A systematic survey of loss-of-function variants in human protein-coding genes

Genome-sequencing studies indicate that all humans carry many genetic variants predicted to cause loss of function (LoF) of protein-coding genes, suggesting unexpected redundancy in the human genome. Here we apply stringent filters to 2951 putative LoF variants obtained from 185 human genomes to determine their true prevalence and properties. We estimate that human genomes typically contain ~100 genuine LoF variants with ~20 genes completely inactivated. We identify rare and likely deleterious LoF alleles, including 26 known and 21 predicted severe disease-causing variants, as well as common LoF variants in nonessential genes. We describe functional and evolutionary differences between LoF-tolerant and recessive disease genes and a method for using these differences to prioritize candidate genes found in clinical sequencing studies.     

An experimental assessment of landscape configuration effects on frog and toad abundance and diversity in tropical agro-savannah landscapes of southeastern Brazil

Amphibians are an imperiled group of vertebrate animals that typically have biphasic life histories involving a shift from aquatic larval habitats to terrestrial adult habitats. Habitat loss is the greatest threat to amphibians and the importance of the spatial configuration of terrestrial and breeding habitats upon the landscape in determining amphibian persistence is poorly known. The information gap is particularly acute in tropical landscapes that simultaneously host the greatest and most imperiled amphibian fauna on Earth. We installed 125 artificial ponds at different distances from forest fragments embedded in an agricultural matrix in southeastern Brazil. Constructed ponds attracted 13 anuran species; ponds at the forest fragment-matrix transition hosted a greater abundance and higher species richness of frogs and toads than those installed either far from or well within forest fragments. Forest fragments larger than 70 ha in agricultural areas harbored more individuals than smaller fragments. Our results indicate that landscape configuration has an important influence on frog and toad distribution and abundance in tropical agricultural landscapes and we suggest guidelines for maintaining favorable configurations of aquatic and terrestrial habitats for conserving this rich and imperiled species suite.     

Topographic placement of management practices in riparian zones to reduce water quality impacts from pastures

Riparian buffers, a best management practice (BMP), lessen environmental impacts caused by pasture-based agriculture by excluding livestock from streams and removing sediment and nutrients from overland flow before they enter the watercourse. Geospatial data analysis of digital elevation models (DEMs) can improve BMP placement by ensuring that BMPs intercept flow paths. Livestock heavy use areas and riparian buffers within 100?m of the stream were digitized using aerial photography of Spring Creek watershed (Pennsylvania, USA). Flow path lengths and stream entry points from heavy use areas, from all agricultural land uses, and from a random sample of points were calculated using 1, 10, and 30?m DEM resolutions. Euclidean distances identified the nearest stream entry point regardless of topography. Drainage areas for each streambank cell were evaluated using each DEM resolution. Topographic calculations differed significantly from Euclidean, with median differences of 14.3?m for flow path length and 24.1?m between stream entry points for the 10?m DEM. Existing buffers intercepted runoff from only 23?% of heavy use areas. Drainage areas ranged from one to hundreds of DEM cells. Any DEM is an arbitrary representation of a continuous surface; both resolution and processing method affect the suitability of such a representation for any given purpose. We found that 30?m DEMs did not provide reasonable flow path estimates at the scale of grazing agriculture in this region, while Pennsylvania 1?m DEMs were minimally smoothed during DEM preparation, resulting in erratic flow paths. The 10?m DEMs were the most suitable available DEM product, and should be used in conjunction with site visits for planning pasture BMP placement.     

Risk factors for feline infectious peritonitis in Australian cats

The objective of this study was to determine whether patient signalment (age, breed, sex and neuter status) is associated with naturally-occurring feline infectious peritonitis (FIP) in cats in Australia. A retrospective comparison of the signalment between cats with confirmed FIP and the general cat population was designed. The patient signalment of 382 FIP confirmed cases were compared with the Companion Animal Register of NSW and the general cat population of Sydney. Younger cats were significantly over-represented among FIP cases. Domestic crossbred, Persian and Himalayan cats were significantly under-represented in the FIP cohort, while several breeds were over-represented, including British Shorthair, Devon Rex and Abyssinian. A significantly higher proportion of male cats had FIP compared with female cats. This study provides further evidence that FIP is a disease primarily of young cats and that significant breed and sex predilections exist in Australia. This opens further avenues to investigate the role of genetic factors in FIP.     

Testicular fine needle aspiration cytology from a stallion with testicular degeneration after external genitalia trauma

A seven-year-old Quarter Horse had a serious external genitalia trauma with severe swelling of ventrum, penis, prepuce and scrotum after falling over a fence. Appropriated treatment was rapidly started after clinical examination. During recovery period, the spermatogenesis assess by semen evaluation was not possible due to stallion's inability to ejaculate. Therefore, for testicular evaluation fine needle aspiration cytology (FNAC) was performed. The first FNAC showed a deviation of germ cell line towards immature cells, mainly by primary spermatocytes (59.5%) with very few late spermatids and spermatozoa (2.5% each), and an increased Sertoli cells/germ cells ratio (478/100), which characterized testicular degeneration. One month after the first FNAC, the second exam presented a drastic decrease of Sertoli cells/germ cells ratio (7/100) and marked increase of mature cell number, specially by early and late spermatids (50% and 24.5%, respectively). In this case, the results of both FNAC could demonstrate a partial recovery of spermatogenesis activity. Two months later, the stallion had mated two mares successfully and they became pregnant. In conclusion, the adequate treatment allowed a complete recovery of the stallion's reproductive function, and since semen collection was impossible during treatment, testicular FNAC showed to be an efficient diagnostic method for evaluating acute damage in the spermatogenesis.     

Effects of the opioid remifentanil on the arrhythmogenicity of epinephrine in halothane-anesthetized dogs

Opioids may exert a protective effect against ventricular arrhythmias via a vagally mediated mechanism. This study evaluated the effects of the opioid remifentanil on arrhythmogenicity of epinephrine during halothane anesthesia. Eight dogs were assigned to 2 treatments in a randomized crossover design, with 1-week intervals between treatments. Anesthesia was maintained with 1.3% end-tidal halothane in oxygen and mechanical ventilation to maintain eucapnia. A constant rate infusion of remifentanil (0.72 microg/kg/min) was administered throughout the study in the experimental treatment, while control animals received physiologic saline as placebo. The arrhythmogenic dose of epinephrine (ADE), defined as 4 premature ventricular complexes (PVCs) within 15 s, was determined by administering progressively increasing infusion rates of epinephrine (2.5, 5.0, and 10 microg/kg/min), allowing 20 min intervals between each infusion rate. In both treatments, epinephrine infusions induced bradyarrhythmias and atrioventricular conduction disturbances, which were followed by escape beats and PVCs. In the remifentanil treatment, mean +/- s ADE values (11.3 +/- 4.9 microg/kg) did not differ from values observed in control animals (9.9 +/- 6.1 microg/kg). On the basis of the ADE model for assessing the arrhythmogenity of drugs during halothane anesthesia, the present study did not demonstrate a protective effect of remifentanil (0.72 microg/kg/min) against ventricular arrhythmias in dogs.     

Transiliac approach for exposure of lumbosacral intervertebral disk and foramen: technique description

Objective— To describe and evaluate a transiliac approach to the L7–S1 disk and intervertebral foramen in dogs.
Study Design— Cadaver study.
Animals— Fresh canine cadavers (n=10).
Methods— A craniolateral approach was made to each iliac wing of 10 fresh canine mixed breed cadavers. An 18 mm hole was drilled in a standardized position through the iliac wing. The musculature connected to the cranial aspects of the sacral wing was dissected and retracted cranially through this iliac window. Endoscopic exploration of the area was performed.
Results— The foramen and intervertebral disk were clearly observed in all specimens without iatrogenic injury of the L7 nerve branch. Access to the foramen was possible in 16 of 20 specimens without excision of the sacral wing; however, it was always partially excised to observe the intervertebral disk which lies more caudally and ventrally.
Conclusion— Transiliac approach to the lumbosacral joint allows direct exposure of the intervertebral disk and foramen through an iliac window. Endoscopic exploration provided good observation of the intervertebral disk and/or foramen.
Clinical Relevance— Transiliac approach could be used for lateral corpectomy and foraminotomy in dogs with degenerative lumbosacral stenosis caused by ventral or ventrolateral disk protrusion, foramen stenosis, or OCD of the L7–S1 joint. Clinical study will be necessary to evaluate the efficacy of this approach.     

Effects of 12 and 17 cmH2O positive end-expiratory pressure applied after alveolar recruitment maneuver on pulmonary gas exchange and compliance in isoflurane-anesthetized horses

Objective

To compare static compliance (C_st) and alveolar–arterial oxygen tension difference [P(a–a)O₂] between positive end-expiratory pressures (PEEP) of 7, 12 and 17 cmH₂O applied after an alveolar recruitment maneuver (RM) in isoflurane-anesthetized horses.