Comparison of Electrolyte Composition and Crystallization Patterns in Bird and Reptile Tears.

To compare tear electrolytes and tear crystallization patterns in birds and reptiles, tears were sampled by Schirmer tear test from 10 animals each of Ara ararauna, Amazona aestiva, Tyto alba, Rupornis magnirostris, Chelonoidis carbonaria, and Caiman latirostris, and 5 of Caretta caretta. The aliquots were pooled to assess concentrations of total protein, chloride, phosphorus, iron, sodium, potassium, calcium, and urea. For the tear ferning test, samples of each species were observed under a polarized light microscope at room temperature and humidity. Crystallization patterns were graded according Rolando and Masmali scales. There was more total protein and urea in owl and sea turtle tears, respectively, than in the other animals tested. Electrolyte balance was similar for all species, with higher sodium, chloride, and iron. In birds, Rolando-scale grades of tear crystallization patterns ranged from I to II, and from 0 to 2 using the Masmali scale; in reptiles, grades were II to IV (Rolando) and 2 to 4 (Masmali). Crystallization arrangements of some species had higher scores, as caimans and sea turtles, possibly due to different the tear composition. Marine and lacustrine species presented higher. The ionic balance of lacrimal fluids of birds and reptiles was similar to that in humans, with higher values of sodium and chloride. However, a similar tear composition did not influence the crystal morphology. Crystallization classification suggested that higher grades and types are due to the different microelements present in the tears of wild species.

Evaluation of lacrimal production, osmolarity, crystallization, proteomic profile, and biochemistry of capuchin monkeys' tear film.

BACKGROUND: To evaluate the composition and characteristics of capuchin monkey (CM) tear film. METHODS: Eleven CM (Sapajus sp.) was evaluated. Strip meniscometry test (SMT), osmolarity, and tear ferning test (TFT) (by Rolando and Masmali scales) were assessed. Tear protein profile (SDS-PAGE), and total protein, albumin, urea, glucose, and cholesterol levels in tear film and blood serum were screened. RESULTS: Median ± semi-interquatil range for SMT and osmolarity values were 8.0 ± 1.625 and 303.0 ± 9.875, respectively. TFT for Rolando was 2.0 ± 0.5, and Masmali was 2.0 ± 0.0. Monkeys's tear obtained type II and III for Rolando, and 1 and 2 for Masmali. Tear components showed protein bands among 23-217 kDa, and presence of albumin, urea, glucose, and cholesterol. CONCLUSIONS: The results of SMT, osmolarity, TFT, SDS-PAGE, and tear biochemistry may serve as a reference baseline for CM, and the data may serve as a basis for future experimental model evaluations.

Utilidad de la proteína de unión al factor de crecimiento similar a la insulina tipo 1 en la ruptura prematura de membrana / Utility of insulin-like growth factor-binding protein type 1 in premature membrane rupture

Introducción: Fercuentemente no disponemos de una clara evidencia de la pérdida de líquido amniótico observado por examen con espéculo, por lo que el diagnóstico de rotura prematura de membranas puede ser con frecuencia incierto, por lo que se necesitan pruebas de diagnóstico apropiadas y complementarias para la toma de decisiones. Objetivo: conocer la precisión diagnóstica de la proteína-1 de unión al factor de crecimiento similar a la insulina (IGFBP-1) en la rotura prematura de membranas al compararla con la medición del bolsilla mayor por ecografía y el Test de de Ferning. Material y métodos: 102 gestantes de 24 a 37 semanas con signos y/o síntomas de rotura de membranas fueron elegibles, fueron evaluadas con las pruebas IGFBP-1, ecografía y Test de Ferning. Resultados: Para el IGFBP-1 se obtuvo 95% de sensibilidad (S), 95% de especificidad (E), 95% de valor predictivo positivo (VPP) y 96% de valor predictivo negativo (VPN). Par el Test de Fernig se obtuvieron valores de 85%, 25%, 25% y 83% respectivamente. En tanto que para la ecografía los hallazgos fueron de 81%, 29%, 56% y 58% respectivamente. Conclusión: el ensayo IGFBP-1 fue el método más preciso para diagnosticar la ruptura prematura de membranas con la mayor sensibilidad, especificidad, valor predictivo positivo y valor predictivo negativo.

Introduction: We do not have clear evidence of the loss of amniotic fluid observed by speculum examination, so the diagnosis of premature rupture of membranes can often be uncertain and appropriate and complementary diagnostic tests are needed for decision making. Objective: to know the diagnostic accuracy of insulin-like growth factor-binding protein-1 (IGFBP-1) in the premature rupture of membranes when compared with the measurement of the greater pocket by ultrasound and the Ferning test. Material and methods: 102 pregnant women from 24 to 37 weeks with signs and / or symptoms of rupture of membranes were eligible, which were evaluated with the IGFBP-1, ultrasound and Ferning Test. Results: For IGFBP-1 95% sensitivity (S), 95% specificity (E), 95% positive predictive value (PPV) and 96% negative predictive value (NPV) were obtained. For the Ferning Test values of 85%, 25%, 25% and 83% respectively were obtained. While for ultrasound the findings were 81%, 29%, 56% and 58% respectively. Conclusion: the IGFBP-1 assay was the most accurate method to diagnose the premature rupture of membranes with the highest sensitivity, specificity, positive predictive value and negative predictive value.

Progress in tear microdesiccate analysis by combining various transmitted-light microscope techniques.

BACKGROUND: Tear desiccation on a glass surface followed by transmitted-light microscopy has served as diagnostic test for dry eye. Four distinctive morphological domains (zones I, II, III and transition band) have been recently recognized in tear microdesiccates. Physicochemical dissimilarities among those domains hamper comprehensive microscopic examination of tear microdesiccates. Optimal observation conditions of entire tear microdesiccates are now investigated. One-µl aliquots of tear collected from individual healthy eyes were dried at ambient conditions on microscope slides. Tear microdesiccates were examined by combining low-magnification objective lenses with transmitted-light microscopy (brightfield, phase contrasts Ph1,2,3 and darkfield). RESULTS: Fern-like structures (zones II and III) were visible with all illumination methods excepting brightfield. Zone I was the microdesiccate domain displaying the most noticeable illumination-dependent variations, namely transparent band delimited by an outer rim (Ph1, Ph2), homogeneous compactly built structure (brightfield) or invisible domain (darkfield, Ph3). Intermediate positions of the condenser (BF/Ph1, Ph1/Ph2) showed a structured roughly cylindrical zone I. The transition band also varied from invisibility (brightfield) to a well-defined domain comprising interwoven filamentous elements (phase contrasts, darkfield). CONCLUSIONS: Imaging of entire tear microdesiccates by transmitted-light microscopy depends upon illumination. A more comprehensive description of tear microdesiccates can be achieved by combining illumination methods.

Tear ferning test in horses and its correlation with ocular surface evaluation.

OBJECTIVE: To describe the tear ferning test (TFT) in healthy horses and its correlation with other parameters for evaluating the ocular surface. ANIMALS STUDIED: Thirty male and female adult healthy horses (60 eyes), of no defined breed. PROCEDURES: Tear sample was collected with a microcapillary tube, placed on the surface of a glass slide, and allowed to dry at room temperature. The crystallization pattern was classified according to Rolando (Chibret International Journal Ophthamology, 1984; 2, 32). The program STEPanizer(©) stereology tool, version 1.0, was utilized for counting points on the digitally captured crystallization image. A conjunctival biopsy was performed. RESULTS: Tear ferning test was classified as Type I in 18 eyes (30%), Type II in 31 eyes (51.7%), and Type III in 11 eyes (18.3%), at a mean temperature of 27.3 ± 1.5 °C and relative humidity of 61.5 ± 5.7%. In the Type I crystallization, the count varied between 27 and 36 points (mean: 33.27 ± 2.40), in Type II between 22 and 31 points (25.42 ± 1.95), and in Type III between 13 and 25 points (16.82 ± 3.76). There was no statistical difference or correlation between the right and left eyes, nor was there a statistically significant influence (P < 0.05) on TFT by the factors evaluated. The mean goblet cells values were 50 ± 11.4 cells/field. All samples showed the presence of lymphocytes, plasmocytes, and eosinophils. CONCLUSION: Tear ferning test is easy to perform, without risks to the patient. Once standardized for horses, associated or not with the program STEPanizer(©) stereology tool, it is an additional method for evaluating the ocular surface.

Progress in tear microdesiccate analysis by combining various transmitted-light microscope techniques

BACKGROUND: Tear desiccation on a glass surface followed by transmitted-light microscopy has served as diagnostic test for dry eye. Four distinctive morphological domains (zones I, II, III and transition band) have been recently recognized in tear microdesiccates. Physicochemical dissimilarities among those domains hamper comprehensive microscopic examination of tear microdesiccates. Optimal observation conditions of entire tear microdesiccates are now investigated. One-µl aliquots of tear collected from individual healthy eyes were dried at ambient conditions on microscope slides. Tear microdesiccates were examined by combining low-magnification objective lenses with transmitted-light microscopy (brightfield, phase contrasts Ph1,2,3 and darkfield. RESULTS: Fern-like structures (zones II and III) were visible with all illumination methods excepting brightfield. Zone I was the microdesiccate domain displaying the most noticeable illumination-dependent variations, namely transparent band delimited by an outer rim (Ph1, Ph2), homogeneous compactly built structure (brightfield) or invisible domain (darkfield, Ph3). Intermediate positions of the condenser (BF/Ph1, Ph1/Ph2) showed a structured roughly cylindrical zone I. The transition band also varied from invisibility (brightfield) to a well-defined domain comprising interwoven filamentous elements (phase contrasts, darkfield. CONCLUSIONS: Imaging of entire tear microdesiccates by transmitted-light microscopy depends upon illumination. A more comprehensive description of tear microdesiccates can be achieved by combining illumination methods.

Dynamics of tear fluid desiccation on a glass surface: a contribution to tear quality assessment

BACKGROUND: Fern-like crystalloids form when a microvolume of tear is allowed to dry out at ambient conditions on a glass surface. Presence of crystalloids in tear "microdesiccates" is used to evaluate patients with Dry-Eye disease. This study aims to examine morphologically the desiccation process of normal tear fluid and to identify changes associated with accelerated tear evaporation. Tear microdesiccates from healthy (Non-Dry Eye) and Dry Eye subjects were produced at ambient conditions. Microdesiccate formation was monitored continuously by dark-field video microscopy. Additionally, accelerated desiccation of tear samples from healthy subjects was conducted under controlled experimental conditions. Particular morphological domains of tear microdesiccates and their progressive appearance during desiccation were compared. RESULTS: In normal tear microdesiccates, four distinctive morphological domains (zones I, II, III and transition band) were recognized. Stepwise formation of those domains is now described. Experimentally accelerated desiccation resulted in marked changes in some of those zones, particularly involving either disappearance or size reduction of fern-like crystalloids of zones II and III. Tear microdesiccates from Dry Eye subjects may also display those differences and be the expression of a more synchronous formation of microdesiccate domains. CONCLUSION: Morphological characteristics of tear microdesiccates can provide insights into the relative rate of tear evaporation.

Microdesiccates produced from normal human tears display four distinctive morphological components

Desiccation of human tears on glass surfaces results in fern-like crystalloids. This phenomenon has been associated with tear normality (Tear Ferning Test, TFT) and is used as a diagnostic aid to evaluate patients with Dry-Eye disease. However, TFT is focused on the assessment of only a minor fraction of desiccated tear samples and considers only the relative abundance and density of fern-like crystalloids. The aim of this study was to characterize morphologically entire desiccated micro volumes of tears from healthy donors. Tear samples were collected from 23 healthy young adult volunteers. Tear aliquots (1-3 μL) were allowed to dry on glass surfaces under ambient conditions of temperature (15-25°C) and relative humidity (40-45%). Dry samples were analyzed by dark-field microscopy. Morphometric data were acquired with Image J software. Tear volume was positively correlated with both area and time of desiccation. Morphological features of multiple microdesiccates produced from a single subject displayed striking similarities whereas tear microdesiccates from different healthy subjects displayed consistent differences but shared a common general design. This design may be mostly represented by the occurrence of four distinctive zones, named as zones I, II, III and Transition band. The main features of these zones are described.