Corrigendum: Early-Onset Progressive Retinal Atrophy Associated with an IQCB1 Variant in African Black-Footed Cats (Felis nigripes).

This corrects the article DOI: 10.1038/srep43918.

Early-Onset Progressive Retinal Atrophy Associated with an IQCB1 Variant in African Black-Footed Cats (Felis nigripes).

African black-footed cats (Felis nigripes) are endangered wild felids. One male and full-sibling female African black-footed cat developed vision deficits and mydriasis as early as 3 months of age. The diagnosis of early-onset progressive retinal atrophy (PRA) was supported by reduced direct and consensual pupillary light reflexes, phenotypic presence of retinal degeneration, and a non-recordable electroretinogram with negligible amplitudes in both eyes. Whole genome sequencing, conducted on two unaffected parents and one affected offspring was compared to a variant database from 51 domestic cats and a Pallas cat, revealed 50 candidate variants that segregated concordantly with the PRA phenotype. Testing in additional affected cats confirmed that cats homozygous for a 2 base pair (bp) deletion within IQ calmodulin-binding motif-containing protein-1 (IQCB1), the gene that encodes for nephrocystin-5 (NPHP5), had vision loss. The variant segregated concordantly in other related individuals within the pedigree supporting the identification of a recessively inherited early-onset feline PRA. Analysis of the black-footed cat studbook suggests additional captive cats are at risk. Genetic testing for IQCB1 and avoidance of matings between carriers should be added to the species survival plan for captive management.

Efficacy and safety of suberoylanilide hydroxamic acid (Vorinostat) in the treatment of canine corneal fibrosis.

OBJECTIVE: Study aims were to evaluate the safety and efficacy of the Food and Drug Administration-approved drug Vorinostat [suberoylanilide hydroxamic acid (SAHA)] in the treatment of canine corneal fibrosis using an in vitro model. METHODS: Healthy donor canine corneas were collected and used to generate primary canine corneal fibroblasts (CCFs) by growing cultures in minimal essential medium supplemented with 10% fetal bovine serum. Canine corneal myofibroblasts, used as a model for corneal fibrosis, were produced by growing CCF cultures in serum-free medium containing transforming growth factor ß1 (1 ng/mL). Trypan blue exclusion assays were used to determine the optimal SAHA dose for this in vitro model. Four hour after culturing with TGFß1, CCF cultures were treated with 0.06% SAHA for 5 min (group 1) and for 24 h (group 2), representing single and multiple dose treatment regimes, respectively. Cultures were then further incubated in the presence of TGFß1 (1 ng/µL) under serum-free conditions until they reached 70% confluence. Trypan blue exclusion, immunocytochemistry, and TUNEL assays were used to evaluate the cytotoxicity of SAHA. Real-time PCR, western blot analysis, and immunocytochemistry were used to determine the efficacy of SAHA to inhibit canine corneal myofibroblast formation. RESULTS: Topical SAHA application in both treatment groups successfully decreased α-smooth muscle actin expression when compared to the TGFß1 only treatment group (P < 0.05). Tested SAHA did not affect CCF phenotype or cellular viability and did not cause significant cell death. CONCLUSIONS: Suberoylanilide hydroxamic acid safely and effectively inhibits TGFß1-induced CCFs transformation to myofibroblast in vitro.

Canine corneal fibroblast and myofibroblast transduction with AAV5.

OBJECTIVE: The aims of this study were (1) to determine the efficacy of adeno-associated vector serotype 5 (AAV5) for delivering gene therapy to canine corneal fibroblasts (CCFs) and myofibroblasts (CCMs) using enhanced green fluorescent protein (GFP) marker gene and (2) to evaluate the cytotoxicity of AAV5 to CCFs and CCMs using an in vitro model. METHODS: Healthy donor canine corneas were used to generate primary CCFs by growing cultures in minimal essential medium supplemented with 10% fetal bovine serum. Canine corneal myofibroblasts were produced by growing cultures in serum-free medium containing transforming growth factor ß1 (1 ng/mL). An AAV5 titer (6.5 × 10(12) µg/mL) expressing GFP under control of hybrid cytomegalovirus + chicken ß-actin promoters (AAV5-gfp) was used to transduce CCF and CCM cultures. Delivered gene expression in CCFs and CCMs was quantified using immunocytochemistry, fluorescent microscopy, and real-time PCR. Transduction efficacy of the AAV5 vector was determined by counting DAPI-stained nuclei and EGFP-positive cells in culture. Phase-contrast microscopy, trypan blue, and dUTP nick end labeling (TUNEL) assays were used to determine the toxicity and safety of AAV5 in this canine corneal model. RESULTS: Topical AAV5 application successfully transduced a significant population of CCFs (42.8%; P < 0.01) and CCMs (28%; P < 0.01). Tested AAV5 did not affect CCF or CCM phenotype or cellular viability and did not cause significant cell death. CONCLUSIONS: The tested AAV5 is an effective and safe vector for canine corneal gene therapy in this in vitro model. In vivo studies are warranted.

Comparison of ultrasonic Doppler flow monitor, oscillometric, and direct arterial blood pressure measurements in ill dogs.

OBJECTIVE: To compare blood pressure measurements obtained via ultrasonic Doppler flow monitor (DOP) and 2 oscillometric noninvasive blood pressure monitors (CAR and PAS) to invasive blood pressure (IBP) in hospitalized, conscious dogs with a range of blood pressures. DESIGN: Prospective clinical study. SETTING: University teaching hospital. ANIMALS: Eleven client-owned dogs aged between 4 months and 11.5 years (median 6 y), and weighing between 5.8 and 37.5 kg (median 30.2 kg). INTERVENTIONS: Blood pressure measurement. MEASUREMENTS AND MAIN RESULTS: Three consecutive measurements of systolic, diastolic, and mean arterial pressure (MAP) were recorded for each of the 3 indirect devices (only systolic for DOP), along with concurrent IBP measurements. The data were categorized into 3 groups: hypotensive (direct MAP<80 mm Hg), normotensive (80 mm Hgor=100 mm Hg), and hypertensive (direct MAP>100 mm Hg). Each indirect method was compared with the corresponding direct arterial pressure using the Bland-Altman method. Within the hypotensive group, each indirect method overestimated the corresponding IBP. Within the normotensive group all indirect systolic measurements and the PAS diastolic measurements underestimated the corresponding IBP. The remaining indirect measurements overestimated the corresponding IBP. Within the hypertensive group, DOP and CAR systolic measurements underestimated the corresponding IBP, and the remaining indirect measurements overestimated the corresponding IBP. In hypertensive dogs oscillometric systolic measurements were more accurate than MAP. In hypotensive dogs MAP measurements were more accurate than systolic measurements. All indirect measurements were most accurate in hypertensive dogs. CONCLUSIONS: The noninvasive blood pressure monitors in our study did not meet the validation standards set in human medicine. However, CAR diastolic and MAP measurements within the normotensive group, CAR MAP measurements within the hypertensive group, and PAS diastolic measurements in all groups were close to these standards. All indirect measurements showed greater bias during hypotension. Precision was poorer for all indirect systolic measurements than for MAP.