Impact of human coronavirus infections on paediatric patients at a tertiary paediatric hospital: a retrospective study of the prepandemic era.

BACKGROUND: Human coronaviruses (HCoVs) are important respiratory pathogens in humans and animals. Most HCoVs are emerging pathogens, with five known human pathogens identified in the last two decades. AIM: To examine the clinical course of HCoV infection in children to improve understanding of severity and outcomes. METHODS: A retrospective review was undertaken of all encounters of children with known HCoV infection at a tertiary paediatric hospital from January 2015 to January 2018. Electronic medical records were reviewed for demographic data, HCoV type, viral co-pathogens, time to testing, need for hospitalization, requirement for higher-level care (HLC) including intensive care unit management and requirement for oxygen support, radiographic findings suggestive of lower respiratory tract (LRT) disease, and length of stay (LOS). FINDINGS: In total, 450 encounters for 430 different patients were identified, with the majority (85%) being inpatient. OC43 was the most common HCoV. Younger patients (age <5 years) had higher probability of hospitalization [adjusted odds ratio (aOR) 2.2, 95% confidence interval (CI) 1.2-4.1], requirement for HLC (aOR 1.8, 95% CI 1.0-3.1) and presence of LRT findings on chest radiographs (aOR 1.7, 95% CI 1.01-2.9). Clinical outcomes did not differ between HCoV types, except LOS which was longer for 229E. Fifty-two (11%) encounters were detected after 3 days of hospitalization (median 25.5 days), suggesting possible nosocomial infection. CONCLUSION: HCoVs are important respiratory pathogens in the paediatric population, especially among patients aged <5 years who are at increased risk for severe disease. The role of HCoVs as hospital-acquired pathogens may be underappreciated.

In vitro antifungal susceptibility of Fusarium species and Aspergillus fumigatus cultured from eleven horses with fungal keratitis.

PURPOSE: To examine the relationship between Minimum Inhibitory Concentration (MICs) and response to therapy of 6 Fusarium spp. and 5 Aspergillus fumigatus isolated from equine ulcerative keratitis cases. PROCEDURE: Fungi were identified by morphology and Internal Transcribed Spacer (ITS) polymerase chain reaction (PCR) with sequencing and evaluated at the University of Texas Fungal Testing Laboratory for susceptibility to three azole antifungals (miconazole, voriconazole, posaconazole), natamycin, and two echinocandins (anidulafungin, caspofungin). A Mann-Whitney rank sum test was used for the comparison of time to heal between infections of different fungal genera and in vitro susceptibility to the drug administered. RESULTS: Fusarium spp. were resistant to azole antifungals in 6/6 cases (100%). Fusarium spp. were susceptible to echinocandins and natamycin in all cases. A. fumigatus was resistant to anidulafungin in 1/5 cases (20%) and posaconazole in 1/5 cases (20%) The remainder of A. fumigatus isolates were susceptible to all antifungal agents tested. Fusarium isolates were treated with antifungals to which they were not susceptible; however, all cases of A. fumigatus were treated with antifungals to which they were susceptible. All Fusarium cases and A. fumigatus cases experienced clinical resolution, regardless of surgical intervention. There was no statistical correlation between fungal genus and time to heal (p < .082). CONCLUSIONS: The in vitro susceptibility indicated that all cases of Fusarium spp. were resistant to azole antifungal drugs which were used as treatment. Clinical outcomes, however, showed that all cases healed despite resistance to antifungals.

Genetic divergence among threespine stickleback that differ in nuptial coloration.

Sexual signals are shaped by their intended and unintended receivers as well as the signalling environment. This interplay between sexual and natural selection can lead to divergence in signals in heterogeneous environments. Yet, the extent to which gene flow is restricted when signalling phenotypes vary across environments and over what spatial scales remains an outstanding question. In this study, we quantify gene flow between two colour morphs, red and black, of freshwater threespine stickleback fish (Gasterosteus aculeatus). We capitalize on the very recent divergence of signalling phenotypes in this system to characterize within-species and among-morph genetic variation and to test for levels of gene flow between colour morphs in Oregon and Washington. Despite limited evidence for assortative mating between allopatric red and black populations, we found that black populations are genetically distinct from nearby red populations and that the black morph appears to have evolved independently at least twice in Oregon and Washington. Surprisingly, we uncovered a group of stickleback in one small coastal stream, Connor Creek, which is genetically and morphologically distinct from the red and black colour morphs and from marine stickleback. Historically, both colour morphs have coexisted in this location and sometimes hybridized, raising new questions about the origins and history of these fish, which were first described as anadromous-black hybrids >50 years ago. Understanding how genetic variation is currently partitioned within and among populations and colour morphs in this system should prompt future studies to assess the relative roles of habitat, ecological and pre- and post-reproductive barriers in the genetic divergence and phenotypic patterns we observe in nature.

Therapeutic effects of equine amniotic membrane suspension on corneal re-epithelialization and haze in a modified lagomorph ex vivo wound healing model.

OBJECTIVE: To investigate the therapeutic effects of topical equine amniotic membrane (eAM) suspension following corneal wounding in a controlled experimental setting. PROCEDURES: Equine amniotic membrane was collected, gamma irradiated, homogenized for topical suspension preparation, and cryopreserved. Corneoscleral rims harvested from fresh rabbit globes were wounded via keratectomy and were maintained in an air-liquid interface ex vivo corneal culture model. Treatment groups included topical gamma irradiated eAM suspension (n = 20) and a control group (n = 20). Re-epithelialization of the wound was assessed with daily photographic evaluation of area of fluorescein uptake (mm2 ). Corneal wound haze after a 21-day period was assessed by photographic analysis of haze area (mm2 ) and pixel intensity (0-255). Histologic processing of corneal tissue was performed, and protein identification of eAM suspension using Liquid chromatography-mass spectrometry (LC-MS). RESULTS: The average day of complete corneal re-epithelialization in controls (5.5 ± 1.1) and topically treated (5.5 ± 0.6) corneas, and rates of reduction in area of fluorescein uptake over time did not significantly differ (p = .44). The corneal wound haze was significantly reduced in mean area by approximately 52% and intensity by 57% in corneas treated with topical eAM suspension (p < .05), compared to controls 21 days following wounding. Protein analysis identified numerous proteins, specifically decorin, dermatopontin, and lumican, which have previously been documented in eAM. CONCLUSIONS: Area and intensity of corneal wound haze were significantly reduced in corneas treated with gamma irradiated eAM suspension, which may be due to previously identified therapeutic proteins which promote corneal clarity.

Development of a Multi-Study Repository to Support Research on Veteran Health: The VA Cooperative Studies Program Epidemiology Center-Durham (CSPEC-Durham) Data and Specimen Repository.

Federal agencies, including the Department of Veterans Affairs (VA), have prioritized improved access to scientific data and results collected through federally funded research. Our VA Cooperative Studies Program Epidemiology Center in Durham, North Carolina (CSPEC-Durham) assembled a repository of data and specimens collected through multiple studies on Veteran health issues to facilitate future research in these areas. We developed a single protocol, request process that includes scientific and ethical review of all applications, and a database architecture using metadata (common variable descriptors) to securely store and share data across diverse studies. In addition, we created a mechanism to allow data and specimens collected through older studies in which re-use was not addressed in the study protocol or consent forms to be shared if the future research is within the scope of the original consent. Our CSPEC-Durham Data and Specimen Repository currently includes research data, genomic data, and study specimens (e.g., DNA, blood) for three content areas: colorectal cancer, amyotrophic lateral sclerosis, and Gulf War research. The linking of the study specimens and research data can support additional genetic analyses and related research to improve Veterans' health.

Phenotypic divergence among threespine stickleback that differ in nuptial coloration.

By studying systems in their earliest stages of differentiation, we can learn about the evolutionary forces acting within and among populations and how those forces could contribute to reproductive isolation. Such an understanding would help us to better discern and predict how selection leads to the maintenance of multiple morphs within a species, rather than speciation. The postglacial adaptive radiation of the threespine stickleback (Gasterosteus aculeatus) is one of the best-studied cases of evolutionary diversification and rapid, repeated speciation. Following deglaciation, marine stickleback have continually invaded freshwater habitats across the northern hemisphere and established resident populations that diverged innumerable times from their oceanic ancestors. Independent freshwater colonization events have yielded broadly parallel patterns of morphological differences in freshwater and marine stickleback. However, there is also much phenotypic diversity within and among freshwater populations. We studied a lesser-known freshwater "species pair" found in southwest Washington, where male stickleback in numerous locations have lost the ancestral red sexual signal and instead develop black nuptial coloration. We measured phenotypic variation in a suite of traits across sites where red and black stickleback do not overlap in distribution and at one site where they historically co-occurred. We found substantial phenotypic divergence between red and black morphs in noncolor traits including shape and lateral plating, and additionally find evidence that supports the hypothesis of sensory drive as the mechanism responsible for the evolutionary switch in color from red to black. A newly described third "mixed" morph in Connor Creek, Washington, differs in head shape and size from the red and black morphs, and we suggest that their characteristics are most consistent with hybridization between anadromous and freshwater stickleback. These results lay the foundation for future investigation of the underlying genetic basis of this phenotypic divergence as well as the evolutionary processes that may drive, maintain, or limit divergence among morphs.

Predator-induced maternal and paternal effects independently alter sexual selection.

Parental experience alters survival-related phenotypes of offspring in both adaptive and nonadaptive ways, yielding rapid inter- and transgenerational fitness effects. Yet, fitness comprises survival and reproduction, and parental effects on mating decisions could alter the strength and direction of sexual selection, affecting long-term evolutionary trajectories. We used a full factorial design in which threespine stickleback (Gasterosteus aculeatus) mothers, fathers, both, or neither were exposed to a model predator at developmentally appropriate times to test for predator-induced maternal, paternal, and joint parental effects on daughters' mating behavior. We tested the responsiveness, preferences, and mate choices of adult daughters in no-choice trials with wild-caught males who had varied sexual signals. Maternal and paternal predator exposure independently yielded daughters who preferred males who were intermediate in conspicuousness (with duller nuptial coloration and who courted less vigorously), relaxing the typical preference for the most conspicuous males. The combined effects of maternal and paternal predator exposure were not cumulative; when both parents were predator exposed, single-parent effects on mate preferences were reversed. Thus, we cannot assume that maternal and paternal effects additively combine to produce "parental" effects. Further, joint parental predator exposure yielded daughters who were three times less likely to mate at all. Stress-induced intergenerational parental effects on reproductive decisions such as those observed here may potentiate rapid transgenerational responses to novel and changing mating environments.

Equine ulcerative keratitis with furrow: A review of the outcomes of 72 cases from 1987 to 2015.

BACKGROUND: Ulcerative keratitis with peripheral furrow formation is a poorly-described condition which has been associated with a grave prognosis due to rapid necrosis of the cornea. OBJECTIVE: To describe the infectious aetiologies associated with furrow-forming ulcerative keratitis, its overall clinical course and the efficacy of medical and surgical intervention in horses. STUDY DESIGN: Retrospective clinical case series. METHODS: Medical records of 72 horses which presented with furrow-forming ulcerative keratitis at the University of Florida between 1987 and 2015 were reviewed. RESULTS: Seventy-two horses (72 eyes) with furrow-forming ulcerative keratitis were treated at the University of Florida between 1987 and 2015. Of these, a definitive aetiologic diagnosis was available for 37 eyes. Ten of 37 eyes (27%) were diagnosed with fungal keratitis based on cytology of corneal scraping, culture, histopathology and/or fungal PCR. Fourteen of 37 eyes (38%) were diagnosed with a mixed fungal and bacterial keratitis. Thirteen of 37 eyes (35%) were diagnosed with bacterial keratitis. Overall, 26 of 72 total eyes were treated with medical therapy alone (36%). Forty-six of 72 eyes were treated medically and surgically (64%). Of the 26 eyes which received medical therapy, 20 healed with a positive visual outcome (77%) and 6 eyes were non-visual (23%). Of the 46 eyes which received surgical intervention, 40 healed with a positive visual outcome (87%), while six eyes were non-visual (13%). Altogether, 60 of 72 eyes healed with a positive visual outcome following medical or surgical treatment of furrow-forming ulcerative keratitis (83%). Twelve of 72 eyes failed treatment (17%), with six eyes requiring enucleation and six globes becoming phthisical after treatment. MAIN LIMITATIONS: Inconsistencies in available medical record data due to the large span of time (1987-2015) are inherent in this retrospective study, along with gradual evolution of corneal surgical techniques and medical therapies over the decades. CONCLUSIONS: Furrow-forming ulcerative keratitis was associated with a positive visual outcome in 83% of horses treated at the University of Florida between the years 1987 and 2015. Furrow formation may be associated with either fungal or bacterial infection.

Color and behavior differently predict competitive outcomes for divergent stickleback color morphs.

Our knowledge of how male competition contributes to speciation is dominated by investigations of competition between within-species morphs or closely related species that differ in conspicuous traits expressed during the breeding season (e.g. color, song). In such studies, it is important to consider the manner in which putatively sexually selected traits influence the outcome of competitive interactions within and between types because these traits can communicate information about competitor quality and may not be utilized by homotypic and heterotypic receivers in the same way. We studied the roles of breeding color and aggressive behaviors in competition within and between two divergent threespine stickleback Gasterosteus aculeatus color types. Our previous work in this system showed that the switch from red to black breeding coloration is associated with changes in male competition biases. Here, we find that red and black males also use different currencies in competition. Winners of both color types performed more aggressive behaviors than losers, regardless of whether the competitor was of the same or opposite color type. But breeding color differently predicted competitive outcomes for red and black males. Males who were redder at the start of competition were more likely to win when paired with homotypic competitors and less likely to win when paired with heterotypic competitors. In contrast, black color, though expressed in the breeding season and condition dependent, was unrelated to competitive outcomes. Placing questions about the role of male competition in speciation in a sexual signal evolution framework may provide insight into the "why and how" of aggression biases and asymmetries in competitive ability between closely related morphs and species.

Assessment of Topical Therapies for Improving the Optical Clarity Following Stromal Wounding in a Novel Ex Vivo Canine Cornea Model.

Purpose: To evaluate the effect of topical suberanilohydroxamic acid (SAHA) and 5-methyl-1-phenyl-2[1H]-pyridone (pirfenidone) on the degree of corneal haze in the stromal wounded ex vivo canine cornea. Methods: Twenty-four corneoscleral rims from normal dogs were uniformly wounded with an excimer laser and placed into culture medium with an air-liquid interface. The control group (n = 8) contained placebo-treated corneas. Treatment group 1 (n = 8) received SAHA topically every 6 hours. Treatment group 2 (n = 8) received pirfenidone topically every 6 hours. Each cornea was fluorescein stained and macrophotographed every 6 hours to assess epithelialization rate. All corneas were also macrophotographed weekly to assess optical clarity (haze). Images were analyzed for differences in pixel intensity between wounded (haze) and unwounded (nonhaze) regions, and haze surface area for each cornea was calculated. Results: The mean epithelialization time was 47.25 hours in the control group, 45.00 hours in the SAHA group, and 43.50 hours in the pirfenidone group, revealing no significant difference (P = 0.368). The median difference in pixel intensity between haze and nonhaze areas was 21.5 in the control group, 8.0 in the SAHA group, and 8.0 in the pirfenidone group, which is significant (P < 0.01). The median haze surface area was 12.96 mm2 in the control group, 5.70 mm2 in the SAHA group, and 5.92 mm2 in the pirfenidone group, which is significant (P < 0.01). Conclusions: Stromal-wounded ex vivo canine corneas exhibited greater optical clarity when treated with SAHA and pirfenidone than when placebo treated at 21 days. There was no significant difference in epithelialization rate between groups. Corneal contour was correlated with geographic haze distribution.

Evaluation of three hand-held tonometers in normal canine eyes.

Tonometry, an indirect measurement of intraocular pressure (IOP), is important for the diagnosis and management of glaucoma and uveitis. The aim of this study was to compare the performance of three hand-held tonometers in normal canine eyes. Eyes from cadavers of dogs without observable ocular disease were used to compare tonometric measurements with direct manometry over a range of 7.4-65mmHg. In vivo measurements using the three tonometers in both eyes of 12 healthy dogs were compared. All tonometers significantly underestimated manometric values both ex vivo and in vivo. One tonometer showed a small fixed bias over the range of IOP, whilst the other two tonometers had a negative proportional bias. The results of this study show that differences exist between handheld tonometers across the clinically relevant range of IOP, and that all underestimate manometric measurements.

Schirmer tear test I in dogs: results comparing placement in the ventral vs. dorsal conjunctival fornix.

OBJECTIVE: To compare Schirmer tear test I (STT I) values obtained from placement of tear strips in the ventral and dorsal conjunctival fornices in dogs. PROCEDURE: Schirmer tear test I was performed on each eye of 16 clinically normal dogs (32 eyes) in a crossover study. Initial tear strip placement site was randomized for each eye. Alternative placement site measurements were obtained after 1 week. RESULTS: The mean (± standard deviation, SD) STT I for dorsal and ventral conjunctival fornices was 20.44 (±4.46) mm/min and 23.56 (±3.98) mm/min, respectively. STT I values obtained from the ventral conjunctival fornix were significantly greater than those obtained from the dorsal conjunctival fornix (P = 0.004). CONCLUSIONS: Schirmer tear test I values were significantly greater with tear strips placed in the ventral conjunctival fornix.

Development and Assessment of a Novel Canine Ex Vivo Corneal Model.

PURPOSE: To develop a novel ex vivo extended culture model of canine corneal epithelial cell wound healing. MATERIALS AND METHODS: Canine corneoscleral rims (CSR) were obtained and, after preparation for culture, were placed on a nutating scaffold and incubated in physiological conditions. In experiment 1, eight CSR in a serum-containing antimicrobial-fortified medium were monitored for epithelial integrity and bacterial infection up to 28 days in culture. CSR were assessed histologically at the end of the culture period end points 0, 7, 14, and 28 days with accompanying scanning electron microscopic (SEM) and transmission electron microscopic (TEM) evaluation. Samples for microbial culture were obtained at days 0, 3, 7, 14, and 28. In experiment 2, uniform 8-mm-diameter superficial corneal epithelial wounds were created and monitored for re-epithelialization in the same culture conditions or in a serum-free protein equivalent medium, with four CSR per group. Standardized digital images were obtained with cobalt filter at the time of fluorescein staining and media change every six hours. Image J imaging software was used to measure the area of fluorescein retention. Re-epithelialization rates were calculated and CSR then fixed for immunohistochemistry (IHC). RESULTS: All corneas survived to end points as described in experiment 1 with no evidence of contamination or compromised epithelial integrity. Histologically, a multilayered epithelium was maintained and corneal edema was not appreciated until day 14. SEM examination revealed epithelial cell layer confluence and migrating epithelial cells of normal cellular morphology with normal cell-cell interactions on TEM. In experiment 2, all eight corneas healed with a healing rate of 0.702 ± 0.130 mm2/h (1.25 mm/day epithelial cell migration rate) and were positive in IHC evaluation for markers of corneal fibrosis. CONCLUSION: This ex vivo canine corneal wound healing model is an appropriate and clinically relevant tool for assessment and modulation of epithelial wound healing.

Effect of topical ophthalmic latanoprost 0.005% solution alone and in combination with diclofenac 0.1% solution in healthy horses: a pilot study.

OBJECTIVE: To evaluate the effect of topical ophthalmic 0.005% latanoprost alone and in combination with 0.1% diclofenac on healthy horses. ANIMALS STUDIED: Twelve healthy adult horses. PROCEDURES: A randomized, masked crossover design was used with horses divided into three groups for once daily treatment in one randomly selected eye. For arm 1 of the study, Group D (n = 3) received 0.1% diclofenac, Group L (n = 3) received 0.005% latanoprost, and Group DL (n = 6) received 0.005% latanoprost and 0.2 ml of 0.1% diclofenac. For arm 2 of the study, horses from Group D and L were placed into Group DL and horses from Group DL were placed into either Group D or L. Evaluations of intraocular pressure (IOP), vertical pupil diameter, aqueous flare, conjunctival hyperemia, epiphora, blepharospasm, and blepharoedema were performed 4 times daily on days 1 and 2 (baseline), days 3 to 7 (arm 1 treatment), days 8 to 11 (washout), days 12 to 16 (arm 2 treatment), and days 17 and 18 (return to baseline). RESULTS: During the treatment period, significant reduction in IOP and vertical pupil diameter occurred in treated eyes of Groups L and DL, but not Group D. These variables did not differ significantly between Groups L and DL. Blepharospasm, blepharoedema, epiphora, and conjunctival hyperemia scores were significantly higher in Group L than in Groups D and DL. CONCLUSIONS: Latanoprost reduced IOP in healthy horses and signs of drug-induced discomfort were mitigated by concurrent use of diclofenac.

Effect of auriculopalpebral nerve block on Schirmer tear test I values in normal horses.

OBJECTIVE: To compare Schirmer tear test I (STTI) values collected in normal horses with and without an auriculopalpebral nerve block. PROCEDURE: Schirmer tear test I values were measured in 20 clinically normal horses (38 eyes) with a median age of 12 years. The order of eyes tested was randomized. Within 24-48 h, at the same time of day, tear measurements were collected again after administration of an auriculopalpebral nerve block. Each block was performed a minimum of 5 min prior to each STT I. A repeated-measures model was used to analyze differences between STT I values in eyes with and without nerve blocks incorporating within horse correlation between eyes. The mixed-model included fixed factors of treatment and eye and a random intercept for each horse. An unstructured covariance structure was used. RESULTS: On average, STT I values measured in eyes after auriculopalpebral nerve blocks were 0.55 mm/min greater than those without nerve blocks. This difference was not statistically significant (P = 0.5268). CONCLUSIONS: There was no effect of auriculopalpebral nerve block on STT I values in normal horses.

Hypokalemia and suspected renal tubular acidosis associated with topical carbonic anhydrase inhibitor therapy in a cat.

OBJECTIVE: To describe the occurrence of hypokalemia, metabolic acidosis, and suspected renal tubular acidosis associated with the administration of topical ophthalmic carbonic anhydrase inhibitor (CAI) in a cat. CASE SUMMARY: A 2-year-old, 5.3 kg, male, castrated, domestic short-haired cat developed hyporexia 6 weeks after starting topical ophthalmic dorzolamide 2% therapy for treatment of ocular hypertension. Two weeks later, the cat was evaluated for severe weakness, cervical ventroflexion, and anorexia. Plasma electrolyte and acid-base measurement revealed hypokalemia (K+ = 2.9 mmol/L; reference interval 3.8-5.4 mmol/L) and metabolic acidosis (plasma HCO3- = 9.8 mmol/L; reference interval 15-23 mmol/L) in the presence of a urine pH of 7.5 (reference interval 6.5-7.5). The pH abnormalities were consistent with a renal tubular acidosis. Clinical and biochemical abnormalities resolved with short-term supportive care, potassium supplementation, and discontinuation of dorzolamide therapy. NEW OR UNIQUE INFORMATION PROVIDED: This is the first report of hypokalemia and metabolic acidosis associated with topical CAI therapy in a cat.

Comparison of two- and three-times-daily topical ophthalmic application of 0.005% latanoprost solution in clinically normal dogs.

OBJECTIVE: To determine whether 2- or 3-times-daily application of topical ophthalmic 0.005% latanoprost solution is more effective at lowering intraocular pressure (IOP) in clinically normal dogs. ANIMALS: 9 clinically normal dogs. PROCEDURES: For each dog, I drop of latanoprost 0.005% solution was applied to 1 eye every 8 or 12 hours each day for 5 days; the contralateral eye received topical ophthalmic treatment with 1 drop of saline (0.9% NaCl) solution at the times of latanoprost application. Ocular examinations of both eyes were performed every 6 hours starting 48 hours prior to and ending 42 hours after the treatment period. Following a 5-week washout interval, the procedures were repeated but the previously latanoprost-treated eye of each dog received latanoprost application at the alternate frequency. RESULTS: Mean ± SD IOP reduction in the latanoprost-treated eyes was 31 ± 6.9% with 2-times-daily application and 33 ± 8.2% with 3-times-daily application. A 2-way repeated-measures ANOVA revealed significant differences in IOP with contributions by treatment (2 or 3 times daily), time of day (diurnal variation), and individual dog. The maximum mean daily IOP reduction in latanoprost-treated eyes was detected on day 3 of latanoprost treatment in each group. Eyes treated 3 times daily had significantly smaller pupil diameter and greater conjunctival hyperemia than eyes treated 2 times daily. CONCLUSIONS AND CLINICAL RELEVANCE: The clinical importance of the ocular hypotensive effects of 3-times-daily topical ophthalmic application of 0.005% latanoprost solution in dogs with glaucoma warrants investigation.

Schirmer tear test I and rebound tonometry findings in healthy calves.

OBJECTIVE: To describe Schirmer tear test I and rebound tonometry findings in healthy bovine calves. ANIMALS STUDIED: Thirty-three clinically normal dairy breed calves of mean (SD) 11 (1.1) weeks (range, 9.3-13.3 weeks) of age were evaluated. PROCEDURES: A Schirmer tear test I was performed on each eye followed by tonometry, using a TonoVet(®) without topical anesthesia. We report means (SD) and statistical analysis of data for each assay. RESULTS: For both Schirmer tear test (STT) I and tonometry, significant differences were not found between fellow eyes (P = 0.1536 and P = 0.83, respectively). The mean (SD) STT I value of all eyes was 20.4 (5.0) mm/min (range, 9-34 mm/min) while the mean (SD) intraocular pressure (IOP) value of all eyes was 15.2 (5.2) mmHg (range, 7-25 mmHg). CONCLUSIONS: This study reports normal data for the STT I and rebound tonometry in calves. This data may be useful in complete ophthalmic examinations of cattle, guiding diagnosis of glaucoma, uveitis, and keratoconjunctivitis sicca. However, results of these diagnostic modalities must be interpreted in light of clinical signs, given the wide range of normal values obtained in this study.

Assessment of oncolytic HSV efficacy following increased entry-receptor expression in malignant peripheral nerve sheath tumor cell lines.

Limited expression and distribution of nectin-1, the major herpes simplex virus (HSV) type-1 entry-receptor, within tumors has been proposed as an impediment to oncolytic HSV (oHSV) therapy. To determine whether resistance to oHSVs in malignant peripheral nerve sheath tumors (MPNSTs) was explained by this hypothesis, nectin-1 expression and oHSV viral yields were assessed in a panel of MPNST cell lines using γ134.5-attenuated (Δγ134.5) oHSVs and a γ134.5 wild-type (wt) virus for comparison. Although there was a correlation between nectin-1 levels and viral yields with the wt virus (R=0.75, P =0.03), there was no correlation for Δγ134.5 viruses (G207, R7020 or C101) and a modest trend for the second-generation oHSV C134 (R=0.62, P=0.10). Nectin-1 overexpression in resistant MPNST cell lines did not improve Δγ134.5 oHSV output. While multistep replication assays showed that nectin-1 overexpression improved Δγ134.5 oHSV cell-to-cell spread, it did not confer a sensitive phenotype to resistant cells. Finally, oHSV yields were not improved with increased nectin-1 in vivo. We conclude that nectin-1 expression is not the primary obstacle of productive infection for Δγ134.5 oHSVs in MPNST cell lines. In contrast, viruses that are competent in their ability to counter the antiviral response may derive benefit with higher nectin-1 expression.

Electroretinogram evaluation of equine eyes with extensive 'bullet-hole' fundic lesions.

OBJECTIVE: To evaluate the impact of extensive bullet-hole nontapetal fundic lesions in horses on retinal function as measured by full-field electroretinography (ERG). MATERIALS AND METHODS: Full-field ERG was performed on two horses with numerous bullet-hole lesions in the nontapetal fundus of both eyes. The ERG was first recorded from the eye with the more extensive lesions in response to a low-intensity light stimulus (0.03 cd s/m(2) ) that was given at times (T) T = 5, 10, 15, 20 min of dark adaptation. Consecutively, combined rod-cone response was evaluated bilaterally in response to high-intensity light stimulus (3 cd s/m(2) ), followed by cone function evaluation by flicker stimulus (3 cd s/m(2) at 30 Hz). Off-line analysis of the ERG recordings was then performed. RESULTS: Despite extensive bullet-hole lesions in the nontapetal fundus bilaterally in both horses, retinal function as measured by ERG did not show any observable deficits. The b-wave amplitude of the full-field ERG increased continuously from 5 to 20 min of dark adaptation peaking at 446 µv and 377 µv for horse number 1 and 2, respectively. The b-wave amplitudes of the combined rod-cone response were OS- 459 µv and OD- 392 µv for horse number 1 and OS- 491 µv and OD- 608 µv for horse number 2. The amplitude of the flicker ERG for horse number 1 was OS- 86 µv and OD- 110 µv and for horse number 2, OS- 80 µv and OD- 74 µv. CONCLUSIONS: Extensive bullet-hole chorioretinal lesions do not appear to compromise outer retinal function in these horses.