornea of rats after alkali injury.

Light microscopic evaluation and scanning electron microscopic analysis of horse eyes following deep anterior lamellar keratectomy.

OBJECTIVE To describe the technique of deep anterior lamellar keratoplasty (DALK) with Descemet's membrane (DM) exposure in horse eyes. Also, to compare the efficacy and safety of viscodissection and big-bubble techniques for DALK. ANIMALS STUDIED Thirty-four ex vivo horse eyes. PROCEDURE Deep anterior lamellar keratoplasty was performed in 34 ex vivo horse eyes. Two groups (Group V--viscodissection--2% sodium hyaluronate; Group A--air--big-bubble) of 17 eyes were studied. Other than the substance used, the surgical technique was similar for both groups. Nonperforated eyes were submitted for light microscopic histologic evaluation and scanning electron microscopic (SEM) analysis. RESULTS Group V--Perforations occurred in 18% of the eyes during surgery. Light microscopy revealed exposure of DM in 28% of the eyes with mean thickness of the remaining stroma being 70.4 µm. Group A--Perforations occurred in 42% of the eyes. Light microscopy revealed exposure of DM in 60% of the eyes with mean thickness of the remaining stroma being 23.3 µm. No significant differences in safety, efficacy and thickness of the remaining stroma (including all eyes or excluding those with DM exposure) were observed. SEM of the surgical site revealed a more even surface in those eyes with DM exposure compared to eyes with thicker remaining stroma in both groups. CONCLUSIONs We describe two DALK techniques (viscodissection and big-bubble) for use in horses. No significant differences in safety, efficacy and thickness of the remaining stroma were observed. However, a nonsignificant trend toward the big-bubble technique being more efficacious but less safe was observed.

Expression of matrix metalloproteinases, type IV collagen, and interleukin-10 in rabbits treated with morphine after lamellar keratectomy.

PURPOSE: To study the effects of topical administration of 1% morphine on corneal analgesia in rabbits submitted to lamellar keratectomy and to assess the expression of matrix metalloproteinase-1, metalloproteinase-2, metalloproteinase-9 (MMPs), type IV collagen, and interleukin-10 (IL-10) during the treatment. METHODS: Morphine group (MG) received 50 µL of topical 1% morphine four times daily, while the control group received saline instead. Corneal touch threshold (CTT) and the wound area were assessed until corneal healing. Corneal samples were processed for routine histology, immunohistochemistry, zymography, and ELISA. RESULTS: Following keratectomy, CTT increased significantly from 6 to 96 h time points. Mean corneal re-epithelization rate and scores of leukocyte infiltration did not differ significantly between treatment groups. Immunolabeling pattern for MMP-1, MMP-9, and type IV collagen was similar in both treatment groups. In the MG, zymography indicated significantly higher levels of active MMP-2 on days 6 and 12; and in the latent MMP-9, on days 3 and 6, and in the active MMP-9, on day 6. Latent MMP-2 and MMP-9, and active MMP-9 decreased to values close to those of healthy corneas on day 12, but levels of active MMP-2 remained significantly elevated in the MG. IL-10 levels measured on days 1-6 were reduced as compared to those of healthy corneal tissue and returned to levels close to those of healthy corneas on day 12. CONCLUSION: Topical morphine promoted corneal analgesia for up to 4 days and did not delay corneal re-epithelization. The re-establishment of MMPs and IL-10 to levels close to baseline values at the end of the study and the expression of type IV collagen in both groups reinforce that, with caution, 1% morphine can be used after lamellar keratectomy in rabbits.

Computed tomographic-dacryocystography (CT-DCG) of the normal canine nasolacrimal drainage system with three-dimensional reconstruction.

The aims of the study were (1) to quantify the influence of selected imaging parameters on the image quality (slice thickness, mAs, and beam orientation) defining optimal conditions for scan protocols and (2) to evaluate the benefits of the 3D reconstruction techniques for visualization of NDS structures in dogs. CT-DCG was performed bilaterally in 32 heads of dogs. CT transverse images were obtained using a combination of two slice thickness values (0.8 mm and 2 mm) and two mAs values (50 mAs and 300 mAs). Two beam projection orientations were also tested: transverse plane (perpendicular to the hard palate) and oblique to the hard palate. Three-dimensional images were obtained using Volume Rendering (VR). Transverse beam projection proved to be superior for the assessment of the inferior and superior lacrimal canaliculi and lacrimal sac. In this study, there was no statistical difference regarding mAs values (50 mAs and 300 mAs) and slice thickness values (0.8 mm and 2 mm). Three-dimensional images were helpful for the assessment of topographic relationship between nasolacrimal structures and cranial landmarks.

Use of specular microscopy to determine corneal endothelial cell morphology and morphometry in enucleated cat eyes.

The purpose of this study was to investigate the effect of age on endothelial morphology and morphometry in cats. The corneal endothelium was studied using a contact specular microscope. A total of 18 cats (Felis catus Linnaeus, 1758) were evaluated in this study. The subjects were divided into three groups of six cats each in function of age: G1 (1 to 3 months old), G2 (5 to 12 months old), and G3 (24 to 40 months old). The examination presented data as endothelial cell density (ECD), average cell area, corneal thickness, polymegathism, and pleomorphism. Results revealed ECD decrease in corneas of normal cats with age, as well as a corresponding increase in endothelial cell area and pleomorphism. The present work suggests that the endothelial parameters evaluated change with advancing age.

Ultrassonografia em modo A e B na avaliação pré-operatória da lente e do segmento posterior de olhos de cães com catarata / A and B mode ultrasonography in preoperative evaluation of lens and posterior segment of dogs eyes with cataract

Ultrasonography of the lens and posterior segment is an indispensable step in the preoperative evaluation of dogs with cataracts, since ophthalmoscopy is not feasible when there is opacification of the lens. This study evaluated the echographic conditions of cataractous lens and fundus of the eye in dogs affected by cataracts. The study was conducted in 30 dogs (56 eyes), 10 males and 20 females, with different types of cataracts at different stages of development. Echography in A and B modes, simultaneously, was carried out for the examination of the lens and posterior segment. The examinations revealed anterior cortical, posterior cortical and nuclear cataract in 12 eyes (21.4 percent), anterior cortical, posterior cortical, nuclear and posterior capsular in 23 eyes (41 percent), anterior cortical, posterior cortical and posterior capsular cataract in one eye (1.7 percent), anterior cortical and nuclear cataract in one eye (1.7 percent), anterior cortical, nuclear and posterior capsular cataract in five eyes (8.9 percent), and anterior cortical cataract in seven eyes (12.5 percent). Abnormal ultrasonographic alterations were observed in the posterior segment in 26 eyes evaluated (46.4 percent). Vitreal degeneration was detected in 12 eyes (21.4 percent), images of vitreal exudate or hemorrhage in seven eyes (12.5 percent), persistence of hyaloid artery in four eyes (7.1 percent) and lens subluxation in three eyes (5.3 percent). The results obtained reiterate the importance of ultrasonography in canine patients presented for cataract surgery given that alterations of the posterior segment are difficult to identify in a clinical examination when the lens is opacified.

A ultrassonografia do segmento posterior do bulbo do olho é etapa indispensável na avaliação de cães com catarata que serão submetidos à facectomia, uma vez que a oftalmoscopia não é factível quando há opacificação da lente, notadamente nas cataratas maduras. Este estudo avaliou as condições ecográficas da lente cataratogênica e do fundo de olho de cães cursando com catarata. Foram avaliados 30 cães (56 olhos), sendo 10 machos e 20 fêmeas, cursando com catarata em diferentes estágios de desenvolvimento. Realizou-se a ecografia em modo A e B simultaneamente. Os exames revelaram catarata cortical anterior, posterior e nuclear em 12 olhos (21,4 por cento), catarata cortical anterior, posterior, nuclear e capsular posterior em 23 olhos (41 por cento), catarata cortical anterior, posterior e capsular posterior em um olho (1,7 por cento), catarata cortical anterior e nuclear em um olho (1,7 por cento), catarata cortical anterior, nuclear e capsular posterior em cinco olhos (8,9 por cento) e catarata cortical anterior em sete olhos (12,5 por cento). Alterações no segmento posterior foram observadas, à ultrassonografia, em 26 olhos avaliados (46,4 por cento). Degeneração vítrea foi detectada em 12 olhos (21,4 por cento), imagens compatíveis com exsudato ou hemorragia em sete olhos (12,5 por cento), persistência da artéria hialóide em quatro olhos (7,1 por cento) e subluxação da lente em três olhos (5,3 por cento). Os resultados obtidos reiteram a importância da ultrassonografia em cães candidatos à cirurgia de catarata, uma vez que o exame do segmento posterior do bulbo do olho é de difícil execução, devido à opacificação da lente.

Ultrasonographic and echobiometric findings in the eyes of Saanen goats of different ages.

OBJECTIVE: To determine ultrasonographic and echobiometric findings in the eyes of goats of different ages. B and A mode ultrasonography was performed in 55 healthy Saanen goats (110 eyes) of ages 45 (n = 15), 180 (n = 20), and 549 days (n = 20). PROCEDURES: Transcorneal ultrasonography was performed after instillation of a topical anesthetic and application of acoustic transmission gel on the 20-MHz linear transducer tip. The transducer was placed in a longitudinal position until optimal B scan images, according to echoes of A mode images, were obtained. Statistical analysis was carried out to compare the echobiometric readings among subjects (P < 0.05). RESULTS: The aqueous and vitreous humors of the goats, as well as the lens cortices and nuclei, appeared anechoic. The cornea, anterior and posterior lens capsule, sclera, and iris appeared hyperechoic and generated peaks on the A mode. Ocular measurements between the right and left eyes, as well as between male and female goats were not significantly different among subjects of the same age (P = 1.00). All dimensions evaluated increased significantly with age (P < 0.01). CONCLUSIONS: The ultrasonographic appearances of goat eyes were very similar to those of other animal species. The cornea, anterior chamber, lens, vitreous chamber, and axial globe length increased with age in the Saanen breed of goats. Knowledge of the normal appearance and ocular dimensions of goat eyes facilitates the use of ultrasonography in the evaluation of ocular disease in Saanen goats.

Conjunctival effects of canine distemper virus-induced keratoconjunctivitis sicca.

OBJECTIVE: This study compared the histopathology of canine distemper virus (CDV)-induced keratoconjunctivitis sicca (KCS) to non-infectious KCS in conjunctival tissues. ANIMALS STUDIED: Forty mongrel dogs were assigned to three distinct groups: (i) non-infectious KCS (G1, n = 10), (ii) CDV-induced KCS (G2, n = 20), and (iii) healthy animals without any ocular alterations (G3, n = 10). PROCEDURE: IgG titers and physical and ophthalmic examinations (e.g. Schirmer tear test [STT], tonometry, biomicroscopy, indirect biomicroscopy, and fluorescein test) were performed on all dogs. Conjunctival biopsies were collected and examined microscopically. RESULTS: Non-infectious and CDV-induced KCS demonstrated similar histopathological changes. Both types of KCS correlated with low STT, conjunctival hyperemia, mucopurulent ocular discharge, predominant lymphoplasmacytic infiltration, and acantholysis and keratinization of the ocular surface. G1 had lower conjunctival goblet cell counts than G3. Inclusion bodies were sporadically found in conjunctival samples of dogs from G2. The severity of ocular lesions in G1 and G2 did not correlate with the histopathological findings. CONCLUSIONS: Dogs with non-infectious and CDV-induced KCS had very similar conjunctival histopathology. Our findings suggest that the pathophysiology of CDV-induced KCS is likely to be the same as non-infectious KCS, that is, a result of lacrimal deficiency and inflammation of the ocular surface.

Morphological analysis of the corneal endothelium in eyes of dogs using specular microscopy / Análise morfológica com microscopia especular do endotélio corneal em olhos de cães

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.

Foram estudados 20 olhos de 10 cães sem raça definida, machos e fêmeas com 6 anos de idade. A morfologia das células do endotélio da córnea foi analisada utilizando-se microscópio especular de contato. Foram estudadas as regiões central e periférica da córnea. De cada região da córnea foram realizadas três micrografias. De cada micrografia foram analisadas no mínimo 100 células endoteliais. Foram obtidos os valores do polimegatismo e pleomorfismo. O endotélio corneano de cães caracterizou-se por uma monocamada de células poligonais uniformes em tamanho e forma. A forma predominante das células endoteliais foi hexagonal. O índice de polimegatismo foi 0.22. Este estudo demonstrou que a morfologia das células do endotélio da córnea de cães é semelhante à observada em humanos.